Literature DB >> 16013618

Production of milligram quantities of affinity tagged-proteins using automated multistep chromatographic purification.

Rama Bhikhabhai1, Anna Sjöberg, Lotta Hedkvist, Markus Galin, Pia Liljedahl, Tuomo Frigård, Niklas Pettersson, Mats Nilsson, Jill A Sigrell-Simon, Christine Markeland-Johansson.   

Abstract

A new chromatography system, AKTAxpress (GE Healthcare, Amersham Biosciences, Uppsala, Sweden) has been designed to meet the demand for high-throughput purification of proteins in structural genomics and drug discovery. The system offers a number of automated multistep purification protocols for affinity-tagged proteins. All protocols start with affinity chromatography followed by combinations of desalting, ion exchange chromatography and gel filtration. As an option, tag removal can be included in the purification protocols. Up to 16 proteins can be purified per day and the yield can be as high as 50 mg of each protein at > 90% purity. To highlight the versatility of the system, this paper presents several case studies; purifications of hexahistidine- and glutathione S-transferase-tagged proteins using different protocols, automated on-column tag cleavage and optimization studies for a hexahistidine-tagged kinase.

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Year:  2005        PMID: 16013618     DOI: 10.1016/j.chroma.2005.05.026

Source DB:  PubMed          Journal:  J Chromatogr A        ISSN: 0021-9673            Impact factor:   4.759


  7 in total

Review 1.  Chapter 3. High-throughput protein purification for x-ray crystallography and NMR.

Authors:  Youngchang Kim; Lance Bigelow; Maria Borovilos; Irina Dementieva; Erika Duggan; William Eschenfeldt; Catherine Hatzos; Grazyna Joachimiak; Hui Li; Natalia Maltseva; Rory Mulligan; Pearl Quartey; Alicia Sather; Lucy Stols; Lour Volkart; Ruiying Wu; Min Zhou; Andrzej Joachimiak
Journal:  Adv Protein Chem Struct Biol       Date:  2009-02-26       Impact factor: 3.507

2.  Surface interactome in Streptococcus pyogenes.

Authors:  Cesira L Galeotti; Elia Bove; Alfredo Pezzicoli; Renzo Nogarotto; Nathalie Norais; Silvia Pileri; Barbara Lelli; Fabiana Falugi; Sergio Balloni; Vittorio Tedde; Emiliano Chiarot; Mauro Bombaci; Marco Soriani; Luisa Bracci; Guido Grandi; Renata Grifantini
Journal:  Mol Cell Proteomics       Date:  2011-12-22       Impact factor: 5.911

3.  Fully automated protein purification.

Authors:  DeMarco V Camper; Ronald E Viola
Journal:  Anal Biochem       Date:  2009-07-28       Impact factor: 3.365

4.  Streamlined, automated protocols for the production of milligram quantities of untagged recombinant human cyclophilin-A (hCypA) and untagged human proliferating cell nuclear antigen (hPCNA) using AKTAxpress.

Authors:  Cornelia Ludwig; Martin A Wear; Malcolm D Walkinshaw
Journal:  Protein Expr Purif       Date:  2009-12-06       Impact factor: 1.650

Review 5.  Protein production from the structural genomics perspective: achievements and future needs.

Authors:  Steven C Almo; Scott J Garforth; Brandan S Hillerich; James D Love; Ronald D Seidel; Stephen K Burley
Journal:  Curr Opin Struct Biol       Date:  2013-05-01       Impact factor: 6.809

6.  A Streamlined, Automated Protocol for the Production of Milligram Quantities of Untagged Recombinant Rat Lactate Dehydrogenase A Using ÄKTAxpressTM.

Authors:  Matthew W Nowicki; Elizabeth A Blackburn; Iain W McNae; Martin A Wear
Journal:  PLoS One       Date:  2015-12-30       Impact factor: 3.240

7.  REVOLVER: A low-cost automated protein purifier based on parallel preparative gravity column workflows.

Authors:  Patrick Diep; Jose L Cadavid; Alexander F Yakunin; Alison P McGuigan; Radhakrishnan Mahadevan
Journal:  HardwareX       Date:  2022-03-11
  7 in total

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