Biotechnol J. 2009 Jan;4(1):51-7. doi: 10.1002/biot.200800183.

High-throughput protein production--lessons from scaling up from 10 to 288 recombinant proteins per week.

Tegel H, Steen J, Konrad A, Nikdin H, Pettersson K, Stenvall M, Tourle S, Wrethagen U, Xu L, Yderland L, Uhlén M, Hober S, Ottosson J.

Source

School of Biotechnology, Royal Institute of Technology, Stockholm, Sweden.

Abstract

The demand for high-throughput recombinant protein production has markedly increased with the increased activity in the field of proteomics. Within the Human Protein Atlas project recombinantly produced human protein fragments are used for antibody production. Here we describe how the protein expression and purification protocol has been optimized in the project to allow for handling of nearly 300 different proteins per week. The number of manual handling steps has been significantly reduced (from 18 to 9) and the protein purification has been completely automated.

PMID:: 19039781; [PubMed - indexed for MEDLINE]

Publication Types, MeSH Terms, Substances

Publication Types

Research Support, Non-U.S. Gov't

MeSH Terms

Substances

LinkOut - more resources

Full Text Sources

Supplemental Content

Save items

Related citations in PubMed

Automated 96-well purification of hexahistidine-tagged recombinant proteins on MagneHis Ni(2)+-particles. [Methods Mol Biol. 2009]
Automated 96-well purification of hexahistidine-tagged recombinant proteins on MagneHis Ni(2)+-particles.
Lin CT, Moore PA, Kery V. Methods Mol Biol. 2009; 498:129-41.
Increased levels of recombinant human proteins with the Escherichia coli strain Rosetta(DE3). [Protein Expr Purif. 2010]
Increased levels of recombinant human proteins with the Escherichia coli strain Rosetta(DE3).
Tegel H, Tourle S, Ottosson J, Persson A. Protein Expr Purif. 2010 Feb; 69(2):159-67. Epub 2009 Sep 4.
Expression screening, protein purification and NMR analysis of human protein domains for structural genomics. [J Struct Funct Genomics. 2004]
Expression screening, protein purification and NMR analysis of human protein domains for structural genomics.
Folkers GE, van Buuren BN, Kaptein R. J Struct Funct Genomics. 2004; 5(1-2):119-31.
Review Human protein atlas and the use of microarray technologies. [Curr Opin Biotechnol. 2008]
Review Human protein atlas and the use of microarray technologies.
Hober S, Uhlén M. Curr Opin Biotechnol. 2008 Feb; 19(1):30-5. Epub 2008 Jan 9.
Review High-throughput automated platform for nuclear magnetic resonance-based structural proteomics. [Expert Rev Proteomics. 2005]
Review High-throughput automated platform for nuclear magnetic resonance-based structural proteomics.
Vinarov DA, Markley JL. Expert Rev Proteomics. 2005 Jan; 2(1):49-55.

See reviews... See all...

Cited by 1 PubMed Central article

The optimization of in vitro high-throughput chemical lysis of Escherichia coli. Application to ACP domain of the polyketide synthase ppsC from Mycobacterium tuberculosis. [J Struct Funct Genomics. 2010]
The optimization of in vitro high-throughput chemical lysis of Escherichia coli. Application to ACP domain of the polyketide synthase ppsC from Mycobacterium tuberculosis.
Listwan P, Pédelacq JD, Lockard M, Bell C, Terwilliger TC, Waldo GS. J Struct Funct Genomics. 2010 Mar; 11(1):41-9. Epub 2010 Jan 13.

Related information

Related Citations
Calculated set of PubMed citations closely related to the selected article(s) retrieved using a word weight algorithm. Related articles are displayed in ranked order from most to least relevant, with the “linked from” citation displayed first.
Substance (MeSH Keyword)
PubChem chemical substance (submitted) records that are classified under the same Medical Subject Headings (MeSH) controlled vocabulary as the current articles.
Cited in PMC
Full-text articles in the PubMed Central Database that cite the current articles.

Recent activity

Clear Turn Off Turn On

High-throughput protein production--lessons from scaling up from 10 to 288 recom...
High-throughput protein production--lessons from scaling up from 10 to 288 recombinant proteins per week.
Biotechnol J. 2009 Jan ;4(1):51-7. doi: 10.1002/biot.200800183.

PubMed

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

PubMed

Result Filters

Display Settings:

Send to: