Methods to quantitate human haptoglobin by complexation with hemoglobin.
Native human haptoglobin isolated from normal human plasma by affinity chromatography on chicken hemoglobin -Sepharose was used as standard antigen. A direct sandwich ELISA for haptoglobin was developed, with human hemoglobin as a capturing agent. The peroxidase activity of the complex was measured as a means of detecting functional haptoglobin. The reactivities of monoclonal antibody vs. polyclonal antisera on the haptoglobin-hemoglobin complex were compared. Adopting a monoclonal antibody, clone 21.7, which is directed to the alpha chain of haptoglobin, a specific method to quantitate the native haptoglobin which can complex with hemoglobin has been developed.
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Department of Microbiology, Boston University School of Medicine, MA 02118.
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D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D001201:Ascites / Q000097:blood; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006242:Haptoglobins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D006258:Head and Neck Neoplasms / Q000097:blood; D006454:Hemoglobins / Q000737:chemistry*; D006801:Humans; D007124:Immunoenzyme Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D008875:Middle Aged; D010051:Ovarian Neoplasms / Q000097:blood
D000911:Antibodies, Monoclonal; D006242:Haptoglobins; D006454:Hemoglobins