Hoegee-de Nobel E, Voskuilen M, Briët E, Brommer E J, Nieuwenhuizen W
Gaubius Institute TNO, Leiden, The Netherlands.
Thromb Haemost. 1988 Dec 22;60(3):415-8.
The most commonly used fibrinogen assays in the clinic are clotting rate assays, e.g. the Clauss method. Such functional assays may be disturbed by e.g. heparin, anticoagulant fibrinogen degradation products (FgDP) and in the case of a dysfibrinogenemia. Immunological methods would not suffer from these interferences. However, immunological assays for fibrinogen, which do not measure FgDPs, do not exist. To set up such an enzyme immunoassay (EIA) we developed two monoclonal antibodies. The first monoclonal antibody (G8) has its epitope in the carboxyl-terminal 150 amino acid stretches of the fibrinogen A alpha-chains. G8 is used to coat the wells of microtitration plates, and is the capture antibody in this EIA. The second antibody (Y18) has been described by us previously (Blood 1985; 66: 503). It is directed against fibrinopeptide A, covalently bound to the alpha-chains i.e. against the amino-terminal stretches of the A alpha-chains. Y18 is conjugated with horse-radish peroxidase, and used as tagging antibody. The EIA does not react with, and is not interfered by FgDP such as purified fragments X and Y, up to a concentration of 800 micrograms/ml. An FgDP mixture such as generated by Streptokinase treatment of plasma does not respond. Fibrin degradation products (whole blood lysate) up to 800 micrograms/ml do not interfere nor do heparin, EDTA or oxalate. The time-to-result of the EIA is only 45 minutes. Some patient plasmas yielded dose-response curves which are not parallel with the calibration curve of the EIA. An explanation for this phenomenon could not be given.(ABSTRACT TRUNCATED AT 250 WORDS)
临床上最常用的纤维蛋白原检测方法是凝血速率检测，例如Clauss法。此类功能检测可能会受到肝素、抗凝性纤维蛋白原降解产物（FgDP）等因素的干扰，在异常纤维蛋白原血症的情况下也是如此。免疫方法则不会受到这些干扰。然而，目前尚无用于检测纤维蛋白原且不检测FgDP的免疫检测方法。为建立这样一种酶免疫测定（EIA）方法，我们研发了两种单克隆抗体。第一种单克隆抗体（G8）的表位位于纤维蛋白原Aα链的羧基末端150个氨基酸片段。G8用于包被微量滴定板的孔，是该EIA中的捕获抗体。第二种抗体（Y18）我们之前已有描述（《血液》1985年；66：503）。它针对共价结合在α链上的纤维蛋白肽A，即针对Aα链的氨基末端片段。Y18与辣根过氧化物酶偶联，用作标记抗体。该EIA不与浓度高达800微克/毫升的纯化片段X和Y等FgDP发生反应，也不受其干扰。链激酶处理血浆产生的FgDP混合物无反应。浓度高达800微克/毫升的纤维蛋白降解产物（全血裂解物）、肝素、乙二胺四乙酸（EDTA）或草酸盐均无干扰。该EIA的出结果时间仅为45分钟。一些患者血浆产生的剂量反应曲线与EIA的校准曲线不平行。无法对这一现象作出解释。（摘要截短至250字）
