Comprehensive glycomics comparison between colon cancer cell cultures and tumours: implications for biomarker studies

Jenny H. L. Chik1, Jerry Zhou2, Edward S. X. Moh1, Richard Christopherson2, Stephen J. Clarke3, Mark P. Molloy1,4 and Nicolle H. Packer1

1Department of Chemistry and Biomolecular Sciences, Faculty of Science, Macquarie University, Sydney, Australia

2School of Molecular Bioscience, University of Sydney, Sydney, Australia. 3Department of Medicine, Royal North Shore Hospital, University of Sydney

4Australian Proteome Analysis Facility, Macquarie University, Sydney, Australia

Supplementary Figure 5 for

Suppl Figure 5. Annotated MS/MS for N-glycans. N-glycans have been numbered according to Supplementary Table 3. The 6 structures (number 119, 156, 157, 164, 165, 177) not confirmed by MS/MS are not shown. Glycan schemes were derived from GlycoWorkbench. Annotation was based on the presence of structural feature ions and common knowledge of known glycan synthetic pathways. Additional structural isomers cannot be excluded. Structural schemes not intended to show specific linkages (linkages not shown by linkage angles). Where ambiguity is indicated in parent structure, for example in the assignment of fucose, depiction of specific linkages and isomers are not intended for corresponding glycan fragments.

281.0308.2

323.1

350.2

383.1

407.0

425.0

463.1

508.1

527.4

553.2

570.2

600.3

635.3

671.2

689.3

731.3

774.3

803.8

835.4853.4

JC_SW620_NG_1_020312.D: -MS2(895.5), 41.3min #1593

0.0

0.5

1.0

1.5

4x10

Intens.

300 400 500 600 700 800 900 m/z

Glycan 1 Parent ion: m/z 895.5- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1

[M-H]- - C2H2O

287.3

305.0

323.1

340.9

383.3407.3

503.2

526.3

545.2

577.8

630.3

647.3

669.8

688.2

710.3

749.3

791.4

832.9

851.5

869.5

-MS2(911.4), 40.2min #1536

0

1000

2000

3000

4000

5000

6000

Intens.

200 300 400 500 600 700 800 900 m/z

Glycan 2 Parent ion: m/z 911.4- Composition: (Hex)3 (HexNAc)2

[M-H]- - C2H2O

323.0

365.1407.1

425.0

463.3

492.1

511.3

526.3

545.2

571.4

587.2

600.2

636.1

688.3

731.3

762.6

783.5 817.2

835.5

851.5

895.5

920.0

955.4

973.4

997.4

1015.5

1039.5

JC_SW1116_NG_1_020312.D: -MS2(1057.4), 47.2min #1782

0.0

0.5

1.0

1.5

2.0

2.5

3.0

4x10

Intens.

300 400 500 600 700 800 900 1000 1100 m/z

Glycan 3 Parent ion: m/z 1057.4- Composition: (Hex)3 (HexNAc)2 (Deoxyhexose)1

[M-H]- - C2H2O

425.1

485.1 575.2

629.2

708.3 827.3

869.4

911.5

1012.4

1073.4

1175.4

1404.6

-MS2(1235.6), 42.9min #1632

0

2

4

6

8

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 m/z

Glycan 4 Parent ion: m/z 1235.6- Composition: (Hex)2 + (Man)3(GlcNAc)2

221.1

323.0

435.1

536.1

617.2

668.3

707.3

869.4913.4

1031.5

1073.6

1174.4

1235.4

1277.5

-MS2(698.0), 36.8min #1408

0.0

0.5

1.0

1.5

2.0

5x10

Intens.

200 400 600 800 1000 1200 m/z

Glycan 5 Parent ion: m/z 698.32- Composition: (Hex)3 + (Man)3(GlcNAc)2

Notes: No distinction intended between six-branch and three-branch in fragmentation scheme.

204.2

323.1

408.3

535.7

575.2

617.2

647.3

711.3

792.0833.1

869.4911.3

1031.5

1073.1

1132.4

1174.6

1336.6

-MS2(698.4), 38.4min #1471

0

2000

4000

6000

8000

Intens.

200 400 600 800 1000 1200 1400 m/z

Glycan 6 Parent ion: m/z 698.32- Composition: (Hex)3 + (Man)3(GlcNAc)2

263.0

290.2

351.1

382.1

424.2

454.7

547.8

598.8

629.8

688.4

748.5874.2

911.4

994.5

1057.3

1183.3

1244.5

-MS2(710.8), 51.0min #1944

0

2000

4000

6000

8000

Intens.

200 400 600 800 1000 1200 m/z

Glycan 7 Parent ion: m/z 710.82- Composition: (Hex)1 (HexNAc)1 (Deoxyhexose)1 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arm and 6-arm in fragmentation scheme.

Glycan 8 Parent ion: m/z 718.82- Composition: (Hex)2 (HexNAc)1 + (Man)3(GlcNAc)2

221.1

321.2

425.2

491.3

535.7

587.2

637.8

689.2

754.3

828.5

869.4 910.2

971.31013.4

1073.61118.6

1158.6

1217.4

JC_SW1116_NG_1_020312.D: -MS2(718.8), 32.6min #1242

0

2000

4000

6000

Intens.

200 400 600 800 1000 1200 m/z

220.1

262.0

304.0

351.2

389.0

467.8

544.6

571.5

629.7

688.3

764.4

851.2

909.4

952.2

993.5

1059.4

1096.5

1261.6

1302.5

-MS2(731.3), 30.6min #1184

0

500

1000

1500

2000

Intens.

200 400 600 800 1000 1200 m/z

Glycan 9 Parent ion: m/z 731.32- Composition: (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2

220.0

262.1

304.1

350.1

424.3

508.1

571.2

650.8

679.3

754.4 810.9

951.5

981.1

1080.5

1163.7

1200.5

1243.6

1299.6 1363.6

-MS2(731.3), 40.4min #1560

0.00

0.25

0.50

0.75

1.00

1.25

4x10

Intens.

200 400 600 800 1000 1200 1400 m/z

Glycan 10 Parent ion: m/z 731.32- Composition: (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2

221.1

262.1

304.0

350.2

381.9446.2

549.3

629.7

710.2

795.1

891.4

952.5

994.1

1097.5

1202.3

1261.5

1300.7

1366.3

-MS2(731.4), 41.6min #1603

0.00

0.25

0.50

0.75

1.00

1.25

4x10

Intens.

200 400 600 800 1000 1200 m/z

Glycan 11 Parent ion: m/z 731.32- Composition: (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2

303.7

408.1

436.8

467.2

502.7

556.2

598.7

637.8

709.8

790.5

850.4

894.3

971.2

1053.3

1155.2

1198.6

1276.3

-MS2(739.3), 26.7min #1038

0

1000

2000

3000

4000

Intens.

400 600 800 1000 1200 m/z

Glycan 12 Parent ion: m/z 739.32- Composition: (Hex)1 (HexNAc)2 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arm and 6-arm in glycan fragment scheme.

262.0

304.1

366.9

405.9455.1491.2

526.6

556.2

658.3

728.2

810.6

873.4

910.7

993.5

1078.5

1299.6

1379.7

JC_SW1116_NG_2_020312.D: -MS2(739.4), 32.1min #1224

0

2000

4000

6000

Intens.

200 400 600 800 1000 1200 1400 m/z

Glycan 13 Parent ion: m/z 739.32- Composition: (Hex)1 (HexNAc)2 + (Man)3(GlcNAc)2

220.1

262.1

303.9

407.2

475.1

508.3

576.7

658.3

729.9

951.3

993.2 1055.61101.3

1156.5

1299.5

JC_SW620_NG_1_020312.D: -MS2(759.9), 27.5min #1065

0.0

0.5

1.0

1.5

2.0

4x10

Intens.

200 400 600 800 1000 1200 m/z

Glycan 14 Parent ion: m/z 759.92- Composition: (HexNAc)3 + (Man)3(GlcNAc)2

D-221

179.2262.8 323.1

374.1

485.2

515.2

617.3

668.3

698.3

749.4

791.4

869.3

929.2

1032.3 1193.6

1235.5

1336.4

-MS2(779.3), 35.7min #1359

0.0

0.5

1.0

1.5

2.0

2.5

5x10

Intens.

200 400 600 800 1000 1200 1400 m/z

Glycan 15 Parent ion: m/z 779.32- Composition: (Hex)4 + (Man)3(GlcNAc)2

Notes: Depiction of specific branching not intended in glycan fragment scheme.

323.2

347.1

383.3

425.0

503.2

596.2

668.2

698.3

758.4

869.1

911.5

1032.71075.7

1133.4

1193.5

1235.5

1336.6

JC_SW1116_NG_1_020312.D: -MS2(779.3), 36.9min #1413

0

1

2

3

4

5

4x10

Intens.

200 400 600 800 1000 1200 m/z

Glycan 16 Parent ion: m/z 779.32- Composition: (Hex)4 + (Man)3(GlcNAc)2

Notes: Depiction of specific branching not intended in glycan fragment scheme.

290.1383.4

600.2

655.2733.3

762.3

835.3

908.5

995.0 1057.8

1159.5

1216.6

1276.8

1345.6

1467.5

-MS2(783.3), 40.6min #1549

0

2000

4000

6000

Intens.

200 400 600 800 1000 1200 1400 m/z

Glycan 17 Parent ion: m/z 783.52- Composition: (Hex)1 (HexNAc)1 (NeuAc)1 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arm and 6-arm in glyan fragment scheme

290.1

323.3459.0

510.0

557.7

600.7

656.1

723.8

835.3

908.6 1097.5

1159.4

1216.4

1276.5

1345.6

1707.1

JC_SW837_NG_1_020312.D: -MS2(783.3), 48.2min #1858

0

2000

4000

6000

8000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Glycan 18 Parent ion: m/z 783.52- Composition: (Hex)1 (HexNAc)1 (NeuAc)1 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arm and 6-arm in glyan fragment scheme

222.1

290.1

323.1

438.7

519.2

600.3

672.8

715.3

754.3

836.4

907.4

961.3

998.3

1040.0

1114.4

1159.7

1217.5

1276.4

1345.6

1525.5

JC_SW837_NG_1_020312.D: -MS2(783.3), 55.7min #2128

0

1000

2000

3000

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 19 Parent ion: m/z 783.52- Composition: (Hex)1 (HexNAc)1 (NeuAc)1 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arm and 6-arm in glyan fragment scheme

341.2412.1

510.1

583.7629.8

673.2

710.4 772.3

809.1

911.5

1073.5

1240.4

1278.3 1406.6 1520.5

-MS2(791.8), 33.0min #1256

0.0

0.2

0.4

0.6

0.8

1.0

4x10

Intens.

