Standardization of an Immunoassay for the Detection of Antibodies to B2 Glycoprotein-I

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Erickson, Eric N., Jr.
Immunoassay; Antibodies; B2 Glycoprotein-I; Biology; Diseases; Immune System Diseases; Life Sciences
thesis / dissertation description
Antiphospholipid Syndrome (APS) is an evolving autoimmune disease with numerous clinical manifestations. APS occurs in two forms: Primary Antiphospholipid Syndrome (PAPS) and secondarily in association with other autoimmune disorders such as Systemic Lupus Erythematosis (SLE). In the brief period since its discovery as a cofactor for anticardiolipin antibodies, p1-glycoprotein I (�GPI; apolipoprotein H) has been recognized as the autoantigen in the absence of anionic phospholipid when appropriately presented to human autoantibodies. Recent studies suggest that this presentation requires the surface-dependent unmasking of a cryptic epitope. Immunoassays of autoantibodies directed against p1GPI, developed through the application of suitable polymeric matrices, may be clinicaJly relevant in that they may be more specific for (APS). Assays of Anti-� Glycoptotein I (aj32GPI) have used a variety of materials in the antigen-specific presentation system. This situation renders this assay particularly susceptible to interlaboratory variation when raw absorbance data are reported. Presently, many assay systems for the detection of APS lack standardization and are plagued with technical problems. Reference materials for a uniform enzyme immunoassay oflgG aj31GPI have not, as yet, been available. The following thesis focuses on the development and standardization of a clinically significant assay for the detection of APS. The assay centers on the detection of a�GPI antibodies. Described herein, is a method utilizing commercially available, chemically activated polystyrene microtiter plates with a surface -which binds proteins covalently. Additionally, a commercial source of human P,GPI has been obtained. been Reference calibrators have developed consisting of a control group of 111 men and 93 women, yielding a range of 0-19 standard lgG antj.j.\GPI units (SGU) for the ninety-fifth percentile of values. These are intended as interim calibrators that can be used to develop primary international standards for an immunoassay of lgO aP2GPI.