Construction and application of a large capacity VNAR library from the whitespotted bamboo shark (<i>Chiloscyllium playgiosum</i>).

Hao LI; Litong LIU; Xinyi KANG; Chuan-Wei CHEN; Mengran WANG; Shaoqin FU; Qingtong ZHOU; Bo ZHAO; Dehua YANG; Ming-Wei WANG

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Construction and application of a large capacity VNAR library from the whitespotted bamboo shark (Chiloscyllium playgiosum).

Author: Hao LI ¹ ; Litong LIU ² ; Xinyi KANG ³ ; Chuan-Wei CHEN ¹ ; Mengran WANG ⁴ ; Shaoqin FU ⁵ ; Qingtong ZHOU ¹ ; Bo ZHAO ⁴ ; Dehua YANG ¹ ; Ming-Wei WANG ¹
Author Information

1. Research Center for Deepsea Bioresources, Sanya 572025, China.
2. Shanghai Institute of Infectious Disease and Biosecurity, Fudan University, Shanghai 200032, China.
3. Department of Pharmacology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China.
4. School of Pharmaceutical Sciences, Shanghai Jiao Tong University, Shanghai 200240, China.
5. Yazhou Bay Agricultural and Fishery Development Co., Ltd., Sanya 572000, China.
Publication Type:Journal Article
Keywords: Chiloscyllium playgiosum; HSA; Immunization; Phage display; Shark; TFEB; VNAR
From: Acta Pharmaceutica Sinica B 2025;15(4):1912-1921
CountryChina
Language:English
Abstract: 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 10⁹ 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 10⁸ 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.