黑色素瘤细胞单域抗体文库的构建及鉴定  

Construction and Identification of a Single Domain Antibody Library of Melanoma Cells

作　　者：姬凯元[1,2] 齐淑慧 刘博 蒋书东 范瑞文[1] 董常生[1] JI Kai-Yuan;QI Shu-Hui;LIU Bo;JIANG Shu-Dong;FAN Rui-Wen;DONG Chang-Sheng(Alpaca Bioengineering Laboratory,Shanxi Agricultural University,Shanxi,Taigu 030801,China;Anhui Key Laboratory of Genetic Resources Protection and Biological Breeding for Livestock and Poultry,Anhui Agricultural University,Hefei 230036,China)

机构地区：[1]山西农业大学羊驼生物工程实验室,山西太谷030801 [2]安徽农业大学畜禽遗传资源保护与生物育种安徽省重点实验室,合肥230036

出　　处：《中国生物化学与分子生物学报》2019年第11期1294-1302,共9页Chinese Journal of Biochemistry and Molecular Biology

基　　金：山西省青年三晋学者和2018年安徽农业大学校青年基金(No.03087060)资助~~

摘　　要：黑色素瘤作为一种多发于皮肤部位的恶性肿瘤,严重危害着动物和人们的健康。与传统抗体比较,单域抗体具有结构简单、分子量小、免疫原性弱等特点,使其在疾病的诊断及治疗方面具有广阔的应用空间。本研究以B16-F10细胞蛋白质为研究对象,通过反复冻融与超声破碎相结合的方式获得B16-F10蛋白质作为抗原,免疫成年雄性羊驼。采用噬菌体单域抗体展示技术,构建了质量优良的B16-F10细胞蛋白质单域抗体免疫文库,其库容为5.76×1010,VHH重组率为96%,文库丰度为3.00×1010个/mL。该结果为研究黑色素瘤的生物学特性提供了新思路,同时也为后续筛选B16-F10单域抗体奠定了基础。Melanoma,known as a kind of skin cancer,is a serious threat to the health of animals and human.Compared with traditional antibodies,single domain antibodies have wide applications due to their simple structure,small molecular weight and weak immunogenicity.In this study,the B16-F10 protein was selected as the research object,and B16-F10 protein was obtained as the antigen through the combination of repeated freeze-thaw and ultrasonic crushing to immunize adult male alpaca.Peripheral venous blood was collected,lymphocytes were isolated,and VHH fragments encoding the nano-antibody were synthesized.The phage display technology was used to construct an immune library with a capacity of 5.76×1010,a VHH recombination rate of 96%and a library abundance of 3.00×1010/mL for B16-F10 single domain antibody.These results provide a new idea for the study of the biological characteristics of melanoma and lay a foundation for the subsequent screening of B16-F10 single domain antibodies.

关 键 词：黑色素瘤 细胞蛋白质B16-F10 羊驼 单域抗体 CDNA文库 

分 类 号：S852[农业科学—基础兽医学]