On-Bead Antibody-Small Molecule Conjugation and pH Sensor Fluorescent Dye for Screening of Internalizing Antibodies for ADC Applications
On-Bead Antibody-Small Molecule Conjugation and pH Sensor Fluorescent Dye for Screening of Internalizing Antibodies for ADC Applications
30 Jun 2015This poster demonstrates a method that allows rapid screening of large number of antibodies for internalization in the cell. The method is a combination of two novel approaches: first, ‘on-bead’ antibody-small molecule conjugation that uses high capacity magnetic Protein A and Protein G beads to capture the antibody directly from the cell media and conjugate with a reactive small molecule, and second, conjugation of the antibody with a pH sensor dye that becomes fluorescent only when the antibody is internalized and trafficked to the endosomes and lysosomes.
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Magne™ Protein A and Magne™ Protein G Beads
Promega Corp.Magne™ Protein G and Magne™ Protein A Beads are magnetic affinity beads with high specificity and high capacity for purification of immunoglobulins from cell culture media, ascites and serum samples. These paramagnetic beads are composed of iron encapsulated in macroporous cellulose with low nonspecific binding. The magnetic beads use a novel attachment chemistry to immobilize recombinant Protein G or Protein A protein molecules in the same orientation on the surface of the bead. The oriented attachment is known to improve the functionality of immobilized proteins. These beads offer a convenient method for achieving high purity and high recovery of monoclonal and polyclonal antibodies from a variety of biological samples. The superb magnetic properties of Magne™ Protein G and Magne™ Protein A Beads allow rapid and efficient capture of antibodies either with manually processed samples or in a high-throughput manner using the Promega ReliaPrep™ LV 32 HSM Instrument or a robotic platform such as the Beckman Coulter Biomek® FX. Magne™ Protein A and Magne™ Protein G Beads: High purity and rapid, efficient antibody recovery Low nonspecific binding Ease of handling and cost effectiveness High binding capacity1 (≥25mg) of the beads for Protein A and Protein G Optimized performance