Streamlining the preparation of “endotoxin-free” ClearColi cell extract with autoinduction media for cell-free protein synthesis of the therapeutic protein crisantaspase

作     者：J.Porter Hunt Emily Long Zhao Mehran Soltani Madison Frei J.Andrew D.Nelson Bradley C.Bundy 

作者机构：Department of Chemical EngineeringBrigham Young UniversityProvoUTUSA Department of Chemical EngineeringBrigham Young University330N Engineering BuildingProvoUT84602USA 

出 版 物：《Synthetic and Systems Biotechnology》 (合成和系统生物技术（英文）)

年 卷 期：2019年第4卷第4期

页      面：220-224页

核心收录：

学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学] 

基　　金：This work was supported by the National Science Foundation CBET Division CAREER Award(grant number:1254148) the Utah NASA Space Grant Consortium and by the Simmons Center for Cancer Research at Brigham Young University 

主　　题：Auto-induction Autoinduction Cell-free protein synthesis CFPS Endotoxin-free Crisantaspase Clearcoli 

摘      要：An“endotoxin-free***-based cell-free protein synthesis system has been reported to produce therapeutic proteins rapidly and ***,preparation of the most complex CFPS reagent–the cell extract–remains time-consuming and labor-intensive because of the relatively slow growth kinetics of the endotoxin-free ClearColiTMBL21(DE3)*** we report a streamlined procedure for preparing *** cell extract from ClearColi™using auto-induction *** this work,the term auto-induction describes cell culture media which eliminates the need for manual induction of protein *** Clearcoli™cells in autoinduction media significantly reduces the hands-on time required during extract preparation,and the resulting“endotoxinfreecell extract maintained the same cell-free protein synthesis capability as extract produced with traditional induction as demonstrated by the high-yield expression of crisantaspase,an FDA approved leukemia *** is anticipated that this work will lower the barrier for researchers to enter the field and use this technology as the method to produce endotoxin-free ***-based extract for CFPS.