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| Surface Display of Functional RBD of SARS-CoV-2 in Pichia pastoris |
YU Lu1,HU Xuan1,ZHANG Xiao-juan1,2,NIU An-na1,ZHANG Xiao-peng1,**( ) |
1. Institute of Biotechnology, Academy of Military Medical Science, Beijing 100071, China
2. Institutes of Physical Science and Information Technology, Anhui University, Hefei 230601, China |
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Abstract Objective: To establish a high-throughput platform for drug discovery targeting receptor binding domain (RBD) of SARS-CoV-2, a surface display system was designed and constructed to deliver functional RBD to the surface of Pichia pastoris. Methods: Four anchor molecules were fused to RBD, and then were transformed into Pichia pastoris by using electroporation. The surface display efficiency of RBD was measured using flow cytometry, and the affinity of RBD binding to the ACE2 receptor was further determined. Results: RBD-Sed1p system exhibited the highest surface display efficiency of 70%. The binding affinity to ACE2 of RBD displayed on the cellular surface (KD=30.42 nmol/L) was close to that of RBD in solution (KD=16.00 nmol/L). Conclusion: A surface display system of RBD was successfully developed in Pichia pastoris, which can be used for high-throughput screening and evaluation of anti-COVID-19 drugs.
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Received: 23 March 2022
Published: 07 July 2022
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Corresponding Authors:
Xiao-peng ZHANG
E-mail: zxp8565@aliyun.com
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