Supplemental Data
Table S1: Patient Information.
Fig A: Apolipoprotein C1 ImmunoSELDI. The experiment was performed as descibed in reference 7. The antibody used was a mouse monoclonal to human apolipoprotein C1 purchased from Chemicon International Inc (MAB1064).
Fig B: Lsozyme C ImmunoSELDI. The antibody used was a rabbit polyclonal to human lysozyme C purchased from Abcam (ab2408)
Figs C & D: MS/MS spectra used to identify Lysozyme
Fig E-I: MS/MS spectra used to identify Transthyretin
Fig J: Transthyretin ImmunoSELDI. The antibody used was a rabbit polyclonal to human transthyretin purchased from from Abcam (ab16006).
Figs K & L: MS/MS spectra used to identify th4 m/z 5080 fragment of transthyretin.
Fig M: Accurate mass determination of the m/z 5080 fragment of transthyretin.
Fig N: M/Z 5080 ImmunoSELDI. The same antibody was used as in the intact transthyretin ImmunoSELDI (Fig J).
Fig O: MS/MS spectra used to identify the m/z 3273 and 3289 fragments of ITIH4. The top spectrum/peptide is common to both fragments and the annotated spectra below show both the oxidised and non-oxidised verions of the methioinine containing peptide.
Rathinam et al: Changes in the serum proteome associated with lung cancer and their reversal following tumor resection
6200 6400 6600 6800
6200 6400 6600 6800
IMAC follow up-A
IMAC follow up-B
IMAC follow up-C
*
Wholeserum
Depletedserum
Elutedproteins
Apolipoprotein C1mmunoSELDI
Inte
nsity
m/z
*
Fig A
Fig B 13000 14000 15000 16000
13000 14000 15000 16000
imaclowrange-D
imaclowrange-E
imaclowrange-F
13000 14000 15000 16000
13000 14000 15000 16000
m/z
Inte
nsity
A
B
C
*
*
Lysozyme C ImmunoSELDI
Wholeserum
Depletedserum
Elutedproteins
Lysozyme MS/MS
R.STDYGIFQINSR.Y
R.GISLALWMC*LAK.W* = carboxyamidomethylation
Fig C
Fig D
Transthyretin MS/MS
R.GSPAINVAVHVFR.KFig F
K.TSESGELHGLTTEEEFVEGIYK.VFig E
K.AADDTWEPFASGK.TFig G
R.KAADDTWEPFASGK.TFig H
K.ALGISPFHEHAEVVFTANDSGPR.RFig I
13000 13500 14000 14500 15000
13000 13500 14000 14500 15000
no antibody f t
plus antibody f t
plus antibody eluent
13000 14000 15000
m/z
Inte
nsity
A
B
C
Fig JTransthyretin ImmunoSELDI
Whole serum
Depleted serum
Elutedproteins
Fig K K.ALGISPFHEHAEVVFTANDSGPR.R
‘5080’ MS/MS
A.ALLSPYSYSTTAVVTNPK.EFig L
M2
M1
5050 5075 5100 5125
5050 5075 5100 5125
dil
dep
imaclowrange-F
*
Wholeserum
Depletedserum
Elutedproteins
m/z 5080(Transthyretin Fragment)ImmunoSELDI
Inte
nsity
m/z
Fig M
Fig N
‘3273/3289’ MS/MS
Fig OR.PGVLSSRQLGLPGPPDVPDHAAYHPF.R
(common to both)
MNFRPGVLSSR
M*NFRPGVLSSR
y1-2
y1-3
y1-4
b1-2
b1-4
y1-6 y1
-7 b1-7
y1-3
y1-4
b1-2
b1-4
y1-6
y1-7
b1-7