400 600 800 1000 1200 1400 1600 m/z

Glycan 20 Parent ion: m/z 791.82- Composition: (Hex)2 (HexNAc)1 (Deoxyhexose)1 + (Man)3(GlcNAc)2

Notes: Specific linkage for fucose not intended in glycan scheme, i.e. fucose may be attached to galactose or GlcNAc.

244.7

281.0

364.1

424.2

468.0

508.3

553.1

609.2

709.8

771.3

851.5

923.4

962.5

1072.5

1132.4

1219.6

1337.8

1404.6

-MS2(791.8), 40.7min #1589

0

2000

4000

6000

Intens.

200 400 600 800 1000 1200 1400 m/z

Glycan 21 Parent ion: m/z 791.92- Composition: (Hex)2 (HexNAc)1 (Deoxyhexose)1 + (Man)3(GlcNAc)2

281.1323.1364.1

424.0

503.2

536.6

616.8

689.2

718.8

769.8

810.3

844.6

1014.4

1057.1

1235.4

1318.4

1377.5

1420.4

-MS2(799.8), 39.2min #1500

0.0

0.2

0.4

0.6

0.8

1.0

1.2

4x10

Intens.

200 400 600 800 1000 1200 1400 m/z

Glycan 22 Parent ion: m/z 799.82- Composition: (Hex)3 (HexNAc)1 + (Man)3(GlcNAc)2

323.0

408.2

485.1

536.3

595.1

629.2

711.3

787.2

911.21031.3

1073.5

1199.4

1240.5

1404.7

JC_SW480_3_NG_310712.D: -MS2(812.3), 36.2min #1370

0.0

0.2

0.4

0.6

0.8

1.0

4x10

Intens.

200 400 600 800 1000 1200 1400 m/z

Glycan 23 Parent ion: m/z 812.42- Composition: (Hex)1 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2

286.5

350.2

407.3

467.2

547.7

607.8

665.3

710.8

782.8

858.6 933.3

1020.31113.6

1258.5

1302.6

1363.5

1406.5

1461.8

-MS2(812.4), 31.9min #1230

0.0

0.2

0.4

0.6

0.8

1.0

4x10

Intens.

200 400 600 800 1000 1200 1400 m/z

Glycan 24 Parent ion: m/z 812.42- Composition: (Hex)1 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2

179.1

262.0

302.2

351.3

424.4

547.8

640.3 731.3

784.3

893.6

1016.5

1073.5

1156.5

1212.6

1257.3

1299.6

1404.6

1446.5

1522.5

-MS2(812.4), 33.1min #1275

0

1000

2000

3000

4000

Intens.

200 400 600 800 1000 1200 1400 m/z

Glycan 25 Parent ion: m/z 812.42- Composition: (Hex)1 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2

306.0

350.2

424.2

473.2

571.2

710.8

780.7

918.5

951.5

1080.6

1155.4 1259.6

1360.6

1404.5

1461.8 1560.6

-MS2(812.4), 43.3min #1647

0.0

0.2

0.4

0.6

0.8

1.0

4x10

Intens.

400 600 800 1000 1200 1400 1600 m/z

Glycan 26 Parent ion: m/z 812.42- Composition: (Hex)1 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2

262.1

303.1

407.2

485.1

537.2

637.3

719.3

760.4

790.3

892.4

1054.4

1115.4

1155.4

1193.5 1275.2

1418.4

1513.7

-MS2(820.3), 29.1min #1114

0.0

0.5

1.0

1.5

4x10

Intens.

200 400 600 800 1000 1200 1400 m/z

Glycan 27 Parent ion: m/z 820.32- Composition: (Hex)2 (HexNAc)2 + (Man)3(GlcNAc)2

262.0

304.1 364.0

467.2556.3

637.2

700.4

730.3

799.5

833.5

874.2 1072.21130.6

1199.5

1258.4

1317.5

1461.5

-MS2(820.4), 32.0min #1221

0

1

2

3

4

5

6

4x10

Intens.

200 400 600 800 1000 1200 1400 m/z

Glycan 28 Parent ion: m/z 820.32- Composition: (Hex)2 (HexNAc)2 + (Man)3(GlcNAc)2

[M-2H]2- - C2H2O

263.0

382.0

424.1

467.3

535.8

586.4

637.3

688.3730.4

799.3

911.2

1036.7

1096.5

1174.5

1276.4

1335.5 1419.6

1461.6

JC_SW837_NG_1_020312.D: -MS2(820.4), 40.5min #1551

0.00

0.25

0.50

0.75

1.00

1.25

1.50

4x10

Intens.

200 400 600 800 1000 1200 1400 m/z

Glycan 29 Parent ion: m/z 820.32- Composition: (Hex)2 (HexNAc)2 + (Man)3(GlcNAc)2

246.2

350.0

389.1

424.2

526.7

571.4 629.8

731.3

812.3891.3

951.31096.6

1154.4

1297.5

1463.6

1502.9

-MS2(832.9), 33.6min #1278

0.0

0.5

1.0

1.5

4x10

Intens.

200 400 600 800 1000 1200 1400 m/z

Glycan 30 Parent ion: m/z 832.92- Composition: (HexNAc)3 (Deoxyhexose)1 + (Man)3(GlcNAc)2

D-221

262.1

303.9

350.3

395.3

464.3

568.8

649.3

690.8 803.5

880.5951.2

1098.61160.5 1238.31298.5

1445.6

1568.7

-MS2(832.8), 39.9min #1524

0

1000

2000

3000

4000

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 31 Parent ion: m/z 832.92- Composition: (HexNAc)3 (Deoxyhexose)1 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arn and 6-arm in glycan fragmentation scheme.

262.0

302.2 350.2

389.3

424.1

466.8507.2

583.7

619.4

649.3

731.4

810.9 891.6 934.4

995.3

1100.6

1137.3 1188.8

1299.6

1362.4

1445.6

1586.5

JC_SW837_NG_1_020312.D: -MS2(832.8), 44.6min #1717

0

1000

2000

3000

4000

5000

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 32 Parent ion: m/z 832.92- Composition: (HexNAc)3 (Deoxyhexose)1 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arn and 6-arm in glycan fragmentation scheme.

350.3

528.2

571.0

627.4

751.8

803.8

875.1

1094.5

1154.6

1242.6

1297.5

1445.9

JC_SW1116_NG_2_020312.D: -MS2(832.8), 47.9min #1805

0

200

400

600

800

1000

1200

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 33 Parent ion: m/z 832.92- Composition: (HexNAc)3 (Deoxyhexose)1 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arn and 6-arm in glycan fragmentation scheme.

262.1

304.1

382.2

424.3

490.3

526.5

556.3

657.8

739.4

771.2

810.8

910.4952.51053.4

1113.4

1257.6

1317.6

1459.5

-MS2(840.9), 30.0min #1155

0

1

2

3

4

4x10

Intens.

200 400 600 800 1000 1200 1400 m/z

D-221

Glycan 34 Parent ion: m/z 840.92- Composition: (Hex)1 (HexNAc)3 + (Man)3(GlcNAc)2

Glycan 35 Parent ion: m/z 856.32- Composition: (Hex)1 (HexNAc)1 (Deoxyhexose)1 (NeuAc)1 +

(Man)3(GlcNAc)2

350.0

470.3529.5

600.3

693.8

774.3

835.4

893.5

960.3

1039.3

1159.5

1201.3

1261.5

1362.5

1550.5

1616.0

JC_TEPCAM+_NG_041111_BE4_01_5851.d: -MS2(856.5), 55.1min #1924

0

1000

2000

3000

4000

5000

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

290.2

346.2457.1

554.2

600.8

655.2

715.1

774.2

849.2

895.2

928.3

979.4

1054.3

1159.4

1201.5

1261.6

1303.5

1362.6

1423.6

1538.8

-MS2(856.4), 60.0min #2223

0

500

1000

1500

2000

2500

3000

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 36 Parent ion: m/z 856.32- Composition: (Hex)1 (HexNAc)1 (Deoxyhexose)1 (NeuAc)1 +

(Man)3(GlcNAc)2

Notes: Specific linkage of fucose not intended in glycan fragmentation scheme. I.e. fucose may be attached to outer-arm or chitobiose core.

Glycan 37 Parent ion: m/z 856.32- Composition: (Hex)1 (HexNAc)1 (Deoxyhexose)1 (NeuAc)1 +

(Man)3(GlcNAc)2

290.0

350.2

390.4 460.3

519.2

600.7

627.7

655.3693.7

774.4

835.3

909.4

997.81041.7

1117.3

1159.6

1262.4

1362.6

1422.9

1489.4

JC_T_EPCAM+_NG_020312.D: -MS2(856.3), 45.4min #1689

0.00

0.25

0.50

0.75

1.00

1.25

1.50

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Notes: No distinction intended between 3-arm/6-arm in glycan fragment scheme.

290.1 350.0

470.1

600.7

672.2

774.3

835.4

907.3

957.9

1032.9

1159.6

1261.5

1362.6

1429.3

1550.5

-MS2(856.3), 45.3min #1717

0

2000

4000

6000

8000

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 38 Parent ion: m/z 856.32- Composition: (Hex)1 (HexNAc)1 (Deoxyhexose)1 (NeuAc)1 +

(Man)3(GlcNAc)2

Glycan 39

263.1

323.2

434.2

485.3

596.4

677.4

779.4

850.4

1031.3

1193.5

1235.5

1355.6

1498.6

1560.7

-MS2(860.4), 47.8min #1674

0.0

0.5

1.0

1.5

2.0

2.5

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Parent ion: m/z 860.32-

Composition: (Hex)5 + (Man)3(GlcNAc)2 Notes: No distinction intended between 3-arm and 6-arm in glycan fragmentation scheme.

323.1

383.1

434.1

486.1

516.2

596.2

647.1678.2

749.4

779.4

830.3

911.3

1056.3

1150.5

1193.3

1398.5

1499.5

1640.6

-MS2(860.3), 35.9min #1371

0

1

2

3

4

5x10

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 40 Parent ion: m/z 860.32- Composition: (Hex)5 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arm and 6-arm in glycan fragmentation scheme.

323.1

383.4

485.3

545.2

597.3

677.8

749.2

830.4

906.0

1031.4

1080.9 1213.4 1337.41398.4

1499.6

1542.6 1663.8

-MS2(860.4), 44.1min #1674

0

1

2

3

4x10

Intens.

