Mass spectrometry analyses of ? and ? fractions result in increased number of complementarity-determining region identifications
| Autor: | Martijn M. VanDuijn, Theo M. Luider, Dominique de Costa, Jan Lindemans, Lennard J. M. Dekker, Ingrid Broodman, Rob J. van Klaveren, Christoph Stingl |
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| Přispěvatelé: | Clinical Chemistry, Pulmonary Medicine, Neurology |
| Rok vydání: | 2012 |
| Předmět: |
Male
Lung Neoplasms Adenocarcinoma of Lung Complementarity determining region Adenocarcinoma Mass spectrometry Immunoglobulin light chain Biochemistry Mass Spectrometry Immunoglobulin kappa-Chains Immunoglobulin lambda-Chains Antigen SDG 3 - Good Health and Well-being Fab Fragments Odds Ratio Humans Molecular Biology Aged Randomized Controlled Trials as Topic chemistry.chemical_classification biology Chemistry Smoking Reproducibility of Results Middle Aged Complementarity Determining Regions Molecular biology Amino acid Case-Control Studies Immunoglobulin Fragments biology.protein Electrophoresis Polyacrylamide Gel Female Antibody |
| Zdroj: | Proteomics, 12(2), 183-191. Wiley-VCH |
| ISSN: | 1615-9853 |
| DOI: | 10.1002/pmic.201100244 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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