On-chip immunoprecipitation for protein purification

Mairi E. Sandison, Sarah A. Cumming, Walter Kolch, Andrew R Pitt

Research output: Contribution to journalArticle

Abstract

Immunoprecipitation (IP) is one of the most widely used and selective techniques for protein purification. Here, a miniaturised, polymer-supported immunoprecipitation (µIP) method for the on-chip purification of proteins from complex mixtures is described. A 4 µl PDMS column functionalised with covalently bound antibodies was created and all critical aspects of the µIP protocol (antibody immobilisation, blocking of potential non-specific adsorption sites, sample incubation and washing conditions) were assessed and optimised. The optimised µIP method was used to obtain purified fractions of affinity-tagged protein from a bacterial lysate.
LanguageEnglish
Pages2805-2813
Number of pages9
JournalLab on a Chip
Volume2010
Issue number20
DOIs
Publication statusPublished - 17 Aug 2010

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Protein Array Analysis
Immunoprecipitation
Purification
Proteins
Antibodies
Blocking Antibodies
Complex Mixtures
Immobilization
Adsorption
Polymers
Washing
Broncho-Vaxom

Keywords

  • equipment design
  • equipment failure analysis
  • immunoprecipitation
  • microfluidic analytical techniques
  • proteins

Cite this

Sandison, M. E., Cumming, S. A., Kolch, W., & Pitt, A. R. (2010). On-chip immunoprecipitation for protein purification. Lab on a Chip, 2010(20), 2805-2813. https://doi.org/10.1039/c005295g
Sandison, Mairi E. ; Cumming, Sarah A. ; Kolch, Walter ; Pitt, Andrew R. / On-chip immunoprecipitation for protein purification. In: Lab on a Chip. 2010 ; Vol. 2010, No. 20. pp. 2805-2813.
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Sandison, ME, Cumming, SA, Kolch, W & Pitt, AR 2010, 'On-chip immunoprecipitation for protein purification' Lab on a Chip, vol. 2010, no. 20, pp. 2805-2813. https://doi.org/10.1039/c005295g

On-chip immunoprecipitation for protein purification. / Sandison, Mairi E.; Cumming, Sarah A.; Kolch, Walter; Pitt, Andrew R.

In: Lab on a Chip, Vol. 2010, No. 20, 17.08.2010, p. 2805-2813.

Research output: Contribution to journalArticle

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