400 600 800 1000 1200 1400 1600 m/z

Glycan 41 Parent ion: m/z 860.42- Composition: (Hex)5 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arn and 6-arm in glycan fragmentation scheme.

305.2425.2

467.1

504.2

600.3

681.2

835.4

920.1

1040.6

1159.4

1219.5

1260.5

1321.3

1364.6

1506.4

1624.7

1732.3

-MS2(864.3), 39.0min #1492

0.0

0.2

0.4

0.6

0.8

1.0

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 42 Parent ion: m/z 864.42- Composition: (Hex)2 (HexNAc)1 (NeuAc)1 + (Man)3(GlcNAc)2

291.0

322.9

485.3

545.1

600.2

681.2

753.2

783.4

835.3

965.1

1003.6

1074.3

1110.4

1151.4

1191.5

1321.4

1439.6

1507.6

-MS2(864.4), 47.3min #1839

0

2000

4000

6000

Intens.

400 600 800 1000 1200 1400 m/z

Glycan 43 Parent ion: m/z 864.42- Composition: (Hex)2 (HexNAc)1 (NeuAc)1 + (Man)3(GlcNAc)2

308.1365.2

413.8

452.0

485.3

530.2

620.7

681.3

836.3

893.4

1057.7

1124.6

1219.7

1344.4

1387.6

1454.7 1517.7

JC_SW1116_NG_2_020312.D: -MS2(864.9), 38.9min #1488

0

2000

4000

6000

8000

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 44 Parent ion: m/z 864.92- Composition: (Hex)2 (HexNAc)1 (Deoxyhexose)2 + (Man)3(GlcNAc)2

Notes: Specific linkage of outer-arm fucose not intended in glycan fragmentation scheme.

305.2

364.1

510.2

617.2

690.2791.9

864.3

995.51073.5

1116.5

1235.5

1343.51402.6 1523.6

1689.6

-MS2(872.9), 37.2min #1425

0

2

4

6

8

4x10

Intens.

400 600 800 1000 1200 1400 1600 m/z

Glycan 45 Parent ion: m/z 872.92- Composition: (Hex)3 (HexNAc)1 (Deoxyhexose)1 + (Man)3(GlcNAc)2 Notes: Specific linkage of outer-arm fucose not intended in glycan fragmentation scheme.

247.1

323.1

366.0

409.2

485.1

608.7

647.3

689.8

761.8

851.8

932.5

1019.4

1236.5

1360.7

1421.6

1525.6

1587.5

JC_SW837_NG_1_020312.D: -MS2(872.8), 42.8min #1644

0

1

2

3

4

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 46 Parent ion: m/z 872.92- Composition: (Hex)3 (HexNAc)1 (Deoxyhexose)1 + (Man)3(GlcNAc)2

[M-2H]2- - C2H2O

305.2

350.1

426.1

485.3

575.9

608.3

688.8

726.3

782.8

852.3

892.4

1012.5

1055.3

1155.3

1234.6

1406.5

1525.8

1566.6

-MS2(872.9), 44.4min #1710

0

2000

4000

6000

8000

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 47 Parent ion: m/z 872.92- Composition: (Hex)3 (HexNAc)1 (Deoxyhexose)1 + (Man)3(GlcNAc)2

[M-2H]2- - C2H2O

349.9

409.2

588.2655.0

784.3

839.2

1057.5

1159.4

1220.5

1386.6

1445.6

1558.5

-MS2(885.4), 39.2min #1497

0

1000

2000

3000

4000

5000

Intens.

400 600 800 1000 1200 1400 1600 m/z

Glycan 48 Parent ion: m/z 885.42- Composition: (Hex)1 (HexNAc)2 (Deoxyhexose)2 + (Man)3(GlcNAc)2

[M-H]- -

350.2

407.3

510.2

571.2

701.8

804.4

875.7933.4

1080.4

1260.6

1427.7

1551.6

1607.8

1674.2

1913.6

-MS2(885.4), 40.9min #1560

0

1000

2000

3000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Glycan 49 Parent ion: m/z 885.42- Composition: (Hex)1 (HexNAc)2 (Deoxyhexose)2 + (Man)3(GlcNAc)2

Notes: Specific linkage of outer-arm fucose not intended in glycan fragmentation scheme.

368.0

396.3

445.0

511.4

549.1

630.8

692.3

722.7

773.2

803.9

868.7

1059.5

1099.3

1218.4

1260.6

1301.5

1368.4

1559.4

-MS2(885.4), 30.7min #1173

0

1000

2000

3000

4000

Intens.

400 600 800 1000 1200 1400 1600 m/z

Glycan 50 Parent ion: m/z 885.42- Composition: (Hex)1 (HexNAc)2 (Deoxyhexose)2 + (Man)3(GlcNAc)2

Notes: Distinction between 3-arm/6-arm and specific linkage of outer-arm fucose not intended in glycan fragmentation scheme.

[M-H]- -

262.1

330.0

467.2

556.9

638.2

710.3

812.9

1053.2

1180.5

1234.6

1276.5

1353.71413.7

1565.6

JC_SW1116_NG_1_020312.D: -MS2(893.3), 26.7min #1029

0

1000

2000

3000

4000

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 51 Parent ion: m/z 893.42- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arm and 6-arm in glycan fragment scheme.

262.0 391.2

527.2

629.2

710.4

812.3

910.41017.5

1097.6

1282.4

1384.4

1421.5

1565.7

JC_SW1116_NG_2_020312.D: -MS2(893.4), 30.6min #1170

0

500

1000

1500

2000

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 52 Parent ion: m/z 893.42- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arm and 6-arm in glycan fragment scheme.

350.2

386.3

425.6

509.2

571.3629.3 679.8

710.8

791.9

873.7

932.3 995.4

1077.31172.2

1217.6

1275.5

1363.41404.8

1567.9

-MS2(893.4), 33.9min #1300

0.00

0.25

0.50

0.75

1.00

1.25

4x10

Intens.

400 600 800 1000 1200 1400 1600 m/z

Glycan 53 Parent ion: m/z 893.42- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2

D-221

350.0407.1

468.2

509.2

557.3 619.8

712.4

803.8

872.5

923.3

995.11095.3

1137.4

1275.5

1318.51422.7

1565.7

1608.8

-MS2(893.4), 37.0min #1426

0

1

2

3

4

5

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 54 Parent ion: m/z 893.42- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2

307.2

409.1

486.3 570.3

710.3

863.9

1114.51278.71346.5

1404.5

1461.8

1565.5

1729.6

-MS2(893.4), 44.0min #1682

0.0

0.2

0.4

0.6

0.8

1.0

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Glycan 55 Parent ion: m/z 893.42- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arm and 6-arm in glycan fragment scheme.

262.8

350.1

424.2

465.4

547.6

640.8

710.9

811.3

872.9

914.4

951.2

1095.4

1277.5

1443.51608.6

-MS2(893.4), 45.2min #1740

0.0

0.2

0.4

0.6

0.8

1.0

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 56 Parent ion: m/z 893.42- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2

[M-2H]2- - C2H2O

287.1

350.3

407.1 527.3

619.4732.4

812.4

873.0

988.3

1095.6

1275.5

1366.2

1445.6 1608.3

1725.3

-MS2(893.4), 51.2min #1950

0.0

0.2

0.4

0.6

0.8

1.0

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Glycan 57 Parent ion: m/z 893.42- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2

247.3

350.2

424.3

525.6

711.3

812.4

873.3

1011.4

1062.4

1115.3

1275.5

1360.7

1444.7

1608.7

-MS2(893.4), 53.5min #2054

0

1000

2000

3000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Glycan 58 Parent ion: m/z 893.42- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2

276.1

424.2

485.1

527.4

629.3

718.8

820.8

871.4

933.6

1037.4

1216.5

1278.3

1360.5

1438.5

1580.6

1704.8

-MS2(901.3), 29.8min #1140

0.0

0.2

0.4

0.6

0.8

1.0

1.2

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Hex

Glycan 59 Parent ion: m/z 901.42- Composition: (Hex)3 (HexNAc)2 + (Man)3(GlcNAc)2

[M-2H]2- - Hex

[M-2H]2- - C2H4O2

Hex

Notes: Hex, Hexose

[M-H]- -

383.1

467.0

535.8

616.8

665.3

718.7

789.3

871.6

947.9

1002.1

1149.3

1215.5

1438.5

1504.5

1558.3

1606.7

1661.9

1702.7

1747.7

-MS2(901.4), 32.4min #1251

0

500

1000

1500

2000

Intens.

400 600 800 1000 1200 1400 1600 1800 m/z

Glycan 60 Parent ion: m/z 901.42- Composition: (Hex)3 (HexNAc)2 + (Man)3(GlcNAc)2

- 18

[M-2H]2- - Hex

Notes: Hex, Hexose; Fuc, Fucose

[M-2H]2- - C2H2O

304.1

424.2

467.1

550.1

637.3

718.3

820.3

871.3

952.3

1054.6

1169.91277.6

1317.7

1419.7 1623.6

1702.8

-MS2(901.3), 33.5min #1273

0.0

0.5

1.0

1.5

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 61 Parent ion: m/z 901.42- Composition: (Hex)3 (HexNAc)2 + (Man)3(GlcNAc)2

Hex

- H2O

[M-2H]2- - Hex

[M-2H]2- - C2H4O2

[M-H]- -

[M-H]- - Hex

Notes: Hex, Hexose; Fuc, Fucose

262.2

323.1

485.1

535.7

629.3718.8

811.3

871.4

936.2

1072.3

1235.5

1401.3

1475.8

1580.5

1624.8

-MS2(901.3), 34.8min #1323

0.0

0.5

1.0

1.5

2.0

2.5

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 62 Parent ion: m/z 901.42- Composition: (Hex)3 (HexNAc)2 + (Man)3(GlcNAc)2

[M-H]- - Hex

[M-H]- -

Hex

Notes: Hex, Hexose; Fuc, Fucose

[M-2H]2- - C2H4O2

[M-2H]2- - Hex

262.3364.1

425.2 491.2

527.2

571.8

611.2

658.3

730.7

816.3

884.3

1037.5

1115.5

1260.4

1317.6

1455.8

1608.5

-MS2(913.9), 28.0min #1074

0.0

0.5

1.0

1.5

2.0

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 63 Parent ion: m/z 913.92- Composition: (Hex)1 (HexNAc)3 (Deoxyhexose)1 + (Man)3(GlcNAc)2

[M-H]- -

Hex Fuc

D - 221

Notes: No distinction between 3-arm/6-arm intended in glycan fragment scheme. Hex, Hexose; Fuc, Fucose

262.0

304.3

350.3

508.2

598.1

657.8

729.2

832.9

893.8

952.7 1019.6

1114.6

1221.5

1316.4

1445.5

1607.8

1648.5

-MS2(913.8), 34.5min #1312

0.0

0.2

0.4

0.6

0.8

1.0

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 64

[M-2H]2- - Hex

[M-H]- -

Hex

Fuc

Parent ion: m/z 913.92-

Composition: (Hex)1 (HexNAc)3 (Deoxyhexose)1 + (Man)3(GlcNAc)2

Notes: Hex, Hexose; Fuc, Fucose

+

305.1

350.2

424.0

556.3

628.8

670.2

730.8

812.3

1036.4 1138.7

1256.4

1382.8

1463.8

1559.8

1607.5

1731.6

-MS2(913.9), 35.3min #1345

0.0

0.5

1.0

1.5

2.0

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 65

D-221

Notes: No distinction intended between 3-arm and 6-arm in glycan fragment scheme.

