Peptides from the variable region of specific antibodies are shared among lung cancer patients.
Late diagnosis of lung cancer is still the main reason for high mortality rates in lung cancer. Lung cancer is a heterogeneous disease which induces an immune response to different tumor antigens. Several methods for searching autoantibodies have been described that are based on known purified antigen panels. The aim of our study is to find evidence that parts of the antigen-binding-domain of antibodies are shared among lung cancer patients. This was investigated by a novel approach based on sequencing antigen-binding-fragments (Fab) of immunoglobulins using proteomic techniques without the need of previously known antigen panels. From serum of 93 participants of the NELSON trial IgG was isolated and subsequently digested into Fab and Fc. Fab was purified from the digested mixture by SDS-PAGE. The Fab containing gel-bands were excised, tryptic digested and measured on a nano-LC-Orbitrap-Mass-spectrometry system. Multivariate analysis of the mass spectrometry data by linear canonical discriminant analysis combined with stepwise logistic regression resulted in a 12-antibody-peptide model which was able to distinguish lung cancer patients from controls in a high risk population with a sensitivity of 84% and specificity of 90%. With our Fab-purification combined Orbitrap-mass-spectrometry approach, we found peptides from the variable-parts of antibodies which are shared among lung cancer patients.
Department of Pulmonology, Erasmus Medical Center, Rotterdam, The Netherlands.
Dominique
D
de Costa
Department of Clinical Chemistry, Erasmus Medical Center, Rotterdam, The Netherlands.
Ingrid
I
Broodman
StatistiCal BV, Wassenaar, The Netherlands.
Wim
W
Calame
Department of Neurology, Erasmus Medical Center, Rotterdam, The Netherlands.
Christoph
C
Stingl
Department of Neurology, Erasmus Medical Center, Rotterdam, The Netherlands.
Lennard J M
LJ
Dekker
Unit of Trials and Statistics, Erasmus Medical Center, Rotterdam, The Netherlands.
René M
RM
Vernhout
Unit of Public Health, Erasmus Medical Center, Rotterdam, The Netherlands.
Harry J
HJ
de Koning
Department of Pulmonology, Erasmus Medical Center, Rotterdam, The Netherlands.
Henk C
HC
Hoogsteden
Department of Neurology, Erasmus Medical Center, Rotterdam, The Netherlands.
Peter A E
PA
Sillevis Smitt
Department of Pulmonology, Erasmus Medical Center, Rotterdam, The Netherlands.
Rob J
RJ
van Klaveren
Department of Neurology, Erasmus Medical Center, Rotterdam, The Netherlands.
Theo M
TM
Luider
Department of Neurology, Erasmus Medical Center, Rotterdam, The Netherlands.
Martijn M
MM
Vanduijn
D000368:Aged; D000595:Amino Acid Sequence; D000906:Antibodies / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000276:immunology*; D016022:Case-Control Studies; D002853:Chromatography, Liquid; D005260:Female; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000737:chemistry / Q000276:immunology*; D008175:Lung Neoplasms / Q000175:diagnosis / Q000276:immunology* / Q000473:pathology; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D008969:Molecular Sequence Data; D009367:Neoplasm Staging; D010455:Peptides / Q000737:chemistry / Q000276:immunology*; D015203:Reproducibility of Results
D000906:Antibodies; D000951:Antigens, Neoplasm; D007140:Immunoglobulin Fab Fragments; D007135:Immunoglobulin Variable Region; D010455:Peptides