Literature DB >> 30242676

Transient Gene Expression in Suspension HEK293-EBNA1 Cells.

Denis L'Abbé1, Louis Bisson1, Christian Gervais1, Eric Grazzini1, Yves Durocher2.   

Abstract

Transient gene expression in human embryo kidney 293 (HEK293) cells is an established approach for the rapid production of large amounts of recombinant proteins (r-proteins). Milligram to gram quantities of r-proteins can be typically obtained within less than 10 days following transfection. In this chapter, we describe a simple and robust transfection process of suspension-growing human embryo kidney 293 cells using two commercially available serum-free media and polyethylenimine as the transfection reagent. This chapter provides examples for the production and purification of a his-tagged recombinant protein and two monoclonal antibodies.

Entities:  

Keywords:  293-6E; Antibody; Deacylated PEI; IMAC; Polyethylenimine; Protein-A; Recombinant protein; Serum-free medium; Suspension culture; Transfection

Mesh:

Substances:

Year:  2018        PMID: 30242676     DOI: 10.1007/978-1-4939-8730-6_1

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN: 1064-3745


  7 in total

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3.  High-yield production of human Dicer by transfection of human HEK293-EBNA1 cells grown in suspension.

Authors:  Jonathan Bouvette; Dursun Nizam Korkut; Aurélien Fouillen; Soumiya Amellah; Antonio Nanci; Yves Durocher; James G Omichinski; Pascale Legault
Journal:  BMC Biotechnol       Date:  2018-12-06       Impact factor: 2.563

4.  Golden Gate assembly with a bi-directional promoter (GBid): A simple, scalable method for phage display Fab library creation.

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5.  Facile isolation of high-affinity nanobodies from synthetic libraries using CDR-swapping mutagenesis.

Authors:  Jennifer M Zupancic; Alec A Desai; Peter M Tessier
Journal:  STAR Protoc       Date:  2022-01-20

6.  Endogenous DNA 3' Blocks Are Vulnerabilities for BRCA1 and BRCA2 Deficiency and Are Reversed by the APE2 Nuclease.

Authors:  Alejandro Álvarez-Quilón; Jessica L Wojtaszek; Marie-Claude Mathieu; Tejas Patel; C Denise Appel; Nicole Hustedt; Silvia Emma Rossi; Bret D Wallace; Dheva Setiaputra; Salomé Adam; Yota Ohashi; Henrique Melo; Tiffany Cho; Christian Gervais; Ivan M Muñoz; Eric Grazzini; Jordan T F Young; John Rouse; Michael Zinda; R Scott Williams; Daniel Durocher
Journal:  Mol Cell       Date:  2020-06-08       Impact factor: 17.970

7.  The CAG promoter maintains high-level transgene expression in HEK293 cells.

Authors:  Yuanyuan Dou; Yan Lin; Tian-Yun Wang; Xiao-Yin Wang; Yan-Long Jia; Chun-Peng Zhao
Journal:  FEBS Open Bio       Date:  2020-12-03       Impact factor: 2.792

  7 in total

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