Isolation and characterization of full-length genes encoding the anti-human CD45 antibody from the hybridoma cell line 16E8-F2

N. Q. Huy, Ta Huong Giang, N. Quan
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Abstract

Though the hybridoma technology has been widely applied in the production of monoclonal antibodies, it has existed some disadvantages including low yield and genetic instability. Therefore, an alternative approach should be taken into account. Recently, recombinant monoclonal antibody technology has emerged as the best choice to cure hybridoma related drawbacks. However, recombinant antibodies require known genes for their generation. The purpose of this study is to collect the full-length genes encoding the anti-human CD45 antibody derived from the hybridoma cell line 16E8-F2. In this research, we designed specific primer pairs to amplify the light and heavy chain genes of the antibody through the PCR method. Afterward, the genes were separately cloned into a cloning vector called pJET1.2/blunt. The generated recombinant pJET1.2 vectors will serve as the main material source to manufacture the recombinant monoclonal antibody recognizing human CD45 protein tomorrow.
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16E8-F2杂交瘤细胞系抗人CD45抗体全长基因的分离与鉴定
虽然杂交瘤技术在单克隆抗体的生产中得到了广泛的应用,但它存在着产量低、遗传不稳定等缺点。因此,应考虑另一种办法。近年来,重组单克隆抗体技术已成为治疗杂交瘤相关缺陷的最佳选择。然而,重组抗体的产生需要已知的基因。本研究的目的是从杂交瘤细胞系16E8-F2中收集编码抗人CD45抗体的全长基因。在本研究中,我们设计了特异性引物对,通过PCR方法扩增抗体的轻链和重链基因。随后,将这些基因分别克隆到名为pJET1.2/blunt的克隆载体中。生成的重组pJET1.2载体将作为主要材料来源,用于制备识别人CD45蛋白的重组单克隆抗体。
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审稿时长
8 weeks
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