ANYL 30 |
| IMAC is used as a tool for protein/peptide purifications in a wide variety of applications. Because fractions collected from IMAC cartridges usually contain impurities, there is a need for a high resolution IMAC-HPLC column. To achieve this, we used living/controlled radical polymerization and polymer chain collapse to engineer the surface of 10um, nonporous, polymeric beads with isolated, metal-containing nanoparticles that act as IMAC interaction sites. Surface-bound nanoscale particles were visible in transmission electron microscope (TEM) images of bead cross-sections. TEM was also used to visualize individual ferritin molecules interacting with nanoparticles on the substrate surface. In the copper mode, the resolving power of the column was tested by injecting a mixtures from a library of prion-related peptides. The column was capable of separating prion peptides differing in number of octapeptide repeat units, presence of a hydrophilic tail, Q®A mutation or amide methylation. The IMAC material was also tested in the immobilized copper mode for resolving his-tagged protein aggregation variants and monoclonal antibody separation applications. In the immobilized iron mode, the column was test for the phosphopeptide enrichment applications. |
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General Papers
8:30 AM-11:10 AM, Sunday, 28 August 2005 Washington DC Convention Center -- 153, Oral
Sci-Mix
Division of Analytical Chemistry |