Targeted protein quantitation and profiling using PVDF affinity probe and MALDI-TOF MS
Resource
PROTEOMICS 7 (17): 3038-3050
Journal
PROTEOMICS
Pages
3038-3050
Date Issued
2007
Date
2007
Author(s)
Chen, Shu-Hua
Liao, Hsin-Kai
Chang, Chih-Yang
Juo, Chiun-Gung
Chen, Jenn-Han
Chan, Sunney I.
Chen, Yu-Ju
Abstract
Development of a rapid, effective, and highly specific platform for target identification in complex biofluids is one of the most important tasks in proteomic research. Taking advantage of the natural hydrophobic interaction of PVDF with probe protein, a simple and effective method was developed for protein quantitation and profiling. Using antibody-antigen interactions as a proof-of-concept system, the targeted plasma proteins, serum amyloid P (SAP), serum amyloid A (SAA), and C-reactive protein (CRP), could be selectively isolated and enriched from human plasma by antibody-immobilized PVDF membrane and directly identified by MALDI-TOF MS without additional elution step. The approach was successfully applied to human plasma for rapid quantitation and variant screening of SAP, SAA, and CRP in healthy individuals and patients with gastric cancer. The triplexed on-probe quantitative analysis revealed significant overexpression of CRP and SAA in gastric cancer group, consistent with parallel ELISA measurements and pathological progression and prognostic significance reported in previous literatures. Furthermore, the variant mass profiling of the post-translationally modified forms revealed a high occurrence of de-sialic acid SAP in patients with gastric cancer. Due to the versatile assay design, ease of probe preparation without chemical synthesis, and compatibility with MALDI-TOF MS analysis, the methodology may be useful for target protein characterization, functional proteomics, and screening in clinical proteomics. ? 2007 Wiley-VCH Verlag GmbH & Co. KGaA.
Subjects
C-reactive protein; Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry; Quantitation; Serum amyloid A; Serum amyloid P
SDGs
Other Subjects
C reactive protein; serum amyloid A; serum amyloid P; sialic acid; antigen antibody reaction; article; controlled study; elution; enzyme linked immunosorbent assay; functional proteomics; human; hydrophobicity; matrix assisted laser desorption ionization time of flight mass spectrometry; methodology; normal human; priority journal; prognosis; protein isolation; protein targeting; quantitative analysis; screening; sensitivity and specificity; stomach cancer; synthesis; C-Reactive Protein; Case-Control Studies; Enzyme-Linked Immunosorbent Assay; Humans; Membranes, Artificial; Models, Biological; Polyvinyls; Prognosis; Protein Processing, Post-Translational; Proteomics; Serum Amyloid A Protein; Serum Amyloid P-Component; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Stomach Neoplasms
Type
journal article
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