Parent ion: m/z 913.92-

Composition: (Hex)1 (HexNAc)3 (Deoxyhexose)1 + (Man)3(GlcNAc)2

382.1

424.2

472.1

525.6

670.2

738.9

820.3

913.5

1119.4

1238.7

1401.5

1480.7

1665.8

1721.5

-MS2(921.9), 31.8min #1210

0.00

0.25

0.50

0.75

1.00

1.25

1.50

4x10

Intens.

400 600 800 1000 1200 1400 1600 m/z

Glycan 66 Parent ion: m/z 921.92- Composition: (Hex)2 (HexNAc)3 + (Man)3(GlcNAc)2

D-221

2-

2-

Notes: No distinction intended between 3-arm and 6-arm in glycan fragment scheme.

262.2

308.2

350.2

389.1

529.4

571.3 641.8

678.8

751.8

832.9

904.4

1058.3

1155.5

1265.7

1359.5

1443.3

1502.5

1648.9

-MS2(934.4), 33.3min #1273

0

2000

4000

6000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

D-221

Glycan 67

Fuc

[M-2H]2- -

Fuc

Hex

Parent ion: m/z 934.32-

Composition: (HexNAc)4 (Deoxyhexose)1 + (Man)3(GlcNAc)2

[M-2H]2- - C2H4O2

[M-H]- -

Notes: No distinction intended for 3-arm/6-arm in glycan fragment scheme.

304.1

368.3

475.8

511.5

571.2

650.3

763.9

832.8904.3

951.4

1038.6

1096.5

1138.6

1302.5

1445.4

1501.5

1666.6

1769.8

-MS2(934.5), 43.3min #1689

0.0

0.2

0.4

0.6

0.8

1.0

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

4 x

Glycan 68 Parent ion: m/z 934.32- Composition: (HexNAc)4 (Deoxyhexose)1 + (Man)3(GlcNAc)2

290.2351.2 410.3

450.3

485.1

556.2

600.3

681.3

732.3

775.1

855.3

916.4981.4

1213.2

1321.6

1424.8

1524.7

1722.5

1789.7

-MS2(937.8), 43.9min #1666

0

2000

4000

6000

Intens.

400 600 800 1000 1200 1400 1600 1800 m/z

Glycan 69 Parent ion: m/z 937.32- Composition: (Hex)2 (HexNAc)1 (Deoxyhexose)1 (NeuAc)1 +

(Man)3(GlcNAc)2

290.1

350.1

425.0

536.4

600.2

681.7

775.3

835.3

916.3

1073.3

1217.6

1321.5

1420.41506.7

1585.6

1653.8

1772.6

-MS2(937.4), 51.4min #1959

0

1000

2000

3000

4000

5000

Intens.

400 600 800 1000 1200 1400 1600 1800 m/z

Glycan 70 Parent ion: m/z 937.32- Composition: (Hex)2 (HexNAc)1 (Deoxyhexose)1 (NeuAc)1 +

(Man)3(GlcNAc)2

323.1

383.0

468.2

515.2 597.2

634.2

677.4

705.3

758.3

809.3

860.8

911.9

971.41032.5

1152.2

1254.51477.7

1517.6

1561.5

1660.8

JC_SW480_3_NG_310712 recovered.D: -MS2(941.4), 34.7min #1316

0

2000

4000

6000

8000

Intens.

400 600 800 1000 1200 1400 1600 m/z

Glycan 71 Parent ion: m/z 941.42- Composition: (Hex)6 + (Man)3(GlcNAc)2

261.9340.7

383.1

435.7

469.1

515.2

596.8

647.1

677.6

758.2

809.4

860.3

971.2

1032.4

1116.7

1175.5

1336.6

1397.5

1499.5

1560.6

1661.8

1787.8

-MS2(941.4), 36.7min #1404

0.0

0.5

1.0

1.5

5x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Glycan 72 Parent ion: m/z 941.42- Composition: (Hex)6 + (Man)3(GlcNAc)2

262.3

323.1

407.2

485.2

545.1

596.7

677.7

758.3

809.4

860.8

911.3

971.31033.5 1174.4

1234.61294.5

1337.6 1499.6

1540.6

1660.5

1788.0

-MS2(941.4), 37.7min #1446

0

1

2

3

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Glycan 73 Parent ion: m/z 941.42- Composition: (Hex)6 + (Man)3(GlcNAc)2

Glycan 74 Parent ion: m/z 945.42- Composition: (Hex)3 (HexNAc)1 (NeuAc)1 + (Man)3(GlcNAc)2

306.3 365.1 407.2

467.1504.2

545.1 600.2

655.3

682.3

762.3

835.4

915.9

1040.3

1363.6 1483.6

1586.6

1668.5

-MS2(945.4), 42.4min #1611

0

1

2

3

4

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 75

290.0

374.3485.1

546.1

655.2683.4

762.3

835.4

915.4

1005.5

1054.8 1134.3

1175.5

1234.6

1303.6

1424.7

1466.7

1558.7

1601.7

1668.6

JC_SW837_NG_1_020312.D: -MS2(945.4), 49.9min #1917

0

2

4

6

8

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Parent ion: m/z 945.42-

Composition: (Hex)3 (HexNAc)1 (NeuAc)1 + (Man)3(GlcNAc)2

266.0 325.1

407.0492.3

716.3

762.8

819.3

876.9

1031.31115.5

1175.6

1235.5

1301.3

1372.5

1463.81526.6

1670.7

1715.8 1796.7

JC_SW1116_NG_1_020312.D: -MS2(945.9), 40.7min #1551

0.0

0.5

1.0

1.5

2.0

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 m/z

Glycan 76 Parent ion: m/z 945.92- Composition: (Hex)3 (HexNAc)1 (Deoxyhexose)2 + (Man)3(GlcNAc)2

366.1

412.2

446.7

485.1

575.0

665.3

698.3

737.4

770.8

852.3 925.41038.5

1235.6

1282.5

1397.5

1534.51686.7

JC_SW480_3_NG_310712.D: -MS2(953.9), 37.9min #1434

0

2000

4000

6000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

2-

Glycan 77 Parent ion: m/z 953.82- Composition: (Hex)4 (HexNAc)1 (Deoxyhexose)1 + (Man)3(GlcNAc)2

2-

323.1

383.0

465.2

507.2

688.3

770.8

851.7

995.3

1034.4

1086.5

1133.5

1216.5

1260.7

1339.5

1418.7

1523.6

1584.5

1689.7

1812.8

JC_SW837_NG_1_020312.D: -MS2(953.9), 40.0min #1534

0

2000

4000

6000

8000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Glycan 78 Parent ion: m/z 953.82- Composition: (Hex)4 (HexNAc)1 (Deoxyhexose)1 + (Man)3(GlcNAc)2

Notes: Specific linkage of outer-arm fucose not intended in fragment scheme. (i.e. fucose can be on Gal or GlcNac)

2-

2-

2-

327.2

368.2

424.1

519.3

600.3

656.2

701.8

753.4

835.3

876.4

927.9998.4

1099.41202.7

1321.4

1362.5

1464.5

1506.5

1548.5

1680.7

JC_SW620_NG_1_020312.D: -MS2(957.9), 46.3min #1779

0

1000

2000

3000

4000

5000

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Glycan 79 Parent ion: m/z 957.82- Composition: (Hex)1 (HexNAc)2 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2

Notes: No distinction intended in between 3-arm and 6-arm in glycan fragment scheme

290.1

368.2

407.3

529.2

571.2

630.6

679.0

758.3

835.3

1057.5

1182.5

1258.5

1302.2

1344.7

1389.6

1464.6

1515.51626.7

1697.8

1795.9

JC_SW620_NG_1_020312.D: -MS2(957.9), 54.6min #2068

0

1000

2000

3000

4000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Glycan 80 Parent ion: m/z 957.82- Composition: (Hex)1 (HexNAc)2 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2

Notes: No distinction intended in between 3-arm and 6-arm in glycan fragment scheme

Glycan 81 Parent ion: m/z 965.92- Composition: (Hex)2 (HexNAc)2 (NeuAc)1 + (Man)3(GlcNAc)2

Notes: No distinction intended in between 3-arm and 6-arm in glycan fragment scheme

300.2

424.3

453.1

526.2

569.1

600.2

655.3

693.3

782.8

835.3

884.8

945.4

1003.3

1057.5 1123.5

1201.4

1279.3

1362.5

1446.3

1524.4

1710.0

JC_SW480_3_NG_310712 recovered.D: -MS2(965.8), 47.1min #1805

0.0

0.5

1.0

1.5

2.0

2.5

5x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Glycan 82 Parent ion: m/z 965.92- Composition: (Hex)2 (HexNAc)2 (NeuAc)1 + (Man)3(GlcNAc)2

Notes: No distinction intended in between 3-arm and 6-arm in glycan fragment scheme

290.9

382.3

424.1

471.0537.3

674.4 753.4

782.8

835.4

885.4

935.9

1022.21063.5

1097.6

1159.6

1234.5

1275.4

1362.4

1479.5

1524.6

1641.7

1710.5

JC_SW480_3_NG_310712 recovered.D: -MS2(965.9), 54.8min #2093

0.0

0.5

1.0

1.5

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

309.1

350.1446.3

510.4

629.8

710.8783.4

885.4

945.9

1021.6

1095.3

1141.5

1218.4

1326.5

1405.5 1565.7

1607.7

1706.7

JC_SW1116_NG_1_020312.D: -MS2(966.4), 40.2min #1536

0

2000

4000

6000

8000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Notes: No distinction intended in between 3-arm and 6-arm in glycan fragment scheme

Glycan 83 Parent ion: m/z 966.52- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)2 + (Man)3(GlcNAc)2

[M-2H]2- - C2H2O

284.0

368.2

527.2

630.2

710.8779.9

885.5

1057.3

1262.6

1422.7

1530.7

1589.8

1754.9

1859.8

JC_TEPCAM+_NG_041111_BE4_01_5851.d: -MS2(966.5), 56.2min #1960

0

1000

2000

3000

4000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 2000 m/z

Parent ion: m/z 966.52-

Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)2 + (Man)3(GlcNAc)2 Glycan 84

Notes: Distinction intended in between 3-arm/6-arm and specific linkage of outer-arm fucose not intended in glycan fragment scheme

+

314.1

350.3

383.1

424.1

491.2

554.3

710.3

803.2

885.5

946.3

1013.4

1056.3

1095.5

1169.5

1225.3

1422.7

1463.6

1507.5

1564.6

1607.6

1709.6

1771.8

JC_SW837_NG_1_020312.D: -MS2(966.4), 48.0min #1851

0.00

0.25

0.50

0.75

1.00

1.25

1.50

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Parent ion: m/z 966.52-

Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)2 + (Man)3(GlcNAc)2 Glycan 85

Notes: No distinction intended in between 3-arm and 6-arm in glycan fragment scheme. Depiction of specific linkage of outer-arm fucose not intended.

