title: Mass spectrometry analyses of κ and λ fractions result in increased number of complementarity-determining region identifications.
abstract: Sera from lung cancer patients contain antibodies against tumor-associated antigens. Specific amino acid sequences of the complementarity-determining regions (CDRs) in the antigen-binding fragment (Fab) of these antibodies have potential as lung cancer biomarkers. Detection and identification of CDRs by mass spectrometry can significantly be improved by reduction of the complexity of the immunoglobulin molecule. Our aim was to molecular dissect IgG into κ and λ fragments to reduce the complexity and thereby identify substantially more CDRs than by just total Fab isolation. We purified Fab, Fab-κ, Fab-λ, κ and λ light chains from serum from 10 stage I lung adenocarcinoma patients and 10 matched controls from the current and former smokers. After purification, the immunoglobulin fragments were enzymatically digested and measured by high-resolution mass spectrometry. Finally, we compared the number of CDRs identified in these immunoglobulin fragments with that in the Fab fragments. Twice as many CDRs were identified when Fab-κ, Fab-λ, κ and λ (3330) were combined than in the Fab fraction (1663) alone. The number of CDRs and κ:λ ratio was statistically similar in both cases and controls. Molecular dissection of IgG identifies significantly more CDRs, which increases the likelihood of finding lung cancer-related CDR sequences.
authors: Department of Clinical Chemistry, Erasmus Medical Center, Rotterdam, The Netherlands.
authors: Ingrid
authors: I
authors: Broodman
authors: Dominique
authors: D
authors: de Costa
authors: Christoph
authors: C
authors: Stingl
authors: Lennard J M
authors: LJ
authors: Dekker
authors: Martijn M
authors: MM
authors: VanDuijn
authors: Jan
authors: J
authors: Lindemans
authors: Rob J
authors: RJ
authors: van Klaveren
authors: Theo M
authors: TM
authors: Luider
mesh_terms: D000230:Adenocarcinoma / Q000097:blood; D000077192:Adenocarcinoma of Lung; D000368:Aged; D016022:Case-Control Studies; D022801:Complementarity Determining Regions / Q000032:analysis / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007145:Immunoglobulin kappa-Chains / Q000032:analysis* / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification; D007146:Immunoglobulin lambda-Chains / Q000032:analysis* / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification; D008175:Lung Neoplasms / Q000097:blood; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D008875:Middle Aged; D016017:Odds Ratio; D016032:Randomized Controlled Trials as Topic; D015203:Reproducibility of Results; D012907:Smoking / Q000097:blood
chemical_list: D022801:Complementarity Determining Regions; D007145:Immunoglobulin kappa-Chains; D007146:Immunoglobulin lambda-Chains