CE-based noncompetitive immunoassay for immunoglobulin G in bovine colostrum products

A CE-based noncompetitive immunoassay for IgG in bovine colostrum products was established. FITC-labeled protein G (FITC-PrG) was tagged through noncovalent bindings to the Fc region of the mouse monoclonal antibovine IgG (Ab). The FITC-PrG, Ab, and IgG formed a sandwiched immunocomplex FITC-PrG-Ab-...

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Bibliographic Details
Published in:Electrophoresis 2007-11, Vol.28 (21), p.3934-3939
Main Authors: Zhao, Jin, Ding, Xiaojing, Wang, Xinyu, Wang, Qian, Wang, Zhi
Format: Article
Language:English
Subjects:
Animals
Antibody Affinity
Bacterial Proteins - chemistry
Binding Sites, Antibody
Biosensing Techniques
Bovine colostrum
Cattle
Colostrum - chemistry
Electrophoresis, Capillary - methods
Fluorescein-5-isothiocyanate - chemistry
Immunoassay - methods
Immunoglobulin G
Immunoglobulin G - analysis
Lasers
LIF detection
Mice
Noncompetitive immunoassay
Sensitivity and Specificity
Spectrometry, Fluorescence
Spectrophotometry, Ultraviolet
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Summary:A CE-based noncompetitive immunoassay for IgG in bovine colostrum products was established. FITC-labeled protein G (FITC-PrG) was tagged through noncovalent bindings to the Fc region of the mouse monoclonal antibovine IgG (Ab). The FITC-PrG, Ab, and IgG formed a sandwiched immunocomplex FITC-PrG-Ab-IgG under optimal incubation conditions. The immunocomplex was separated and analyzed by CZE with LIF detection in less than 2 min in an uncoated fused-silica capillary. Addition of PEG 20 000 (PEG 20M) in the running buffer significantly suppressed analyte adsorption and thus improved the reproducibility and the resolution. The precision of the method was 5.1% (n = 7). A linear relationship was established for the IgG concentration in the range of 1-5 mg/L with a linear correlation coefficient (r = 0.9917). The LOD was 0.1 mg/L (S/N = 3). The method was successfully applied for the determination of IgG in bovine colostrum products and satisfactory results were achieved.
ISSN:0173-0835
1522-2683
DOI:10.1002/elps.200700122