Method for recombinant expression of human CD20 by using E.coli cell-free protein expression system

• Main Authors: LIN SEN, LIU XIAYUE, XU HANYU, WANG FANG, TANG YAJIE, WANG YAO, CAI HAIYING, WANG ZAIHUA, LU CAITING, JIANG LELE, CHEN ZHILING, WU YINRONG, TANG HONGFENG
• Format: Patent
• Language: Chinese; English
• Published: 22.03.2022
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention discloses a method for recombinant expression of human CD20 by using an E.coli cell-free protein expression system. The method comprises the following steps: (1) amplifying coding DNA of human CD20 protein; and (2) adding the amplification product obtained in the step (1) into an E.coli cell-free protein expression system to synthesize the human CD20 protein. An E.coli cell-free protein expression system is adopted to recombine and express the human CD20 protein outside cells, the human CD20 comprises full-length human CD20 and homologous protein of the human CD20 (the similarity with the amino acid sequence of the full-length human CD20 exceeds 30%), the production cycle of the human CD20 is greatly shortened, the production cost is reduced, and the production efficiency is improved; in addition, the protein expression process is high in controllability, and the yield of human CD20 can reach 2 mg/mL or above. 本发明公开了一种利用E.coli无细胞蛋白表达系统重组表达人CD20的方法，该方法包括以下步骤：(1)扩增人CD20蛋白的编码DNA；(2)将步骤(1)获得的扩增产物加入到