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牛白细胞介素-4特异性单克隆抗体的制备与鉴定

Production and characterisation of monoclonal antibodies specific for bovine interleukin-4.

作者信息

Weynants V, Gilson D, Furger A, Collins R A, Mertens P, De Bolle X, Heussler V T, Roditi I, Howard C J, Dobbelaere A E, Letesson J J

机构信息

Unité d'Immunologie-Microbiologie, Facultés Universitaires Notre-Dame de la Paix, Namur, Belgium.

出版信息

Vet Immunol Immunopathol. 1998 Nov 24;66(2):99-112. doi: 10.1016/s0165-2427(98)00184-6.

Abstract

Genetic immunisation is a simple method for producing polyclonal antibodies in mice. By this method, we produced antibodies against bovine interleukin-4 (BoIL-4). After a final injection with a recombinant BoIL-4 protein, nine stable hybridoma cell lines were established which secreted monoclonal antibodies (MAbs) against this cytokine. Specific binding of each of the MAbs to recombinant BoIL-4 produced by Escherichia coli, baculovirus, and Trypanosoma brucei was demonstrated in an indirect ELISA and/or in Western blotting. These MAbs recognise the same antigenic region localised in the first 47 amino acids of the mature protein. None of them was able to neutralise the biological activity of the BoIL-4 under the conditions tested but one allowed the detection of BoIL-4 by flow cytometry.

摘要

基因免疫是在小鼠体内产生多克隆抗体的一种简单方法。通过这种方法,我们制备了抗牛白细胞介素-4(BoIL-4)的抗体。在用重组BoIL-4蛋白进行最后一次注射后,建立了9个稳定的杂交瘤细胞系,它们分泌针对这种细胞因子的单克隆抗体(MAb)。在间接ELISA和/或蛋白质免疫印迹中证实了每种单克隆抗体与大肠杆菌、杆状病毒和布氏锥虫产生的重组BoIL-4的特异性结合。这些单克隆抗体识别位于成熟蛋白前47个氨基酸中的相同抗原区域。在所测试的条件下,它们均不能中和BoIL-4的生物活性,但有一种单克隆抗体可通过流式细胞术检测BoIL-4。

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