Detection of antiphospholipid antibodies by immunostaining on thin layer chromatography plates
| Autor: | Sorice, Maurizio, Griggi, T., Circella, A., Garofalo, Tina, Dagostino, F., D'Agostino, F., Pittoni, V., Pontieri, Giuseppe, Lenti, Luisa, Valesini, Guido |
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| Rok vydání: | 1994 |
| Předmět: |
medicine.drug_class
Immunology Phospholipid Enzyme-Linked Immunosorbent Assay HIV Infections Monoclonal antibody Sensitivity and Specificity chemistry.chemical_compound Antigen medicine Humans Lupus Erythematosus Systemic Immunology and Allergy Syphilis Antigens Immunosorbent Techniques Phospholipids Immunoassay biology Chemistry Autoantibody Antibodies Monoclonal Tlc immunostaining Antiphospholipid Syndrome Molecular biology Thin-layer chromatography Evaluation Studies as Topic Antibodies Anticardiolipin Antibodies Antiphospholipid biology.protein immunostaining thin layer chromatography β2-glycoprotein i anti-cardiolipin antibody beta(2)-glycoprotein i antiphospholipid antibody anti-phospholipid antibody anticardiolipin antibody Chromatography Thin Layer Antibody Immunostaining |
| Zdroj: | Journal of Immunological Methods. 173:49-54 |
| ISSN: | 0022-1759 |
| DOI: | 10.1016/0022-1759(94)90282-8 |
| Popis: | There is increasing interest in the role of antiphospholipid antibodies in the so-called ‘antiphospholipid antibody syndrome’ (APS). The two major methods currently employed for detecting the autoantibodies are the solid phase ELISA and the LAI test (inhibition of phospholipid dependent coagulation assay). In our study we have tested the possibility of detecting antiphospholipid antibodies by immunostaining on thin layer chromatography (TLC) plates, since this technique permits the use of pure phospholipid molecules as antigen. Sera were collected from 20 patients with SLE without APS, 20 patients with APS, 20 anti-HIV positive subjects, ten patients with signs of APS but antiphospholipid negative (ELISA), 20 patients with syphilis and 40 matched blood donors. Results showed that only 72.3% of sera containing detectable levels of aCL antibodies in solid phase ELISA were also positive for aCL in TLC immunostaining; these discrepancies may be due to the presence of antibodies reacting with a protein complexed with phospholipid (β2-glycoprotein-I) or, alternatively, to the different antigenic presentation of phospholipids on chromatograms compared to the surface of microtitre wells. Furthermore, aCL monoclonal antibody CAL-3, as well as nine sera positive for aCL, also reacted with PS and PE. Previous absorption of these sera with CL micelles completely abolished the reactivity with PS and PE, demonstrating cross-reactivity among these three phospholipids. In conclusion, our findings reveal that TLC immunostaining is more specific, but less sensitive, than ELISA for the detection of antiphospholipid antibodies in human sera. |
| Databáze: | OpenAIRE |
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