Document Detail

Production of immunoaffinity membranes for direct analysis of antigen after antibody separation and blotting under non-denaturing conditions.
MedLine Citation:
PMID:  19446064     Owner:  NLM     Status:  MEDLINE    
Membrane-immobilized anti-transferrin antibody, which was produced after antibody was separated using non-denaturing two-dimensional electrophoresis (2DE), was transferred to a polyvinylidene difluoride (PVDF) membrane and was stained by direct blue 71. The antigen, transferrin, specifically bound to the membrane-immobilized antibody and was eluted only after rinsing the membrane with glutamic acid or aspartic acid solution. The antigen was analyzed directly by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) when the membrane was incubated with human plasma after the removal of human serum albumin using polyethylene glycol. The transferrin extracted by glutamic acid or aspartic acid solution retained the binding of Fe3+ in the presence of the carbonate anion. We found that immunoaffinity membranes can be useful for simple and rapid removal and extraction of intact proteins after antibody separation and blotting under non-denaturing conditions.
Youji Shimazaki; Azusa Kodama
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Publication Detail:
Type:  Journal Article     Date:  2009-04-11
Journal Detail:
Title:  Analytica chimica acta     Volume:  643     ISSN:  1873-4324     ISO Abbreviation:  Anal. Chim. Acta     Publication Date:  2009 Jun 
Date Detail:
Created Date:  2009-05-18     Completed Date:  2009-07-17     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370534     Medline TA:  Anal Chim Acta     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  61-6     Citation Subset:  IM    
Graduate School of Science and Engineering (Science Section) and Venture Business Laboratory, Ehime University, Bunkyo-cho 2-5, Matsuyama City 790-8577, Japan.
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MeSH Terms
Antibodies / immunology,  isolation & purification*
Antigen-Antibody Reactions / immunology
Antigens / analysis*,  immunology
Aspartic Acid / chemistry
Electrophoresis, Gel, Two-Dimensional
Glutamic Acid / chemistry
Immunosorbent Techniques*
Membranes, Artificial
Polyvinyls / chemistry*
Sensitivity and Specificity
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Transferrin / analysis,  immunology
Reg. No./Substance:
0/Antibodies; 0/Antigens; 0/Membranes, Artificial; 0/Polyvinyls; 11096-37-0/Transferrin; 24937-79-9/polyvinylidene fluoride; 56-84-8/Aspartic Acid; 56-86-0/Glutamic Acid

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