[M-2H]2- - C2H2O

323.1

364.2

485.2

535.8

629.3

718.7

791.3

893.4

944.8

1047.5

1114.61217.5 1302.6

1378.8

1438.7

1546.51607.81726.7

-MS2(974.5), 28.9min #1105

0.0

0.5

1.0

1.5

4x10

Intens.

400 600 800 1000 1200 1400 1600 m/z

Parent ion: m/z 974.42-

Composition: (Hex)3 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2 Glycan 86

Notes: Depiction of specific linkage of outer-arm fucose not intended in fragment scheme (i.e. may be linked to Gal or GlcNAc)

D-221

364.1

467.2

510.0

637.8

718.8

791.4

893.5

944.4 1114.3

1260.5

1381.4

1439.51625.6

1728.7

1774.7

-MS2(974.4), 31.3min #1194

0

2000

4000

6000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Parent ion: m/z 974.42-

Composition: (Hex)3 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2 Glycan 87

Notes: Depiction of specific linkage of outer-arm fucose not intended in fragment scheme.

D-221

- H2O

307.2

368.2

424.2

485.2

563.4

629.1

718.2

802.4

892.4

944.4

1054.2

1216.7

1298.6

1439.8

1580.7

1771.7

1828.8

-MS2(974.4), 34.4min #1318

0.0

0.2

0.4

0.6

0.8

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Parent ion: m/z 974.42-

Composition: (Hex)3 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2 Glycan 88

D-221

290.8

350.1

424.2

467.2

510.2

637.3

718.5

761.9

811.9

893.4

1054.3

1155.4

1259.4

1332.6

1383.5

1437.7

1580.6

1772.6

-MS2(974.4), 38.0min #1458

0.0

0.2

0.4

0.6

0.8

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 m/z

Parent ion: m/z 974.42-

Composition: (Hex)3 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2 Glycan 89

368.2

407.2

504.2

637.3

710.9

802.4

884.8

953.5

1073.4

1173.3

1237.5

1437.6

1479.5

1566.6

1623.6

1769.6

JC_SW1116_NG_1_020312.D: -MS2(974.4), 39.3min #1501

0

2000

4000

6000

Intens.

400 600 800 1000 1200 1400 1600 1800 m/z

Parent ion: m/z 974.42-

Composition: (Hex)3 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2 Glycan 90

365.1

407.1

571.3650.3

731.2

804.8

885.9

956.9

1097.5 1260.5

1355.4

1464.7

1549.7

1605.6 1754.8

-MS2(986.9), 33.9min #1288

0.0

0.5

1.0

1.5

2.0

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Parent ion: m/z 986.92-

Composition: (Hex)1 (HexNAc)3 (Deoxyhexose)2 + (Man)3(GlcNAc)2 Glycan 91

Notes: No distinction intended between 3-arm and 6-arm in glycan representations. Depiction of specific linkage of outer-arm fucose not intended in fragment scheme.

D-221

350.1

407.4

466.7

629.8

730.8

773.8

885.8

966.9

1054.51117.6

1244.5

1337.4

1422.6

1559.7

1664.51753.7 1875.9

JC_SW837_NG_1_020312.D: -MS2(987.0), 43.5min #1672

0.00

0.25

0.50

0.75

1.00

1.25

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

Parent ion: m/z 986.92-

Composition: (Hex)1 (HexNAc)3 (Deoxyhexose)2 + (Man)3(GlcNAc)2 Glycan 92

Notes: No distinction intended between 3-arm and 6-arm in glycan representations.

382.1

424.1

485.1 572.0

637.8

739.3

816.3

914.4

964.9

1113.5

1258.5

1479.7

1627.7

1768.8 1932.0

-MS2(994.9), 30.0min #1146

0.00

0.25

0.50

0.75

1.00

1.25

1.50

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

Parent ion: m/z 994.92-

Composition: (Hex)2 (HexNAc)3 (Deoxyhexose)1 + (Man)3(GlcNAc)2 Glycan 93

Notes: Distinction of 3-arm/6-arm and specific linkage of outer-arm fucose not intended in glycan scheme (i.e. fucose can be linked to Gal or GlcNAc).

D-221

350.0

424.2

492.2

571.3

671.3

730.8

801.8

913.3

963.4

1113.6

1198.5

1299.61386.8

1481.6

1621.7

1754.0

1811.6

-MS2(994.9), 36.5min #1405

0.00

0.25

0.50

0.75

1.00

1.25

5x10

Intens.

400 600 800 1000 1200 1400 1600 1800 m/z

Parent ion: m/z 994.92-

Composition: (Hex)2 (HexNAc)3 (Deoxyhexose)1 + (Man)3(GlcNAc)2 Glycan 94

D-221

Notes: Distinction of 3-arm/6-arm and specific linkage of outer-arm fucose not intended in glycan scheme.

325.2

425.2

473.2

570.3

670.3

701.7

812.4

874.1

964.9

1079.5

1132.1

1294.4

1397.4

1438.6

1564.5

1607.4

1666.9

1768.8

1829.7

JC_SW837_NG_2_020312.D: -MS2(994.9), 38.1min #1458

0

1000

2000

3000

4000

5000

Intens.

400 600 800 1000 1200 1400 1600 1800 m/z

Notes: Distinction of 3-arm/6-arm and specific linkage of outer-arm fucose not intended in glycan scheme.

D-221

Parent ion: m/z 994.92-

Composition: (Hex)2 (HexNAc)3 (Deoxyhexose)1 + (Man)3(GlcNAc)2 Glycan 95

350.3

407.1

528.2

628.7

679.3

812.3913.8

965.5

1017.4

1078.5

1298.6

1427.4

1479.6

1648.6

1728.6

1810.9

1890.7

-MS2(994.9), 44.6min #1719

0.0

0.2

0.4

0.6

0.8

1.0

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

+

Parent ion: m/z 994.92-

Composition: (Hex)2 (HexNAc)3 (Deoxyhexose)1 + (Man)3(GlcNAc)2 Glycan 96

Notes: Distinction of 3-arm/6-arm not intended in glycan scheme.

381.8

424.2

466.3556.4

604.2

649.8

710.3

819.9

892.4

972.9

1098.5

1275.4

1437.7

1539.6

1605.51783.7

1827.0

1913.8

JC_SW480_4_NG_310712.D: -MS2(1002.7), 45.8min #1740

0.00

0.25

0.50

0.75

1.00

1.25

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 2000 m/z

Parent ion: m/z 1002.72-

Composition: (Hex)3 (HexNAc)3 + (Man)3(GlcNAc)2 Glycan 97

Notes: No distinction intended between 3-arm and 6-arm in glycan fragmentation scheme

350.3

465.2

571.2

612.3

733.3

799.8

907.0

992.41097.4

1158.7

1244.6

1301.4

1366.0

1463.6

1570.7

1631.7

1735.71796.9

1916.7

1993.8

JC_TEPCAM+_NG_041111_BE4_01_5851.d: -MS2(1007.5), 56.7min #1978

0

250

500

750

1000

1250

Intens.

200 400 600 800 1000 1200 1400 1600 1800 2000 m/z

Parent ion: m/z 1006.42-

Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)2 (Sulph)1 + (Man)3(GlcNAc)2 Glycan 98

[M-H]- SO3 H2O

H2O

H2O

H2O

290.0

470.3

545.3

652.3

835.9

981.3

1076.4

1134.6

1237.4

1338.8

1402.3

1483.4

1611.6

1746.5

1842.7

1938.7

-MS2(1018.4), 42.7min #1624

0

1000

2000

3000

4000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Parent ion: m/z 1018.42-

Composition: (Hex)3 (HexNAc)1 (Deoxyhexose)1 (NeuAc)1 +

(Man)3(GlcNAc)2

Glycan 99

Notes: Specific linkage of outer-arm fucose not intended in fragment scheme i.e. can be linked to Gal or GlcNAc

[M-2H]2- H2O

305.3

368.2

470.2

512.9

553.3

695.8

762.3

835.3

937.9

988.4

1183.6

1322.5

1401.6

1483.6

1533.6 1613.7

1669.5

1748.8

1812.7

1950.6

-MS2(1018.4), 46.8min #1767

0

1000

2000

3000

4000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 2000 m/z

Parent ion: m/z 1018.42-

Composition: (Hex)3 (HexNAc)1 (Deoxyhexose)1 (NeuAc)1 +

(Man)3(GlcNAc)2

Glycan 100

383.2

503.3545.2

596.5

668.4

758.3

839.3

941.3

992.3

1195.5

1314.6

1355.41481.6

1559.3

1722.3 1822.7

-MS2(1022.4), 38.5min #1476

0

2

4

6

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 m/z

Parent ion: m/z 1022.42-

Composition: (Hex)7 + (Man)3(GlcNAc)2

Glycan 101

290.1

383.0

467.1

593.1

655.3

762.8

835.4

1006.4

1216.3

1356.7 1514.8

1601.2

1645.5

1747.6

1830.6

JC_SW837_NG_1_020312.D: -MS2(1026.3), 39.8min #1524

0.0

0.5

1.0

1.5

2.0

2.5

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Parent ion: m/z 1026.42-

Composition: (Hex)4 (HexNAc)1 (NeuAc)1 + (Man)3(GlcNAc)2 Glycan 102

363.9 510.3

600.3

655.3

732.4

783.9

835.5

957.9

1008.9

1159.4 1242.6

1385.8

1568.6

1736.7

1873.6

JC_SW837_NG_1_020312.D: -MS2(1039.4), 41.8min #1602

0.0

0.5

1.0

1.5

2.0

2.5

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 2000 m/z

Parent ion: m/z 1038.92-

Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2 Glycan 103

Notes: Glycan 103 co-elutes with glycan 108. Distinction between 3-arm/6-arm and Specific linkage of outer-arm fucose not intended in glycan fragment scheme.

