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

Autor: Kraft E; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Franke Y; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Heeringa K; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Shriver S; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Zilberleyb I; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Kugel C; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Dela Vega T; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Wong A; Department of Early Stage Cell Culture, Genentech, Inc., South San Francisco, CA, USA., Brillantes B; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Ciferri C; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Dutina G; Department of Early Stage Cell Culture, Genentech, Inc., South San Francisco, CA, USA., Lee G; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Lehoux I; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Li ZR; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Lior-Hoffmann L; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Hwang J; 23andMe Inc., South San Francisco, CA, USA., Lonergan C; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Martin L; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Mortara K; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Nguyen L; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Payandeh J; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Perez A; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Sampang J; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Singh L; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Schroeder K; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Tam C; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Ti S; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Win YN; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA., Bowman K; Department of Biomolecular Resources, Genentech, Inc., South San Francisco, CA, USA. krafte1@gene.com.
Jazyk: angličtina
Zdroj: Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2019; Vol. 2025, pp. 51-68.
DOI: 10.1007/978-1-4939-9624-7_3
Abstrakt: 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.
Databáze: MEDLINE