A chimeric antibody with the human γ1 constant region as a putative standard for assays to detect IgG β2-glycoprotein I-dependent anticardiolipin and anti-β2-glycoprotein I antibodies

Kenji Ichikawa, Akito Tsutsumi, Tatsuya Atsumi, Eiji Matsuura, Seiichi Kobayashi, Graham R.V. Hughes, Munther A. Khamashta, Takao Koike

Research output: Contribution to journalArticle

89 Citations (Scopus)

Abstract

Objective. Thromboembolic manifestations or thrombocytopenia in association with anticardiolipin antibodies (aCL) or lupus anticoagulant are known as the antiphospholipid syndrome (APS). Efforts have been made to elucidate precise clinical features and adequate therapeutic options for treating patients with APS. However, the lack of a proper international standard for measurement of aCL makes it difficult to compare data derived from different laboratories. We attempted to design a chimeric antibody with human γ constant regions and variable regions of WBCAL-1, a monoclonal antibody established from an APS-prone mouse which has a specificity similar to that of aCL in sera from humans with APS. Methods. Variable-region genes of WBCAL-1, which were cloned using reverse transcription-polymerase chain reaction, were inserted into plasmids containing human γ1 and κ constant- region genes. The construct was transfected to a mouse myeloma cell line. Stable transfectants that secreted a chimeric antibody, HCAL, into the culture supernatant were obtained. The reactivity of HCAL to cardiolipin and to β2-glycoprotein I (β2GPI) was studied using a solid-phase enzyme immunoassay. The binding of HCAL was compared with the binding of standards for IgG aCL and anti-β2GPI antibody assays done in 18 independent laboratories. Results. In the presence of β2GPI, HCAL bound to the wells of cardiolipin-coated microtiter plates in a dose-dependent manner and reacted with β2GPI on oxygenated polystyrene plates. The aCL activity of HCAL can be converted into GPL units (IgG phospholipid units), which is widely used to quantify IgG aCL activity, using the following formula: 1 GPL unit = 32.9 x (concentration of HCAL [in μg/ml])0.503. The reactivity of HCAL to cardiolipin or β2GPI was similar to the reactivity of standards for IgG aCL or anti-β2GPI antibody assays done in collaborative laboratories. Conclusion. Because the reactivity of HCAL is similar to that of aCL in sera from humans with APS, HCAL will be useful as a standard for human IgG aCL and anti-β2GPI antibody assays.

Original languageEnglish
Pages (from-to)2461-2470
Number of pages10
JournalArthritis and Rheumatism
Volume42
Issue number11
DOIs
Publication statusPublished - Nov 1999

ASJC Scopus subject areas

  • Immunology and Allergy
  • Rheumatology
  • Immunology
  • Pharmacology (medical)

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