Parent ion: m/z 1038.92-

Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2 Glycan 104

364.0

409.2

470.1

510.2

570.3

628.9

655.3

702.4

770.4

855.8

885.8

1008.9

1115.3

1201.41257.6

1309.6

1386.4

1507.5

1568.91674.8

1752.7

1855.7

JC_SW837_NG_1_020312.D: -MS2(1038.9), 46.3min #1783

0.0

0.5

1.0

1.5

2.0

2.5

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

Notes: Distinction between 3-arm/6-arm and specific linkage of outer-arm fucose not intended in glycan fragment scheme.

351.2

381.8

471.3

526.3

574.3

600.3

655.3

702.2

754.4

782.8

835.4

875.9

956.9

1009.8

1054.4

1150.5

1203.5

1295.5

1344.4

1464.5

1526.6

1562.5

1608.8

1674.3

1709.7

1787.7

1901.7

JC_SW837_NG_1_020312.D: -MS2(1039.0), 47.7min #1836

0

2

4

6

8

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 m/z

Parent ion: m/z 1038.92-

Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2 Glycan 105

Notes: Distinction between 3-arm/6-arm not intended in glycan fragment scheme.

Parent ion: m/z 1038.92-

Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2 Glycan 106

408.2

471.0

528.3

572.2

600.3

655.2701.9

783.3

855.8

906.9

962.4 1019.4

1078.3

1159.7

1201.6

1277.4

1369.4

1423.8

1507.5

1568.6

1627.1

1670.6

1734.7

1788.6

1856.6

JC_SW837_NG_1_020312.D: -MS2(1039.0), 49.3min #1896

0

1000

2000

3000

4000

5000

6000

Intens.

400 600 800 1000 1200 1400 1600 1800 m/z

Notes: Distinction between 3-arm/6-arm and specific linkage of outer-arm fucose not intended in glycan fragment scheme.

Parent ion: m/z 1038.92-

Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2 Glycan 107

Notes: Distinction between 3-arm/6-arm and specific linkage of outer-arm fucose not intended in glycan fragment scheme.

290.0350.2

424.2

470.3

571.2

600.3

652.3

693.0

783.3

838.3

866.9

956.9

1018.5

1140.4

1200.8 1292.8

1387.3

1422.5

1524.6

1627.6 1728.7

1787.7

1831.0

1901.8

JC_SW837_NG_1_020312.D: -MS2(1038.9), 54.3min #2076

0.00

0.25

0.50

0.75

1.00

1.25

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 m/z

364.8

450.1 510.2

557.7

672.3

783.8

855.8

958.8

1009.0

1153.6

1276.5

1410.4

1568.5

1735.8

1874.5

-MS2(1039.4), 41.8min #1590

0.0

0.2

0.4

0.6

0.8

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

2x

Parent ion: m/z 1039.42-

Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2 Glycan 108

Notes: Specific linkage of outer-arm fucose not intended in glycan fragment scheme.

Parent ion: m/z 1039.42-

Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2 Glycan 109

323.0

359.2

425.9

526.2

605.3

637.3

689.3

763.4

802.7

877.5

936.3

991.6

1220.3

1448.5

1530.9

1586.5

1673.7

1754.6

JC_SW620_NG_1_020312.D: -MS2(1039.3), 46.0min #1767

0

1000

2000

3000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Notes: Distinction between 3-arm/6-arm and specific linkage of outer-arm fucose not intended in glycan fragment scheme.

Parent ion: m/z 1039.42-

Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2 Glycan 110

Notes: Specific linkage of outer-arm fucose not intended in glycan fragment scheme.

350.2

424.3

526.1

572.0

655.4

702.3

782.9

835.5

957.0

1018.0

1080.5 1204.5

1386.5

1482.5

1525.5

1570.61626.7

1709.7

JC_SW837_NG_1_020312.D: -MS2(1039.4), 47.4min #1825

0

2

4

6

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

2x

[M-2H]2- - C2H2O

Parent ion: m/z 1039.42-

Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2 Glycan 111

Notes: Specific linkage of outer-arm fucose not intended in glycan fragment scheme.

350.3

409.2

472.3

510.3553.3

637.3722.8

783.4

834.4

875.2

957.4

1019.0

1081.7

1182.4

1242.5

1363.6

1406.3

1568.7

1652.8

1710.7

1753.7

1856.71933.1

JC_SW837_NG_1_020312.D: -MS2(1039.4), 51.7min #1986

0.0

0.5

1.0

1.5

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

2x

[M-2H]2- - C2H2O

325.0

424.2

485.3

629.3

717.4818.2

864.9

945.9

1017.4

1235.6

1438.6

1516.5

1605.5

1727.7

1872.6

2097.0

JC_SW1116_NG_1_020312.D: -MS2(1047.4), 33.4min #1270

0

2000

4000

6000

8000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 2000 m/z

Parent ion: m/z 1047.72-

Composition: (Hex)3 (HexNAc)2 (Deoxyhexose)2 + (Man)3(GlcNAc)2 Glycan 112

D - 221

341.3

485.1

545.3

629.3

697.8

791.4

872.8

974.4

1026.0

1240.41320.5

1371.7

1601.6 1728.6

1888.7

2015.6

-MS2(1055.4), 34.2min #1300

0

2000

4000

6000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 2000 m/z

Parent ion: m/z 1055.42-

Composition: (Hex)4 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2 Glycan 113

369.0

427.8

485.1

629.3

688.8

799.8

883.4

965.9

1025.3

1133.61241.5

1342.5

1402.8

1585.8

1729.9

1884.6

1933.6

2053.9

-MS2(1055.3), 40.3min #1537

0

500

1000

1500

2000

2500

3000

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

Parent ion: m/z 1055.42-

Composition: (Hex)4 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2 Glycan 114

305.3

383.0

485.0

536.1

601.3

647.2 719.2

791.3

872.4

935.9

1025.3

1215.2

1317.6

1482.6

1583.6

1707.3

1841.7

2011.7

-MS2(1055.3), 32.1min #1225

0

1000

2000

3000

4000

5000

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

Parent ion: m/z 1055.42-

Composition: (Hex)4 (HexNAc)2 (Deoxyhexose)1 + (Man)3(GlcNAc)2 Glycan 115

+

Notes: Specific linkage of fucose not intended in glycan fragment scheme (i.e. may be outer-arm or core-fucose).

304.1

350.0

470.2

571.2

655.3

701.8

817.3

947.9

1029.9

1161.3

1344.8

1464.8

1565.6

1667.7

1750.6

1898.7

-MS2(1059.4), 38.5min #1489

0.0

0.2

0.4

0.6

0.8

1.0

1.2

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 m/z

D-221

Parent ion: m/z 1059.42-

Composition: (Hex)1 (HexNAc)3 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2 Glycan 116

Notes: No distinction intended between 3-arm/6-arm in glycan fragment scheme.

290.1

368.0

410.2

571.3644.2

718.3

835.4

876.4

961.4

1183.2

1344.6

1463.7

1547.6

1667.8

1750.7

1829.7

1899.7

-MS2(1059.4), 42.8min #1665

0

2000

4000

6000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

D-221

Parent ion: m/z 1059.42-

Composition: (Hex)1 (HexNAc)3 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2 Glycan 117

Notes: No distinction intended between 3-arm/6-arm in glycan fragment scheme.

350.2405.2

507.2

600.2 655.2

734.0

803.5

876.3

978.3

1038.3

1201.41262.6

1357.51428.6 1506.7

1565.6

1635.8 1751.7

1842.8

1898.9

JC_SW480_4_NG_310712.D: -MS2(1059.4), 51.9min #1983

0.0

0.2

0.4

0.6

0.8

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

Parent ion: m/z 1059.42-

Composition: (Hex)1 (HexNAc)3 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2 Glycan 118

Notes: No distinction intended between 3-arm/6-arm in glycan fragment scheme.

348.1

425.2570.3

641.4 723.3

812.4

884.9

987.0

1038.5

1115.3

1260.5

1426.7

1566.6

1626.6

1792.7

1915.7

-MS2(1068.0), 28.2min #1080

0.0

0.5

1.0

1.5

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 2000 m/z

D-221

Notes: No distinction intended between 3-arm/6-arm in glycan fragment scheme.

Parent ion: m/z 1068.02-

Composition: (Hex)2 (HexNAc)3 (Deoxyhexose)2 + (Man)3(GlcNAc)2 Glycan 120

+

368.2

424.1

479.0

629.3

698.8

812.4

884.8

987.0

1038.3

1113.41209.1

1261.51401.7

1625.6

1703.7

1774.6

1897.8

1957.9

-MS2(1068.0), 35.0min #1332

0

1

2

3

4

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 m/z

Parent ion: m/z 1068.02-

Composition: (Hex)2 (HexNAc)3 (Deoxyhexose)2 + (Man)3(GlcNAc)2 Glycan 121

D-221

+

Notes: No distinction intended between 3-arm/6-arm in glycan fragment scheme.

324.9

424.2

490.2

581.2

670.3

730.2

811.9

884.4

985.9

1037.9

1154.5

1225.6

1317.51464.7

1624.6

1728.6

1810.8

1915.9

-MS2(1068.0), 41.1min #1587

0.0

0.2

0.4

0.6

0.8

1.0

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 m/z

+

Glycan 122 Parent ion: m/z 1068.02- Composition: (Hex)2 (HexNAc)3 (Deoxyhexose)2 + (Man)3(GlcNAc)2

D-221

Notes: No distinction intended between 3-arm/6-arm in glycan fragment scheme.

308.2

350.2

424.2

512.3

599.8

739.3

782.3

820.0

872.4

994.9

1054.9

1140.5

1200.3

1298.41463.7

1640.5

1783.6

1866.6

1973.1 2056.2

-MS2(1075.9), 46.8min #1794

0.0

0.5

1.0

1.5

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 2000 m/z

Glycan 123 Parent ion: m/z 1075.92- Composition: (Hex)3 (HexNAc)3 (Deoxyhexose)1 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arm/6-arm in glycan fragment scheme.

