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Overview of a high-throughput pipeline for streamlining the production of recombinant proteins

Abstract:
Production of high quality protein is an essential step for both structural and functional studies. Throughput has increased in the past decade by the use of streamlined workflows with standard operating procedures and automation. In this chapter, we describe the Oxford Protein Production Facility (OPPF) pipeline for protein production, from conception, through vector construction, to expression and purification. Results from projects run in the OPPF demonstrate the value of using parallel expression screening of intracellular proteins in both E. coli and insect cells. Transient expression in Human Embryonic Kidney (HEK) cells is used exclusively for production of secreted glycoproteins. Protein purification and quality assessment are independent of the expression system and enable sample preparation to be simplified and streamlined.
Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1007/978-1-4939-9624-7_2

Authors


More by this author
Institution:
University of Oxford
Division:
MSD
Department:
NDM
Sub department:
Structural Biology
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MSD
Department:
NDM
Sub department:
Structural Biology
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MSD
Department:
NDM
Sub department:
Structural Biology
Role:
Author
ORCID:
0000-0002-3705-2993

Contributors

Role:
Editor


Publisher:
Humana Press
Host title:
High-Throughput Protein Production and Purification
Pages:
33-49
Chapter number:
2
Series:
Methods in Molecular Biology
Series number:
2025
Place of publication:
New York
Publication date:
2019-07-03
Acceptance date:
2018-11-05
DOI:
EISSN:
1940-6029
ISSN:
1064-3745
Pmid:
31267447
EISBN:
9781493996247
ISBN:
9781493996230


Language:
English
Pubs id:
pubs:1031919
UUID:
uuid:8c81bfc2-c346-4dbe-bcc1-47db8a9ccf42
Local pid:
pubs:1031919
Source identifiers:
1031919
Deposit date:
2020-01-10

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