Designing Experiments for High-Throughput Protein Expression

  • Stephen P. Chambers
  • Susanne E. Swalley
Part of the Methods in Molecular Biology book series (MIMB, volume 498)


The advent of high-throughput protein production and the vast amount of data it is capable of generat ing has created both new opportunities and problems. Automation and miniaturization allow experimen tation to be performed more efficiently, justifying the cost involved in establishing a high-throughput platform. These changes have also magnified the need for effective statistical methods to identify trends and relationships in the data. The application of quantitative management tools to this process provides the means of ensuring maximum efficiency and productivity.

Key words:

Protein expression optimization Quantitative analysis Experimental design Screening Statistics 


  1. 1.
    Bousse L, Mouradian S, Minalla A, Yee H, Williams K, Dubrow R. (2001) Protein sizing on a microchip. Anal Chem 73(6):1207–12.CrossRefPubMedGoogle Scholar
  2. 2.
    Box GEP, Hunter JS, Hunter WG. 2005 Statistics for Experimenters: Design, Innovation, and Discovery. 2nd ed. New York: Willey.Google Scholar
  3. 3.
    Chen GQ, Sun Y, Jin R, Gouaux E. (1998) Probing the d binding domain of the GluR2 receptor by proteolysis and deletion muta genesis defines domain boundaries and yields a crystallizable construct. Protein Sci 7(12):2623–30.CrossRefPubMedGoogle Scholar
  4. 4.
    Zhang Z, Smith DL. (1996) Thermal-induced unfolding domains in aldolase iden tified by amide hydrogen exchange and mass spectrometry. Protein Sci 5(7):1282–9.CrossRefPubMedGoogle Scholar
  5. 5.
    Chambers SP, Austen DA, Fulghum JR, Kim WM. (2004) High-throughput screening for soluble recombinant expressed kinases in Escherichia coli and insect cells. Protein Expr Purif 36(1):40–7.CrossRefPubMedGoogle Scholar
  6. 6.
    Studier FW, Moffatt BA. (1986) Use of bac teriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol 189(1):113–30.CrossRefPubMedGoogle Scholar
  7. 7.
    Schein CH, Noteborn MHM. (1988) For mation of soluble recombinant proteins in Escherichia coli is favored by lower growth temperature. Biotechnology 6:291–4.CrossRefGoogle Scholar
  8. 8.
    Bocanegra JA, Bejarano LA, Valdivia MM. (1997) Expression of the highly toxic centromere binding protein CENP-B in E. coli using the pET system in the absence of the inducer IPTG. Biotechniques 22(5): 798–800, 802.PubMedGoogle Scholar
  9. 9.
    Licari PB, Bailey J. (1991) Factors influ encing recombinant protein yields in insect cell-baculovirus expression systems: multiplicity of infection and intracellular protein degradation. Biotechnol Bioeng 37:238–46.CrossRefPubMedGoogle Scholar
  10. 10.
    Moore JT, Uppal A, Maley F, Maley GF. (1993) Overcoming inclusion body for mation in a high-level expression system. Protein Expr Purif 4(2):160–3.CrossRefPubMedGoogle Scholar
  11. 11.
    De Francesco R, Urbani A, Nardi MC, Tomei L, Steinkuhler C, Tramontano A. (1996) A zinc binding site in viral serine proteinases. Biochemistry 35(41):13282–7.CrossRefPubMedGoogle Scholar
  12. 12.
    Kumagai A, Dunphy WG. (1996) Puri fication and molecular cloning of Plx1, a Cdc25-regulatory kinase from Xenopus egg extracts. Science 273(5280):1377–80.CrossRefPubMedGoogle Scholar
  13. 13.
    Ying BW, Taguchi H, Kondo M, Ueda T. (2005) Co-translational involvement of the chaperonin GroEL in the folding of newly translated polypeptides. J Biol Chem 280(12):12035–40.CrossRefPubMedGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Stephen P. Chambers
    • 1
  • Susanne E. Swalley
    • 1
  1. 1.Gene ExpressionVertex PharmaceuticalsCambridgeUSA

Personalised recommendations