Semiautomated Small-Scale Purification Method for High-Throughput Expression Analysis of Recombinant Proteins.

BEVS Baculovirus E. coli High-throughput protein expression Mammalian transient expression Parallel protein purification Recombinant protein expression

Journal

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969

Informations de publication

Date de publication:
2019
Historique:
entrez: 4 7 2019
pubmed: 4 7 2019
medline: 31 3 2020
Statut: ppublish

Résumé

The expression analysis of recombinant proteins is a challenging step in any high-throughput protein production pipeline. Often multiple expression systems and a variety of expression construct designs are considered for the production of a protein of interest. There is a strong need to triage constructs rapidly and systematically. This chapter describes a semiautomated method for the simultaneous purification and characterization of proteins expressed from multiple samples of expression cultures from the E. coli, baculovirus expression vector system, and mammalian transient expression systems. This method assists in the selection of the most promising expression construct(s) or the most favorable expression condition(s) to move forward into large-scale protein production.

Identifiants

pubmed: 31267448
doi: 10.1007/978-1-4939-9624-7_3
doi:

Substances chimiques

Recombinant Proteins 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

51-68

Auteurs

Edward Kraft (E)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Yvonne Franke (Y)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Katharine Heeringa (K)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Stephanie Shriver (S)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Inna Zilberleyb (I)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Christine Kugel (C)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Trisha Dela Vega (T)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Athena Wong (A)

Department of Early Stage Cell Culture, Genentech, Inc., South San Francisco, CA, USA.

Bobby Brillantes (B)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Claudio Ciferri (C)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

George Dutina (G)

Department of Early Stage Cell Culture, Genentech, Inc., South San Francisco, CA, USA.

Grace Lee (G)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Isabelle Lehoux (I)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Zhong Rong Li (ZR)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Lee Lior-Hoffmann (L)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Jiyoung Hwang (J)

23andMe Inc., South San Francisco, CA, USA.

Chris Lonergan (C)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Lynn Martin (L)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Kyle Mortara (K)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Lananh Nguyen (L)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Jian Payandeh (J)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Andrew Perez (A)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Jun Sampang (J)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Lovejit Singh (L)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Kurt Schroeder (K)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Christine Tam (C)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Shu Ti (S)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Ye Naing Win (YN)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA.

Krista Bowman (K)

Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA. krafte1@gene.com.

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Classifications MeSH