Quantification of Interlaboratory Cell-Free Protein Synthesis Variability

ACS Synthetic Biology
Stephanie D ColeMatthew W Lux

Abstract

Cell-free protein synthesis (CFPS) platforms, once primarily a research tool to produce difficult to express proteins, are increasingly being pursued by the synthetic biology community for applications including biomanufacturing, rapid screening systems, and field-ready sensors. While consistency within individual studies is apparent in the literature, challenges with reproducing results between laboratories, or even between individuals within a laboratory, are discussed openly by practitioners. As the field continues to grow and move toward applications, a quantitative understanding of expected variability for CFPS and the relative contribution of underlying sources will become increasingly important. Here we offer the first quantitative assessment of interlaboratory variability in CFPS. Three laboratories implemented a single CFPS protocol and performed a series of exchanges, both of material and personnel, designed to quantify relative contributions to variability associated with the site, operator, cell extract preparation, and supplemental reagent preparation. We found that materials prepared at each laboratory, exchanged pairwise, and tested at each site resulted in 40.3% coefficient of variation compared to 7.64% for a s...Continue Reading

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Citations

Nov 30, 2019·Nature Reviews. Genetics·Adam D SilvermanMichael C Jewett
Dec 28, 2019·International Journal of Molecular Sciences·Jascha RolfKatrin Rosenthal
Aug 15, 2020·Frontiers in Bioengineering and Biotechnology·Nadanai Laohakunakorn
Jul 16, 2020·Biotechnology Journal·Liyuan ZhangYuan Lu
Apr 9, 2020·Frontiers in Bioengineering and Biotechnology·Nadanai LaohakunakornSebastian J Maerkl
Oct 28, 2020·Current Protocols in Protein Science·Rochelle AwKaren M Polizzi
Jan 12, 2021·RNA Biology·Yaroslav ChushakNancy Kelley-Loughnane
Nov 17, 2020·Frontiers in Bioengineering and Biotechnology·Jan MüllerRalf Takors
Mar 4, 2021·Nature Communications·Sierra M Brooks, Hal S Alper
Aug 11, 2020·Synthetic and Systems Biotechnology·Stephanie D ColeMatthew W Lux
Apr 13, 2021·ACS Synthetic Biology·Steven M BlumMatthew W Lux
Apr 24, 2021·Nature Communications·Jasmine M HersheweMichael C Jewett
Jul 3, 2021·Life·Conary MeyerCheemeng Tan
Jul 11, 2020·ACS Synthetic Biology·Marilyn S LeeMatthew W Lux

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