Keywords

cell-free protein synthesis (CFPS), linear DNA expression templates (LET), high-throughput protein screening

Abstract

A rapid, versatile method of protein expression and screening can greatly facilitate the future development of therapeutic biologics, proteomic drug targets and biocatalysts. An attractive candidate is cell-free protein synthesis (CFPS), a cell-lysate-based in vitro expression system, which can utilize linear DNA as expression templates, bypassing time-consuming cloning steps of plasmid-based methods. Traditionally, such linear DNA expression templates (LET) have been vulnerable to degradation by nucleases present in the cell lysate, leading to lower yields. This challenge has been significantly addressed in the recent past, propelling LET-based CFPS as a useful tool for studying, screening and engineering proteins in a high-throughput manner. Currently, LET-based CFPS has promise in fields such as functional proteomics, protein microarrays, and the optimization of complex biological systems.

Original Publication Citation

Song-Min Schinn, Andrew Broadbent, William T. Bradley, Bradley C. Bundy, Protein synthesis directly from PCR: progress and applications of cell-free protein synthesis with linear DNA, New Biotechnology, Volume 33, Issue 4, 2016, Pages 480-487, ISSN 1871-6784, https://doi.org/10.1016/j.nbt.2016.04.002.

Document Type

Peer-Reviewed Article

Publication Date

2016-04-13

Publisher

New Biotechnology

Language

English

College

Ira A. Fulton College of Engineering

Department

Chemical Engineering

University Standing at Time of Publication

Full Professor

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