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免疫过氧化物酶技术在针对血管性血友病因子(vWf)的单克隆抗体(Mabs)特异性检测中的应用。

Applications of immunoperoxidase techniques in specificity testing of monoclonal antibodies (Mabs) against von Willebrand factor (vWf).

作者信息

Ingerslev J, Bukh A, Møller N P, Stenbjerg S, Zeuthen J

机构信息

Department of Clinical Immunology, University Hospital Aarhus, Denmark.

出版信息

Clin Chim Acta. 1988 May 13;174(1):65-81. doi: 10.1016/0009-8981(88)90367-1.

Abstract

The present communication describes the production of a new series of murine Mabs against von Willebrand factor (vWf) in which specificity was tested using immunoperoxidase techniques. Seven Mabs showed specific reactivity with native and disaggregated vWf, whereas no binding was found to material from patients with severe homozygous (or doubly heterozygous) von Willebrand's disease (vWd) or factor VIII coagulant antigen (VIII:Ag). These Mabs are thought to carry separate specificities as only slight or no competitive activity was detected. Four Mabs partially inhibited the ristocetin-induced platelet agglutination and three interacted with vWf-binding to type I collagen. All antibodies bound to the complete range of vWf multimers of normal plasma. Excellent binding and detection properties of Mabs were found in asymmetrical two-site enzyme linked immunosorbent assays (ELISA) for quantitation of vWf antigen (vWf:Ag). One particular antibody (Mab vWf-33) discriminated vWf material from a number of subtype II vWd plasmas tested.

摘要

本报告描述了一系列针对血管性血友病因子(vWf)的新型鼠单克隆抗体(Mabs)的制备过程,其中利用免疫过氧化物酶技术检测了其特异性。七种单克隆抗体与天然和去聚集的vWf表现出特异性反应,而未发现与重度纯合子(或双重杂合子)血管性血友病(vWd)患者的物质或凝血因子VIII凝血抗原(VIII:Ag)有结合。由于仅检测到轻微或无竞争活性,这些单克隆抗体被认为具有不同的特异性。四种单克隆抗体部分抑制了瑞斯托霉素诱导的血小板凝集,三种单克隆抗体与vWf与I型胶原的结合相互作用。所有抗体均与正常血浆中完整范围的vWf多聚体结合。在用于定量vWf抗原(vWf:Ag)的不对称双位点酶联免疫吸附测定(ELISA)中,发现单克隆抗体具有出色的结合和检测特性。一种特定抗体(单克隆抗体vWf - 33)区分了所测试的多种II型vWd血浆中的vWf物质。

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