424.1

508.1 571.1

722.8

799.4 913.0

956.1

1015.5

1162.6

1263.3

1360.5

1545.5

1608.3

1745.7

1874.3

1974.8

2045.8

-MS2(1075.9), 50.7min #1935

0

1000

2000

3000

4000

5000

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

Glycan 124 Parent ion: m/z 1075.92- Composition: (Hex)3 (HexNAc)3 (Deoxyhexose)1 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arm/6-arm in glycan fragment scheme.

368.3

466.1

544.2606.1

688.2

790.4

903.8

995.4

1054.9

1277.4

1363.6

1461.5

1607.7

1751.8

1866.7

1987.8

-MS2(1075.9), 53.7min #2037

0

2000

4000

6000

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

Glycan 125 Parent ion: m/z 1075.92- Composition: (Hex)3 (HexNAc)3 (Deoxyhexose)1 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arm/6-arm in glycan fragment scheme.

368.3

551.3

641.3

751.3

804.4

867.4

980.0

1058.4

1163.3 1240.5

1335.5

1445.5

1609.7

1746.7

1886.6

1942.0

2007.6

JC_SW480_3_NG_310712.D: -MS2(1080.5), 45.3min #1731

0.0

0.5

1.0

1.5

2.0

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

Glycan 126 Parent ion: m/z 1080.62- Composition: (HexNAc)4 (Deoxyhexose)3 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arm/6-arm in glycan fragment scheme.

310.2

350.2

424.2

553.5

657.8

739.3

840.4

933.4

1015.4

1066.5

1257.4 1339.5

1384.5

1460.5

1569.4

1662.9

1824.8

1972.0

2095.1

JC_SW620_NG_2_020312.D: -MS2(1096.5), 37.9min #1479

0.0

0.5

1.0

1.5

2.0

2.5

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 2000 m/z

+ 2 x

Glycan 127 Parent ion: m/z 1096.52- Composition: (Hex)2 (HexNAc)4 (Deoxyhexose)1 + (Man)3(GlcNAc)2

- H2O

Notes: No distinction intended between 3-arm/6-arm in glycan fragment scheme.

424.3

538.9

658.4

750.4

840.8

905.0

995.0

1066.5

1221.5

1521.5

1609.0

1681.6

1824.7

1953.6

2014.6

-MS2(1096.5), 39.8min #1539

0.0

0.5

1.0

1.5

2.0

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 2000 m/z

+ 2 x

Glycan 128 Parent ion: m/z 1096.52- Composition: (Hex)2 (HexNAc)4 (Deoxyhexose)1 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arm/6-arm in glycan fragment scheme.

341.1

485.3

545.2

647.2

707.3

749.6

809.2

920.8

1022.9

1073.9

1177.51356.6

1439.3

1559.7

1722.7

1841.6

1988.8

JC_SW480_3_NG_310712.D: -MS2(1103.4), 45.3min #1730

0

1

2

3

4

5

6

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

Glycan 129 Parent ion: m/z 1103.42- Composition: (Hex)8 + (Man)3(GlcNAc)2

Glycan 130 Parent ion: m/z 1111.42- Composition: (Hex)2 (HexNAc)2 (NeuAc)2 + (Man)3(GlcNAc)2

655.3 817.4 888.4

1001.0

1102.6

1405.8

1567.8

1641.9 1710.81773.0

1933.0

JC_TEPCAM+_NG_041111_BE4_01_5851.d: -MS2(1111.5), 53.6min #1872

0.0

0.5

1.0

1.5

2.0

2.5

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

Notes: No distinction intended between 3-arm/6-arm in glycan fragment scheme.

[M-H]- -

[M-2H]2- - H2O

346.1510.2

655.3

782.8835.3

965.9

1102.5

1386.6

1479.7

1567.81641.7

1712.6

1770.7

1933.8

2123.8

JC_SW480_3_NG_310712 recovered.D: -MS2(1111.4), 57.4min #2186

0.0

0.2

0.4

0.6

0.8

1.0

1.2

5x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 2200 m/z

Glycan 131 Parent ion: m/z 1111.42- Composition: (Hex)2 (HexNAc)2 (NeuAc)2 + (Man)3(GlcNAc)2

Notes: No distinction intended between 3-arm/6-arm in glycan fragment scheme.

[M-H]- -

[M-2H]2- - H2O

350.3

408.1 510.3

553.3

600.2

655.3

774.9

835.4

939.9

1030.0

1091.5

1201.6

1261.4

1368.4

1570.6

1672.51855.8

-MS2(1112.0), 46.3min #1752

0.0

0.5

1.0

1.5

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

+

Notes: Distinction between 3-arm/6-arm and specific linkage of outer-arm fucose not intended glycan fragment scheme..

Glycan 132 Parent ion: m/z 1111.92- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)2 (NeuAc)1 + (Man)3(GlcNAc)2

+

332.2

409.1 510.2

570.5

655.3

835.4

949.4

1029.9

1091.0

1202.41274.3

1346.4 1447.6

1568.6 1670.71855.8

1899.8

JC_SW837_NG_2_020312.D: -MS2(1112.0), 50.7min #1925

0

1

2

3

4

5

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 m/z

Notes: Distinction between 3-arm/6-arm and specific linkage of outer-arm fucose not intended glycan fragment scheme..

Glycan 133 Parent ion: m/z 1111.92- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)2 (NeuAc)1 + (Man)3(GlcNAc)2

350.0

409.1510.2

571.3

657.2 708.7

774.4

856.5

940.0

979.4 1031.3

1102.5

1160.41261.6

1344.51422.8

1549.7

1670.7 1770.8

1881.8

1932.7

JC_SW837_NG_1_020312.D: -MS2(1112.0), 53.2min #2037

0

1

2

3

4

5

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

+

Glycan 134 Parent ion: m/z 1111.92- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)2 (NeuAc)1 + (Man)3(GlcNAc)2

[M-H]- -

[M-2H]2- - H2O

Notes: Distinction between 3-arm/6-arm and specific linkage of outer-arm fucose not intended glycan fragment scheme..

325.4470.2

512.1

571.3 656.1

731.2774.7

817.4

856.4

943.4

979.4

1029.8

1091.0

1240.7

1344.6

1423.8

1491.7

1608.6

1670.7

1716.6

1772.7

1933.7

JC_SW837_NG_1_020312.D: -MS2(1111.9), 55.5min #2119

0

1000

2000

3000

4000

5000

6000

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

+

Glycan 135 Parent ion: m/z 1111.92- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)2 (NeuAc)1 + (Man)3(GlcNAc)2

[M-H]- -

Notes: Distinction between 3-arm/6-arm and specific linkage of outer-arm fucose not intended glycan fragment scheme..

[M-2H]2- - H2O

[M-2H]2- - C2H2O

Glycan 136 Parent ion: m/z 1111.92- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)2 (NeuAc)1 + (Man)3(GlcNAc)2

350.2

410.1

511.4

571.1

655.3

702.3

855.9

924.1

965.2

1029.91102.9

1200.5

1245.31316.5

1422.5

1479.7

1549.6

1672.6 1754.7

1855.8

1932.8

1979.7 2189.8

JC_SW837_NG_1_020312.D: -MS2(1112.0), 56.6min #2158

0.0

0.2

0.4

0.6

0.8

1.0

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 2200 m/z

Notes: Distinction between 3-arm/6-arm and specific linkage of outer-arm fucose not intended glycan fragment scheme..

[M-H]- -

[M-2H]2- - H2O

+

350.0 438.2

493.2

551.2

620.8

655.1

714.2

774.8

856.3

925.8

1050.5

1104.7

1159.3

1242.4

1299.5

1346.6

1409.6

1460.7

1585.8

1653.6

1695.7

1815.7

1880.7

1973.7 2045.8

JC_SW480_3_NG_310712 recovered.D: -MS2(1133.0), 48.8min #1871

0

2000

4000

6000

8000

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

Glycan 137 Parent ion: m/z 1132.42- Composition: (Hex)1 (HexNAc)3 (Deoxyhexose)2 (NeuAc)1 + (Man)3(GlcNAc)2

Notes: Distinction between 3-arm/6-arm and specific linkage of outer-arm fucose not intended glycan fragment scheme. Fuc, Fucose.

[M-H]- -

[M-2H]2- - Fuc

410.2

452.1

533.2

572.5

655.3

716.4

835.4

884.9

978.0

1059.5

1119.9

1200.6

1307.5

1371.6

1464.1

1548.7

1661.2

1729.01770.6

1829.7

1913.8

1990.7 2062.9

2109.0

JC_TEPCAM+_NG_041111_BE4_01_5851.d: -MS2(1140.5), 51.3min #1794

0

500

1000

1500

2000

2500

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 2200m/z

Glycan 138 Parent ion: m/z 1140.42- Composition: (Hex)2 (HexNAc)3 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2

Notes: Distinction between 3-arm/6-arm not intended glycan fragment scheme. Hex, hexose.

+

[M-2H]2- - Hex

[M-2H]2- - C2H2O

[M-H]- -

323.0

390.3

574.2

692.7

835.2

885.9

958.0

1059.5

1111.5

1209.4

1404.7

1487.6

1628.7

1753.7

1878.6

1938.0

2058.7

JC_SW837_NG_2_020312.D: -MS2(1140.9), 34.3min #1304

0

1000

2000

3000

4000

5000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 2000 m/z

Glycan 139 Parent ion: m/z 1140.42- Composition: (Hex)2 (HexNAc)3 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2

Notes: Distinction between 3-arm/6-arm and specific linkage of outer-arm fucose not intended glycan fragment scheme. Fuc, Fucose.

D - 221

350.2

407.2

470.2

655.3

715.3

817.4

885.3

968.6

1058.5

1119.5

1225.4

1404.61548.7

1727.7

1912.8

2102.8 2193.8

JC_SW620_NG_2_020312.D: -MS2(1140.4), 39.8min #1553

0

2

4

6

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 2000 2200 m/z

Glycan 140 Parent ion: m/z 1140.42- Composition: (Hex)2 (HexNAc)3 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2

Notes: Distinction between 3-arm/6-arm not intended glycan fragment scheme.

