Co-crystallization – tripartite:

results

Stacey Gulde

HWI

water400 20%

400 40%

400 80%

1000 20%

1000 40%

4000 20%

4000 40%

8000 20%

8000 40%

20,000 20%

20,000 40%

water

4567

7.589

PEGS

Bu

ffe

rs (

pH

)

Crystallization Cocktails10% (v/v)

Additive: Native-Buffer, DMSO or CoC

7

2 3 4

5 6

1

8

9 10 11 12

13 14 15 16

Native-Buffer DMSO

CoC #1

CoC #2

CoC #3

CoC #4

CoC #5

CoC #6

CoC #7

CoC #8

CoC #9

CoC #10

CoC #11

CoC #12

CoC #13

CoC #14

+90% (v/v)

PEG/pH-Buffer

Dispensed into a 1536 Plate w/ oil

Compounds

SGPP-0010001SGPP-0010003SGPP-0010004SGPP-0010006SGPP-0010007SGPP-0010010SGPP-0010011SGPP-0010013SGPP-0010015SGPP-0010016SGPP-0010017SGPP-0010020SGPP-0010026

Co-Crystallants Proteins ExaminedLysozyme

Laet002135AABLbra00619AAC

Lguy003107AAALinf002302AAALinf004718AAALinf007771AAALinf008585AAA

Lmaj002639AAALmaj002697AAA

Crystallization Drop Solubility (w/Protein)

50% (v/v) Protein & 50% (v/v) Crystallization cocktail3/10 proteins - independent of CoC

• Lysozyme: 7% 7%• Laet002135AAB: 85% 1%• Lbra00619AAC: 72% 1%

7/10 proteins – dependent upon CoC• Linf004718AAA: 36% - 98% ppt• Linf007771AAA: 12% – 100% ppt

Of the 14 Co-crystallants

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

Linf004718AAA Linf007771AAAProtein

% p

pt

Native-Buffer

DMSO

SGPP-0010001

SGPP-0010003

SGPP-0010004

SGPP-0010006

SGPP-0010007

SGPP-0010010

SGPP-0010011

SGPP-0010013

SGPP0010015

SGPP-0010016

SGPP-0010017

SGPP-0010019

SGPP-0010020

SGPP-0010026

Variation of Extent of Precipitation

If ≥ 90% ppt, then considered completely ppt

0

1

2

3

4

5

6

7

8

9

10

Additive

# P

rote

ins

Pp

t C

om

ple

tely

Extent of Protein Precipitation

Variation of Co-Crystallant Effect on

Protein Solubility

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

SGPP-0010001 SGPP-0010010Co-Crystallant

% p

pt

Lysozyme

Laet002135AAB

Lbra00619AAC

Lguy003107AAA

Linf002302AAA

Linf004718AAA

Linf007771AAA

Linf008585AAA

Lmaj002639AAA

Lmaj002697AAA SeMet