High Throughput Transfection of HEK293 Cells for Transient Protein Production

Tia A Arena; Peter D Harms; Athena W Wong

doi:10.1007/978-1-4939-8730-6_12

High Throughput Transfection of HEK293 Cells for Transient Protein Production

Methods Mol Biol. 2018:1850:179-187. doi: 10.1007/978-1-4939-8730-6_12.

Authors

Tia A Arena¹, Peter D Harms², Athena W Wong³

Affiliations

¹ Department of Early Stage Cell Culture, Genentech Inc., South San Francisco, CA, USA.
² Department of Late Stage Cell Culture, Genentech Inc., South San Francisco, CA, USA.
³ Department of Early Stage Cell Culture, Genentech Inc., South San Francisco, CA, USA. wong.athena@gene.com.

PMID: 30242687
DOI: 10.1007/978-1-4939-8730-6_12

Abstract

Transient transfection of mammalian cells is used in the biotechnology industry to quickly supply recombinant protein for research and large molecule drug development. Here, we describe a method for high throughput transient transfection of Human Embryonic Kidney 293 (HEK293) cells in 30 mL tubespins using polyethylenimine (PEI) as a transfection reagent. An automated liquid handler can be used to perform pipetting steps for transfecting batches of 96 tubespins, and septa in the tubespin caps allow for rapid processing without decapping. The addition of valproic acid (VPA) to transfection cultures enhances recombinant protein production. The thawing and passaging operations for HEK293 cultures to source the transient transfections are also described.

Keywords: Automation; HEK293 cells; High throughput; Polyethylenimine; Recombinant protein production; Transient transfection.

MeSH terms

Biotechnology
HEK293 Cells
High-Throughput Screening Assays / methods
Humans
Recombinant Proteins / genetics
Recombinant Proteins / metabolism*
Transfection / methods*

Substances

Recombinant Proteins