IRBP: preparation and characterization of site-specific monoclonal antibodies

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 epitope...

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Bibliographic Details
Published inCurrent eye research Vol. 9; no. 4; p. 357
Main Authors Donoso, L A, Rodrigues, M, Vrabec, T R, Sery, T W, Fong, S L
Format Journal Article
LanguageEnglish
Published England 1990
Subjects
Amino Acid Sequence
Animals
Antibodies, Monoclonal - biosynthesis
Antibodies, Monoclonal - immunology
Antibody Specificity
Binding Sites, Antibody - immunology
Binding, Competitive
Epitopes - immunology
Eye Proteins
Humans
Immunoenzyme Techniques
Mice
Mice, Inbred BALB C
Molecular Sequence Data
Peptides - chemical synthesis
Peptides - immunology
Retinol-Binding Proteins - immunology
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Summary: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 immunohistochemical techniques. The application of these MAbs in the study of EAU and interphotoreceptor transport mechanisms is discussed.
ISSN:0271-3683
DOI:10.3109/02713689008999623