检测尖角艾美耳球虫表面抗原的重组ScFv抗体的制备与鉴定。

J. K. Kim, W. Min, H. Lillehoj, S. Kim, E. Sohn, K. Song, J. Y. Han
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引用次数: 15

摘要

在我们之前的尝试中,我们产生了两种鸡b细胞杂杂瘤,5D11和2-1,以更准确地反映天然禽类体液免疫反应,以产生抗球虫寄生虫的单克隆抗体(mab)。虽然这两种细胞系都能分泌与尖角艾美球虫孢子子反应的抗体,但它们的产量太低,无法进行有意义的体内研究。为了解决这个问题,我们从杂交瘤5D11和2-1的V(H)和V(L)基因中制备了四种单链可变片段(scFv)抗体。这些重组抗体在大肠杆菌中表达并纯化至均匀性,其浓度为5-6 mg/L。其中三种抗体对艾美耳球虫表面抗原表现出与天然单克隆抗体相当的抗原结合特异性。对杂交瘤5D11和2-1的V(L)基因和基因组DNA进行核苷酸序列分析,发现了与V(lambda)假基因转换的痕迹。这些重组scFv抗体将被证明有助于进一步表征天然艾美耳球虫表面抗原作为潜在的候选疫苗。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Generation and characterization of recombinant ScFv antibodies detecting Eimeria acervulina surface antigens.
In our previous attempt to generate monoclonal antibodies (MAbs) against coccidia parasites that more accurately reflect the natural avian humoral immune response, we produced two chicken B-cell hybridomas, 5D11 and 2-1. While both cell lines secreted antibodies reactive with sporozoites of Eimeria acervulina, they were produced in yields too low to conduct meaningful in vivo studies. To circumvent this problem, we produced four single chain variable fragment (scFv) antibodies from the V(H) and V(L) genes of hybridomas 5D11 and 2-1. The concentration of these recombinant antibodies expressed in E. coli and purified to homogeneity was 5-6 mg/L. Three of the antibodies exhibited antigen binding specificity to Eimeria surface antigens equivalent to that of the native MAbs. Nucleotide sequence analysis of the V(L) genes from hybridomas 5D11 and 2-1 and genomic DNA revealed vestiges of gene conversion with V(lambda) pseudogenes. These recombinant scFv antibodies will prove useful for further characterization of natural Eimeria surface antigens as potential vaccine candidates.
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来源期刊
Hybridoma
Hybridoma 医学-免疫学
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