Monoclonal antibodies against different domains of human IgA: Specificities determined by immunoblotting and haemagglutination-inhibition
| Autor: | Arjen Vlug, Fred van Leeuwen, Gerda de Lange, Anita Faber, Peter van Eede, Roy Jefferis, Joost J. Haaijman, Jeike Biewenga |
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| Rok vydání: | 1986 |
| Předmět: |
biology
Myeloma protein medicine.drug_class Chemistry Immunology Antibodies Monoclonal Hemagglutination Inhibition Tests Immunoglobulin light chain Monoclonal antibody Primary and secondary antibodies Molecular biology Epitope Immunoglobulin A Haemagglutination inhibition Epitopes Antibody Specificity Monoclonal biology.protein medicine Humans Electrophoresis Polyacrylamide Gel Antibody Immunoglobulin Fragments Molecular Biology |
| Zdroj: | Molecular Immunology. 23:761-767 |
| ISSN: | 0161-5890 |
| DOI: | 10.1016/0161-5890(86)90088-x |
| Popis: | The specificity of 14 monoclonal antibodies has been determined by immunoblotting (IB) and haemagglutination-inhibition (HAI) analysis using IgA1 and IgA2 myeloma proteins and eight different IgA1 fragments. Two antibodies probably recognized epitopes on the CH1 domain of IgA. They reacted with all Fab-containing fragments irrespective of whether these originated from the same or different IgA proteins. Seven antibodies were directed against epitopes on the CH2 domain. These antibodies were reactive with F(abc)2 fragments. They failed to react with Fab, Fab' and F(ab')2 fragments. Two out of these seven antibodies did not react with two-chain IgA half-molecules and Fabc fragments containing a single heavy and a single light chain. This suggests that these two antibodies recognized an epitope whose structure is dependent on disulfide linked heavy chains. Five other antibodies showed specificity for the CH3 domain. They were reactive with all CH3-containing molecules, irrespective of whether they comprised one or two a chains. Our study demonstrates that IB is an appropriate technique to determine domain specificity of monoclonal anti-immunoglobulin reagents. Although the IB tests were performed on denatured proteins the results agreed surprisingly well with those of the HAI analyses. Moreover, the IB technique could be used on fragments which could not be purified well enough for HAI analyses. |
| Databáze: | OpenAIRE |
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