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抗人纤维蛋白单克隆抗体:与其他动物纤维蛋白的相互作用

Monoclonal antibodies to human fibrin: interaction with other animal fibrins.

作者信息

Edgell T, McEvoy F, Webbon P, Gaffney P J

机构信息

Royal Veterinary College (University of London), Hertfordshire, UK.

出版信息

Thromb Haemost. 1996 Apr;75(4):595-9.

PMID:8743185
Abstract

Four monoclonal antibodies have previously been raised in our laboratory for possible use in thrombus imaging and the targeting of thrombolytic agents. These antibodies were raised to various human fibrin-related immunogens and each antibody had been selected for its specificity towards fibrin and not fibrinogen. To study further these antibodies in animal circulation models both in vivo and in vitro, their selectivity towards human fibrin as opposed to other animal fibrins was examined. In this study dissociation constants for each antibody with each of six species fibrins (human, baboon, pig, dog, sheep and rabbit) were estimated using both fibrin clots and monolayers. Some limited data were also obtained with Sepharose-fibrin. Of the antibodies two (denoted 12B3B10 and 12B3A11) are seen to bind almost exclusively to human fibrin with dissociation constants of about 8 x 10(-10) M using fibrin clots and monolayers. These same two mabs bound to baboon fibrin with a dissociation constant of 2 x 10(-9) M, while neither displayed significant levels of binding to the fibrins from dog, pig, sheep and rabbit. The other two antibodies investigated (1H10 and 5F3) were found to bind well to fibrins of human, baboon, pig and dog, with dissociation constants in the range of 1.4-4.2 x 10(-9) M. However neither 1H10 nor 5F3 displayed significant recognition of sheep and rabbit fibrins. Both 1H10 and 5F3 were also found by means of competitive ELISA's to retain their selectivity to baboon, dog and pig fibrins in the presence of their respective fibrinogens.

摘要

此前在我们实验室制备了四种单克隆抗体,可能用于血栓成像以及溶栓剂的靶向治疗。这些抗体是针对各种与人类纤维蛋白相关的免疫原产生的,并且每种抗体都因其对纤维蛋白而非纤维蛋白原的特异性而被挑选出来。为了在体内和体外动物循环模型中进一步研究这些抗体,检测了它们对人类纤维蛋白相对于其他动物纤维蛋白的选择性。在本研究中,使用纤维蛋白凝块和单层膜估算了每种抗体与六种物种(人类、狒狒、猪、狗、绵羊和兔子)纤维蛋白的解离常数。还通过琼脂糖 - 纤维蛋白获得了一些有限的数据。在这些抗体中,有两种(分别标记为12B3B10和12B3A11)几乎只与人类纤维蛋白结合,使用纤维蛋白凝块和单层膜时解离常数约为8×10⁻¹⁰ M。这两种单克隆抗体与狒狒纤维蛋白的解离常数为2×10⁻⁹ M,而它们与狗、猪、绵羊和兔子的纤维蛋白均未表现出显著的结合水平。研究的另外两种抗体(1H10和5F3)被发现与人类、狒狒、猪和狗的纤维蛋白结合良好,解离常数在1.4 - 4.2×10⁻⁹ M范围内。然而,1H10和5F3对绵羊和兔子的纤维蛋白均未表现出显著的识别能力。通过竞争性酶联免疫吸附测定法还发现,在各自纤维蛋白原存在的情况下,1H10和5F3对狒狒、狗和猪的纤维蛋白仍保持其选择性。

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