摘要
Fifty whitespotted bamboo sharks(Chiloscyllium playgiosum)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^(9) 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^(8) 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 naı¨ve 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 naı¨ve library.Meanwhile,the binding efficiency assessed by ELISA is also different between the naı¨ve and antigen-immunized libraries.The binding of VNARs from the TFEB-immunized library appeared to surpass that observed with the naı¨ve libraries,whereas the performance of VNARs from the HSA-immunized library indicated that both the immunized and naı¨ve 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.
基金
supported by research grants from Sanya Yazhou Bay Science&Technology City Administration(SKJC-2022-01-002 to Ming-Wei Wang and SKJC-2024-03-001 to Hao Li,China)
Special Project for the Promotion of Deep-Sea Technology Industry,Hainan Deep-Sea Technology Innovation Center(DSTIC-CYCJ-2023012 to Ming-Wei Wang,China).
