Anti-peptide reagent identifies a primary-structure-dependent, cross-reactive idiotype expressed on heavy and light chains from a murine monoclonal anti-CD4.
A synthetic peptide corresponding to the second complementarity determining region (CDR2) of the immunoglobulin (Ig) variable (V) region heavy (H) chain (CDR2VH) of anti-Leu3a, a murine monoclonal antibody specific for the human CD4 molecule, was used to elicit the production of specific rabbit anti-peptide antibodies. The rabbit anti-peptide antiserum was tested for reactivity against the immunizing peptide, anti-Leu3a, and a panel of mouse monoclonal anti-CD4. Only the immunizing peptide and anti-Leu3a were recognized by ELISA, whereas the H chains of anti-Leu3a and five other monoclonal anti-CD4 preparations were recognized by Western blot analysis. These data suggest that linear structures corresponding to the CDR2VH are not normally exposed on the surface of these monoclonal antibodies and become accessible only upon unfolding of the Ig molecule. In addition, Western blot analysis demonstrated that the anti-CDR2VH peptide antiserum was able to recognize the Ig light (L) chain of anti-Leu3a. This reactivity to both H and L chains from anti-Leu3a was ascribed to a homologous five amino acid sequence region shared by the two chains. The region of homology was associated with the third framework (FR3) of the L chain and was included as a portion of the sequence in the CDR2VH synthetic peptide. This observation was confirmed by the ability of the CDR2VH anti-peptide antiserum to bind the L chains of three mouse myeloma proteins that exhibited the five amino acid sequence region of homology within their respective FR3. Together, these data provide information on the structural basis of idiotypes shared by the H and L chains from the same antibody molecule and indicate that five amino acids might be sufficient to define a minimal continuous idiotypic determinant.
Attanasio
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R
Department of Virology and Immunology, Southwest Foundation for Biomedical Research, San Antonio, TX 78228-0147.
Kanda
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Stunz
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GW
Buck
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DW
Kennedy
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D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D015153:Blotting, Western; D015704:CD4 Antigens / Q000276:immunology*; D003429:Cross Reactions; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry*; D007130:Immunoglobulin Idiotypes / Q000276:immunology*; D007147:Immunoglobulin Light Chains / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D015394:Molecular Structure; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid
D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D007143:Immunoglobulin Heavy Chains; D007130:Immunoglobulin Idiotypes; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region