On-chip immunoprecipitation for protein purification

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

doi:10.1039/c005295g

On-chip immunoprecipitation for protein purification

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

Analytical, Measurements and Physical Chemistry

Research output: Contribution to journal › Article › peer-review

Abstract

Immunoprecipitation (IP) is one of the most widely used and selective techniques for protein purification. Here, a miniaturised, polymer-supported immunoprecipitation (IP) method for the on-chip purification of proteins from complex mixtures is described. A 4 l PDMS column functionalised with covalently bound antibodies was created and all critical aspects of the IP protocol (antibody immobilisation, blocking of potential non-specific adsorption sites, sample incubation and washing conditions) were assessed and optimised. The optimised IP method was used to obtain purified fractions of affinity-tagged protein from a bacterial lysate.

Original language	English
Pages (from-to)	2805-2813
Number of pages	9
Journal	Lab on a Chip
Volume	10
Issue number	20
DOIs	https://doi.org/10.1039/c005295g
Publication status	Published - 21 Oct 2010

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10.1039/c005295g

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AU - Kolch, Walter

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AB - Immunoprecipitation (IP) is one of the most widely used and selective techniques for protein purification. Here, a miniaturised, polymer-supported immunoprecipitation (IP) method for the on-chip purification of proteins from complex mixtures is described. A 4 l PDMS column functionalised with covalently bound antibodies was created and all critical aspects of the IP protocol (antibody immobilisation, blocking of potential non-specific adsorption sites, sample incubation and washing conditions) were assessed and optimised. The optimised IP method was used to obtain purified fractions of affinity-tagged protein from a bacterial lysate.

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On-chip immunoprecipitation for protein purification

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