An ELISA method to measure total and specific human secretory IgA subclasses based on selective degradation by IgA1-protease
| Autor: | Michel D. Kazatchkine, Laurent Bélec, Jean-Pierre Bouvet, Hakim Hocini, S. Iscaki |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Adult
Secretory component medicine.medical_treatment Immunology Enzyme-Linked Immunosorbent Assay chemical and pharmacologic phenomena HIV Antibodies Immunoglobulin E Subclass HIV Envelope Protein gp160 Substrate Specificity fluids and secretions stomatognathic system Antibody Specificity HIV Seropositivity medicine Humans Immunology and Allergy Saliva chemistry.chemical_classification Protease biology Colostrum Serine Endopeptidases Isotype Molecular biology Enzyme chemistry Immunoglobulin A Secretory HIV-1 biology.protein Female Specific activity Antibody |
| Zdroj: | Journal of Immunological Methods. 235:53-60 |
| ISSN: | 0022-1759 |
| DOI: | 10.1016/s0022-1759(99)00214-8 |
| Popis: | We have taken advantage of the property of IgA1-proteases to selectively cleave the human IgA1 subclass into Fabalpha and Fcalpha-J chain-secretory component (Fcalpha-J-SC) fragments in order to design a novel ELISA method for measuring the two secretory IgA (S-IgA) subclasses in secretions. The assay is based on the loss of detection of S-IgA1 by a combination of peroxidase-labelled antibodies to secretory component and Fab following IgA1-protease treatment. The specificity is that of the protease and the sensitivity of the detection is 5 ng/ml. Moreover, the use of purified S-IgA1 and S-IgA2 controls is not necessary. The assay has been successfully applied to the analysis of colostral S-IgA antibodies (Abs) to HIV-1-gp160 from HIV-1 positive women. The major subclass of colostral S-IgA antibodies to gp160 was found to be of the alpha1 isotype but the specific activity of anti-HIV-gp160 S-IgA2 was, however, higher than that of S-IgA1. |
| Databáze: | OpenAIRE |
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