Li Hao, Liu Litong, Kang Xinyi, Chen Chuan-Wei, Wang Mengran, Fu Shaoqin, Zhou Qingtong, Zhao Bo, Yang Dehua, Wang Ming-Wei
Research Center for Deepsea Bioresources, Sanya 572025, China.
Shanghai Institute of Infectious Disease and Biosecurity, Fudan University, Shanghai 200032, China.
Acta Pharm Sin B. 2025 Apr;15(4):1912-1921. doi: 10.1016/j.apsb.2025.02.012. Epub 2025 Feb 18.
Fifty whitespotted bamboo sharks () of both sexes were used to establish a large capacity variable domain of the new antigen receptor (VNAR) library with a total capacity of over 10 colony-forming units (CFU). It was applied to screen VNARs against human serum albumin (HSA) and human transcription factor EB (TFEB), respectively. Meanwhile, VNAR libraries specific to HSA and TFEB with capacities above 10 CFU were obtained following conventional immunization. These two approaches were systematically studied in terms of VNAR yield and composition. By comparing the VNAR sequences obtained from naïve and antigen-immunized libraries, we found that the complementary-determining region 3 (CDR3) of the former differs in composition from that of the latter. It shares a higher degree of homology with the naïve library. Meanwhile, the binding efficiency assessed by ELISA is also different between the naïve and antigen-immunized libraries. The binding of VNARs from the TFEB-immunized library appeared to surpass that observed with the naïve libraries, whereas the performance of VNARs from the HSA-immunized library indicated that both the immunized and naïve libraries for HSA had positive binding responses in polyclonal and monoclonal ELISA. The results are useful to develop novel diagnostic and therapeutic products based on shark VNARs.
使用50条雌雄皆有的白点竹鲨来构建一个新抗原受体（VNAR）文库的大容量可变区，其总容量超过10个集落形成单位（CFU）。该文库分别用于筛选针对人血清白蛋白（HSA）和人转录因子EB（TFEB）的VNAR。同时，通过传统免疫获得了针对HSA和TFEB的容量超过10 CFU的特异性VNAR文库。从VNAR产量和组成方面对这两种方法进行了系统研究。通过比较从未经免疫的文库和抗原免疫文库中获得的VNAR序列，我们发现前者的互补决定区3（CDR3）在组成上与后者不同。它与未经免疫的文库具有更高的同源性。同时，通过酶联免疫吸附测定（ELISA）评估的结合效率在未经免疫的文库和抗原免疫文库之间也有所不同。来自TFEB免疫文库的VNAR的结合似乎超过了从未经免疫的文库中观察到的结合，而来自HSA免疫文库的VNAR的表现表明，在多克隆和单克隆ELISA中，HSA的免疫文库和未经免疫的文库都有阳性结合反应。这些结果有助于开发基于鲨鱼VNAR的新型诊断和治疗产品。
