On-chip immunoprecipitation for protein purification.
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.
Sandison
Mairi E
ME
Integrative and Systems Biology, FBLS, University of Glasgow, Glasgow, G12 8QQ, UK. m.sandison@bio.gla.ac.uk
Cumming
Sarah A
SA
Kolch
Walter
W
Pitt
Andrew R
AR
D004867:Equipment Design; D019544:Equipment Failure Analysis; D047468:Immunoprecipitation / Q000295:instrumentation*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D011506:Proteins / Q000302:isolation & purification*
D011506:Proteins