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Binding of high-avidity anti-[beta]2-glycoprotein I antibodiesČučnik, Saša ...Objectives. We studied the influence of different binding conditions on the interaction between high- or low-avidity IgG anti-beta2-glycoprotein I antibodies (anti-beta2-GPI) and beta2-GPI, which was ... either free in solution or bound to microtitre plates or nitrocellulose membranes. Methods. Sera from 30 patients with antiphospholipid syndrome and/or systemic lupus erythematosuswere selected on the basis of anti-beta2-GPI positivity. Avidity of IgG anti-beta2-GPI was determined by chaotropic ELISA, using increased NaCIconcentration during the antibody binding. Immunodetection on nitrocellulose membrane followed reducing or non-reducing sodium dodecyl sulphate-polyacrylamide gel electrophoresis (PAGE) of beta2-GPI. In converted,non-reducing PAGE, the preparation with high-affinity Fab fragments (obtained by papain digestion) was subjected to electrophoresis, while purified beta2-GPI was used as the sample in immunodetection. Results. Anti-beta2-GPI antibodies of high, low or heterogeneous (low and high) aviditywere found in 5/30, 9/30 and 16/30 sera, respectively. The density of beta2-GPI, which was 20-30 times higher on the nitrocellulose membrane than onthe surface of ELISA plates, was not sufficient for the recognition of the antigen by anti-beta2-GPI: 2/5 high-avidity samples reacted only with non-reduced beta2-GPI, 3/9 low-avidity samples recognized only denatured and reduced beta2-GPI, and 1/16 samples with heterogeneous-avidity antibodies reacted with reduced and non-reduced beta2-GPI. Conclusions. Our results suggest that neither high density of the antigen nor high avidity of the antibodies (or Fab fragments) alone was sufficient for the binding of anti-beta2-GPI to beta2-GPI. Some conformational modifications and, consequently, exposed neo-epitopes are required for the recognition of beta2-GPI by polyclonal anti-beta2-GPI antibodies.Vir: Rheumatology. - ISSN 1462-0324 (Letn. 43, 2004, str. 1353-1356)Vrsta gradiva - članek, sestavni delLeto - 2004Jezik - angleškiCOBISS.SI-ID - 18763993
Avtor
Čučnik, Saša |
Kveder, Tanja |
Rozman, Blaž, 1944- |
Božič, Borut, 1959-
Teme
Antiphospholipid Syndrome |
Lupus Erythematosus, Systemic |
Autoantibodies |
IgG |
Glycoproteins |
Electrophoresis, Polyacrylamide Gel |
Enzyme-Linked Immunosorbent Assay |
Antibody Affinity |
Immunoglobulins, Fab |
Antifosfolipidni sindrom |
Avtoprotitelesa |
Glikoproteini |
IgG |
Lupus eritematozni, sistemski |
Elektroforeza, poliakrilamid gel |
Encimsko-vezani imunosorbentni test |
Imunoglobulini, Fab |
Protitelo, afiniteta
Vnos na polico
Trajna povezava
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Leto | Faktor vpliva | Izdaja | Kategorija | Razvrstitev | ||||
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JCR | SNIP | JCR | SNIP | JCR | SNIP | JCR | SNIP |
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Povezave do osebnih bibliografij avtorjev | Povezave do podatkov o raziskovalcih v sistemu SICRIS |
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Čučnik, Saša | 16362 |
Kveder, Tanja | 02066 |
Rozman, Blaž, 1944- | 07180 |
Božič, Borut, 1959- | 00781 |
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