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Development of High-Capacity Magnetic Beads for Antibody and Protein Purification

Part # PS209

Abstract

Rob Chumanov, Nidhi Nath, Becky Godat, Rod Flemming and Marjeta Urh
Promega Corporation, 2800 Woods Hollow Rd, Madison, WI 53711.

Recombinant proteins and antibodies are key components in therapeutics, diagnostics, clinical studies and biological research. These applications require access to large numbers of highly purified proteins and antibodies to screen for desirable properties. To address this challenge we developed a magnetic platform as a simple, robust and reproducible method to purify proteins and antibodies from a large panel of samples (1–96) of small to medium volume (50μl–50ml). A key element of the magnetic platform is the use of macroporous, cellulose-based magnetic beads offering high capacity, low non-specific binding and excellent magnetic response for manual or automated handling. We highlight the advantages of the magnetic platform using multiple examples. We present a high-throughput purification of antibodies from variety of samples including cell media, ascites fluid and serum using high-capacity magnetic Protein A and Protein G beads—a method that is readily automatable on robotic liquid handlers including Tecan and Beckman. We describe a novel on-bead antibody-small molecule conjugation method that combines antibody labeling and purification in a single workflow precluding the need for antibody pre-purification, concentration or dialysis prior to labeling. Lastly we present a high-throughput protein purification example for purifying multiple HIS-tagged proteins expressed in E. coli and purified with high capacity Magnetic HIS affinity beads.

Printed in USA.