College of Chinese Traditional Veterinary Science, Agricultural University of Hebei, Dingzhou 073000, China.
Biochem Biophys Res Commun. 2010 Mar 19;393(4):703-7. doi: 10.1016/j.bbrc.2010.02.063. Epub 2010 Feb 17.
A single-chain antibody library against Eimeria acervulina merozoites was constructed by phage display approach. Antibody-displaying phage was selected in four panning rounds against cryopreserved E. acervulina merozoites. Five clones were randomly selected from the fourth panning round, and their nucleotide sequences were aligned and compared to mouse germ-line sequences. Soluble antibody was produced in a non-suppressor Escherichia coli strain, purified by protein A affinity chromatography, and characterized by Western-blotting. Immunofluorescence assay showed localization of the produced recombinant antibody fragment on the surface E. acervulina merozoites. These resultant antibody fragments showed high specificity and binding capacity for soluble antigens and intact fixed merozoites which seems promising as diagnostic, therapeutic and/or vaccine tools against coccidiosis.
通过噬菌体展示技术构建了针对堆形艾美耳球虫裂殖子的单链抗体文库。在四轮淘选过程中,用冷冻保存的堆形艾美耳球虫裂殖子对抗体展示噬菌体进行了选择。从第四轮淘选随机选择了 5 个克隆,并对其核苷酸序列进行了比对分析,并与小鼠 germ-line 序列进行了比较。在非抑制性大肠杆菌菌株中产生了可溶性抗体,通过蛋白 A 亲和层析进行了纯化,并通过 Western-blotting 进行了表征。免疫荧光试验显示,所产生的重组抗体片段定位于堆形艾美耳球虫裂殖子表面。这些抗体片段表现出对可溶性抗原和固定完整裂殖子的高特异性和结合能力,有望成为抗球虫病的诊断、治疗和/或疫苗工具。