PMID- 30242687 OWN - NLM STAT- MEDLINE DCOM- 20190513 LR - 20190513 IS - 1940-6029 (Electronic) IS - 1064-3745 (Linking) VI - 1850 DP - 2018 TI - High Throughput Transfection of HEK293 Cells for Transient Protein Production. PG - 179-187 LID - 10.1007/978-1-4939-8730-6_12 [doi] AB - 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. FAU - Arena, Tia A AU - Arena TA AD - Department of Early Stage Cell Culture, Genentech Inc., South San Francisco, CA, USA. FAU - Harms, Peter D AU - Harms PD AD - Department of Late Stage Cell Culture, Genentech Inc., South San Francisco, CA, USA. FAU - Wong, Athena W AU - Wong AW AD - Department of Early Stage Cell Culture, Genentech Inc., South San Francisco, CA, USA. wong.athena@gene.com. LA - eng PT - Journal Article PL - United States TA - Methods Mol Biol JT - Methods in molecular biology (Clifton, N.J.) JID - 9214969 RN - 0 (Recombinant Proteins) SB - IM MH - Biotechnology MH - HEK293 Cells MH - High-Throughput Screening Assays/methods MH - Humans MH - Recombinant Proteins/genetics/*metabolism MH - Transfection/*methods OTO - NOTNLM OT - Automation OT - HEK293 cells OT - High throughput OT - Polyethylenimine OT - Recombinant protein production OT - Transient transfection EDAT- 2018/09/23 06:00 MHDA- 2019/05/14 06:00 CRDT- 2018/09/23 06:00 PHST- 2018/09/23 06:00 [entrez] PHST- 2018/09/23 06:00 [pubmed] PHST- 2019/05/14 06:00 [medline] AID - 10.1007/978-1-4939-8730-6_12 [doi] PST - ppublish SO - Methods Mol Biol. 2018;1850:179-187. doi: 10.1007/978-1-4939-8730-6_12.