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On-chip immunoprecipitation for protein purification

Sandison, Mairi E. and Cumming, Sarah A. and Kolch, Walter and Pitt, Andrew R. (2010) On-chip immunoprecipitation for protein purification. Lab on a Chip, 10 (20). pp. 2805-2813. ISSN 1473-0197

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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.