Cell-free systems for accelerating glycoprotein expression and biomanufacturing.

Jasmine Hershewe, Weston Kightlinger, Michael C Jewett

Journal: Journal of industrial microbiology & biotechnology 2021;47(11):977-991

PMID: 33090335

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Abstract

Protein glycosylation, the enzymatic modification of amino acid sidechains with sugar moieties, plays critical roles in cellular function, human health, and biotechnology. However, studying and producing defined glycoproteins remains challenging. Cell-free glycoprotein synthesis systems, in which protein synthesis and glycosylation are performed in crude cell extracts, offer new approaches to address these challenges. Here, we review versatile, state-of-the-art systems for biomanufacturing glycoproteins in prokaryotic and eukaryotic cell-free systems with natural and synthetic N-linked glycosylation pathways. We discuss existing challenges and future opportunities in the use of cell-free systems for the design, manufacture, and study of glycoprotein biomedicines.

Address: Department of Chemical and Biological Engineering, Northwestern University, Technological Institute E136, 2145 Sheridan Road, Evanston, IL, 60208-3120, USA.; Chemistry of Life Processes Institute, Northwestern University, 2170 Campus Drive, Evanston, IL, 60208-3120, USA.; Center for Synthetic Biology, Northwestern University, Technological Institute E136, 2145 Sheridan Road, Evanston, IL, 60208-3120, USA.; Department of Chemical and Biological Engineering, Northwestern University, Technological Institute E136, 2145 Sheridan Road, Evanston, IL, 60208-3120, USA. [email protected].; Chemistry of Life Processes Institute, Northwestern University, 2170 Campus Drive, Evanston, IL, 60208-3120, USA. [email protected].; Center for Synthetic Biology, Northwestern University, Technological Institute E136, 2145 Sheridan Road, Evanston, IL, 60208-3120, USA. [email protected].; Robert H. Lurie Comprehensive Cancer Center, Northwestern University, 676 North Saint Clair Street, Suite 1200, Chicago, IL, 60611-3068, USA. [email protected].; Simpson Querrey Institute, Northwestern University, 303 East Superior Street, Suite 11-131, Chicago, IL, 60611-2875, USA. [email protected].

DOI: 10.1007/s10295-020-02321-4

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MeSH Terms: Biotechnology, Cell-Free System, Glycoproteins, Glycosylation, Humans, Prokaryotic Cells, Protein Biosynthesis