IRBP: preparation and characterization of site-specific monoclonal antibodies
作者:
DonosoLarry A.,
RodriguesMerlyn,
VrabecTamara R.,
SeryTheodore W.,
LingShao,
期刊:
Current Eye Research
(Taylor Available online 1990)
卷期:
Volume 9,
issue 4
页码: 357-362
ISSN:0271-3683
年代: 1990
DOI:10.3109/02713689008999623
出版商: Taylor&Francis
数据来源: Taylor
摘要:
Interstitial retinoid binding protein (IRBP) is a 136,000 molecular weight photoreceptor cell protein which is a highly pathogenic autoantigen for the induction of experimental autoimmune uveitis (EAU). In this study we produced a series of monoclonal antibodies (MAbs) which define different epitopes in the native molecule. These MAbs were further subdivided into three distinct groups based on a radioimmunoassay, and by ELISA assay using native IRBP and synthetic peptides corresponding to its entire amino acid sequence. Group I MAbs (MAbD7-B1 and MAbC6-B4) bound to native IRBP but not to any synthetic peptides, suggesting that their antigenic epitopes are strictly conformation dependent. Group II MAbs (MAbC7-D3 and MAbG8-H4) bound weakly to multiple peptides which shared amino acid sequence similarity located within each of four homology domains indicating that these epitopes are also conformation dependent. In group III (MAbH3-B5, MAbH7-A5, and MAbB6-D12) MAb binding was localized to a specific peptide. The MAbH3-B5 binding site was further refined to amino acid positions 361 to 367 in the native molecule. MAbH3-B5 was also useful in localizing IRBP in the mouse retina by immuno-histochemical techniques. The application of these MAbs in the study of EAU and interphotoreceptor transport mechanisms is discussed.
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