Document 0538
 DOCN  M9480538
 TI    Detection of antiphospholipid antibodies by immunostaining on thin layer
       chromatography plates.
 DT    9410
 AU    Sorice M; Griggi T; Circella A; Garofalo T; d'Agostino F; Pittoni V;
       Pontieri GM; Lenti L; Valesini G; Dipartimento di Medicina Sperimentale,
       Universita La Sapienza,; Roma, Italy.
 SO    J Immunol Methods. 1994 Jul 12;173(1):49-54. Unique Identifier :
       AIDSLINE MED/94308563
 AB    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 (beta
       2-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.
 DE    Antibodies, Anticardiolipin/BLOOD  Antibodies, Antiphospholipid/*BLOOD
       Antibodies, Monoclonal  Antigens  Antiphospholipid Syndrome/IMMUNOLOGY
       Chromatography, Thin Layer/*METHODS/STATISTICS & NUMER DATA  Comparative
       Study  Enzyme-Linked Immunosorbent Assay/STATISTICS & NUMER DATA
       Evaluation Studies  Human  HIV Infections/IMMUNOLOGY
       Immunoassay/*METHODS/STATISTICS & NUMER DATA  Immunosorbent Techniques
       Lupus Erythematosus, Systemic/IMMUNOLOGY  Phospholipids/IMMUNOLOGY
       Sensitivity and Specificity  Support, Non-U.S. Gov't
       Syphilis/IMMUNOLOGY  JOURNAL ARTICLE

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