D - 221

350.2426.2

471.0

571.3670.4

835.3

884.4

961.4

1058.4

1110.5

1285.3

1549.6

1626.7

1728.6

1828.9

1916.5

1990.8

2036.2

JC_SW837_NG_1_020312.D: -MS2(1140.4), 44.5min #1713

0.0

0.5

1.0

1.5

2.0

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

D - 221

Glycan 141 Parent ion: m/z 1140.42- Composition: (Hex)2 (HexNAc)3 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2

Notes: Distinction between 3-arm/6-arm not intended glycan fragment scheme.

Glycan 142 Parent ion: m/z 1140.42- Composition: (Hex)2 (HexNAc)3 (Deoxyhexose)1 (NeuAc)1 + (Man)3(GlcNAc)2

Notes: Distinction between 3-arm/6-arm not intended glycan fragment scheme.

382.1

424.2

536.3

571.4655.3

712.2

753.3

782.8

835.4884.3

961.4

1059.4

1111.0

1225.6

1277.61345.6

1464.8

1547.5 1626.8

1728.7

1831.7

1930.8

1990.7

2060.6

JC_SW837_NG_1_020312.D: -MS2(1140.4), 44.5min #1711

0.0

0.5

1.0

1.5

2.0

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

D - 221

350.3

453.4

571.3

677.5

722.8

804.4

885.4

954.0

1060.0

1098.61180.5

1260.6

1386.5

1427.6

1478.5

1609.7

1771.8

1861.71939.7

JC_SW1116_NG_1_020312.D: -MS2(1141.0), 33.8min #1287

0

2

4

6

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

Glycan 143 Parent ion: m/z 1141.02- Composition: (Hex)2 (HexNAc)3 (Deoxyhexose)3 + (Man)3(GlcNAc)2

Notes: Distinction between 3-arm/6-arm not intended glycan fragment scheme.

D - 221

350.1

424.3

510.2

652.3

716.3

835.4

885.4

947.3

1059.0

1120.1

1242.5

1334.6

1457.7

1626.8

1770.7

1915.7

2004.6

-MS2(1141.0), 39.6min #1512

0.0

0.5

1.0

1.5

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 2000 m/z

Glycan 144 Parent ion: m/z 1141.02- Composition: (Hex)2 (HexNAc)3 (Deoxyhexose)3 + (Man)3(GlcNAc)2

D - 221

Notes: Distinction between 3-arm/6-arm not intended in glycan fragment scheme.

325.1

409.2

510.3

570.3

632.2 722.9

798.3

884.8

978.4

1059.0

1120.5

1223.5

1390.5

1445.7

1591.6

1771.81913.9

2064.8

-MS2(1141.3), 49.8min #1915

0.0

0.2

0.4

0.6

0.8

1.0

1.2

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

SW837 a

Glycan 145 Parent ion: m/z 1141.02- Composition: (Hex)2 (HexNAc)3 (Deoxyhexose)3 + (Man)3(GlcNAc)2

Notes: Distinction between 3-arm/6-arm and specific linkage of outer-arm fucose not intended in glycan fragment scheme.

D - 221

365.2

424.2

490.1

655.2

702.3

835.3

884.5

965.4

1067.5

1118.9

1241.7 1316.31387.2

1471.4 1566.51641.6

1752.81889.5

2006.7

2093.8

JC_SW480_3_NG_310712 recovered.D: -MS2(1148.5), 48.5min #1857

0

1

2

3

4

5

6

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

Glycan 146 Parent ion: m/z 1141.02- Composition: (Hex)2 (HexNAc)3 (Deoxyhexose)3 + (Man)3(GlcNAc)2

Notes: Distinction between 3-arm/6-arm branching not intended in glycan fragment scheme. Depiction of specific branching also not intended.

365.1 424.0 532.2706.2

795.3

835.4

875.3

965.4

1067.4

1118.4

1318.5 1423.8

1566.7

1608.71737.7

1890.9

2009.9

JC_SW480_4_NG_310712.D: -MS2(1148.4), 54.7min #2082

0.00

0.25

0.50

0.75

1.00

1.25

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

Glycan 147 Parent ion: m/z 1148.42- Composition: (Hex)3 (HexNAc)3 (NeuAc)1 + (Man)3(GlcNAc)2

Notes: Distinction between 3-arm/6-arm branching not intended in glycan fragment scheme. Depiction of specific branching also not intended.

424.2

465.5

570.3

673.5

711.5

818.9

892.9

966.41067.4

1128.5

1200.4

1261.5

1378.7

1461.7

1640.6

1727.7

1873.8

1929.9

1974.8

2093.5

JC_SW837_NG_1_020312.D: -MS2(1148.9), 53.1min #2035

0.0

0.2

0.4

0.6

0.8

1.0

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

Glycan 148 Parent ion: m/z 1149.12- Composition: (Hex)3 (HexNAc)3 (Deoxyhexose)2 + (Man)3(GlcNAc)2

Notes: Distinction between 3-arm/6-arm branching not intended in glycan fragment scheme. Depiction of specific branching also not intended.

[M-2H]2- - C2H2O

325.2

364.2

424.1

510.4570.2

679.3

812.4

894.9

966.4

1019.1

1119.0

1200.81276.7

1326.4

1383.4

1461.7

1579.6

1626.81709.6

1787.6

1852.8

1972.8

2076.8

JC_SW837_NG_1_020312.D: -MS2(1148.9), 59.5min #2259

0

2000

4000

6000

8000

Intens.

200 400 600 800 1000 1200 1400 1600 1800 2000 2200 m/z

Glycan 149 Parent ion: m/z 1149.12- Composition: (Hex)3 (HexNAc)3 (Deoxyhexose)2 + (Man)3(GlcNAc)2

Notes: Distinction between 3-arm/6-arm branching and specific linkage for fucose not intended in glycan fragment scheme. Depiction of specific branching also not intended.

[M-2H]2- - C2H2O

381.1

424.0

466.2

570.2

647.2701.7

773.8

884.7

966.0

1118.5

1200.4

1275.71403.8

1480.01623.5

1678.4 1771.9

1844.7

1890.7

1933.8

2007.9

2095.7

JC_SW480_3_NG_310712 recovered.D: -MS2(1148.9), 56.4min #2148

0

1000

2000

3000

4000

5000

6000

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

+ 2x

Glycan 150 Parent ion: m/z 1149.12- Composition: (Hex)3 (HexNAc)3 (Deoxyhexose)2 + (Man)3(GlcNAc)2

Notes: Distinction between 3-arm/6-arm and specific linkage for outer-arm fucose not intended in glycan fragment scheme. Depiction of specific branching also not intended.

405.0

472.1613.3

756.3

794.8

877.4

960.3

1071.4

1200.4 1411.5 1610.7

1754.8

1922.8

2016.82080.9

JC_SW480_4_NG_310712.D: -MS2(1153.0), 49.4min #1881

0

1

2

3

4x10

Intens.

200 400 600 800 1000 1200 1400 1600 1800 2000 m/z

Glycan 151 Parent ion: m/z 1152.92- Composition: (HexNAc)4 (Deoxyhexose)2 (NeuAc)1 + (Man)3(GlcNAc)2

Notes: Distinction between 3-arm/6-arm and specific linkage for outer-arm fucose not intended in glycan fragment scheme.

425.1

571.4

670.3 740.3

803.0

921.9

994.9

1096.5

1148.0

1463.6

1593.3

1810.8

1972.8 2177.3

-MS2(1177.5), 39.1min #1495

0.0

0.5

1.0

1.5

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

Glycan 152 Parent ion: m/z 1177.52- Composition: (Hex)3 (HexNAc)4 (Deoxyhexose)1 + (Man)3(GlcNAc)2

Notes: Distinction between 3-arm/6-arm and depiction of specific branching not intended in glycan fragment scheme.

+

388.8

452.2 509.9

571.4

655.3774.4

857.0

929.3

982.4

1103.5

1164.5

1261.5

1345.6

1418.4

1550.7

1714.7

1817.7

1881.7

2020.8

2079.9

JC_SW837_NG_1_020312.D: -MS2(1184.4), 49.6min #1906

0.0

0.5

1.0

1.5

2.0

2.5

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 2200 m/z

Glycan 153 Parent ion: m/z 1184.52- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 (NeuAc)2 + (Man)3(GlcNAc)2

[M-H]- -

[M-2H]2- - C2H2O

- H2O

Notes: Distinction between 3-arm/6-arm not intended in glycan fragment scheme.

386.1 526.1 655.3

783.0

817.3 958.4

1039.0

1175.9

1260.7 1363.5 1557.6

1691.6

1787.7 1917.0

2000.8

2078.8

2153.8

JC_SW837_NG_1_020312.D: -MS2(1184.4), 56.7min #2163

0

2

4

6

8

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 2200 m/z

Glycan 154 Parent ion: m/z 1184.52- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 (NeuAc)2 + (Man)3(GlcNAc)2

[M-H]- -

[M-2H]2- - H2O

- H2O

Notes: Distinction between 3-arm/6-arm not intended in glycan fragment scheme.

571.3 638.4

856.9

919.4

1039.5

1176.0

1446.6 1507.4

1625.8

1714.6

1787.9

1919.7

2001.9

2078.9

JC_SW837_NG_1_020312.D: -MS2(1184.5), 60.9min #2307

0

2

4

6

8

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

Glycan 155 Parent ion: m/z 1184.52- Composition: (Hex)2 (HexNAc)2 (Deoxyhexose)1 (NeuAc)2 + (Man)3(GlcNAc)2

[M-H]- -

[M-2H]2- - H2O

Notes: Distinction between 3-arm/6-arm not intended in glycan fragment scheme.

350.2 414.0555.2

655.4

702.0

749.8

835.4

876.9

958.4

1042.0

1133.0

1184.0

1426.6

1464.6

1572.5

1772.7

1873.7

1918.0

2017.6

2059.8

JC_SW1116_NG_2_020312.D: -MS2(1213.4), 38.6min #1479

0

1

2

3

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 m/z

+

Glycan 158 Parent ion: m/z 1213.62- Composition: (Hex)2 (HexNAc)3 (Deoxyhexose)2 (NeuAc)1 + (Man)3(GlcNAc)2

Notes: Distinction between 3-arm/6-arm not intended in glycan fragment scheme. Additional isomers of fragments cannot be excluded.

350.3

425.2

467.0

570.2

655.2

835.4

884.9

965.9

1039.5

1200.5

1388.1 1474.51566.6

1751.7

1889.7

1992.8

2074.6

2153.8

-MS2(1221.5), 48.2min #1849

0.0

0.5

1.0

1.5

4x10

Intens.

400 600 800 1000 1200 1400 1600 1800 2000 2200 m/z

Notes: Distinction between 3-arm/6-arm not intended in glycan fragment sch