【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。更多还原

【Abstract】 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.更多还原