Monday, August 12, 2013

BioProcess International Poster Preview: Enhanced 2D Electrophoresis and Western Blotting Workflow for Reliable Evaluations of Anti-HCP Antibodies

Leading up to the BioProcess International Event, we'll be highlighting many of the posters you'll be able to find in the hall at the event taking place September 16-19, 2013 in Boston, MA. If you'd like to join us at the event, as a reader of this blog, when you register to join us and mention code BLOG13JP, you'll save 20% off the standard rate!  Would you like to submit a poster?  Do so here before August 19.

Poster: Enhanced 2D Electrophoresis and Western Blotting Workflow for Reliable Evaluations of Anti-HCP Antibodies

Presenter: Sricharan Bandhakavi, Bio-Rad Laboratories

About: Biologics and biosimilars can contain traces of proteins from the host cell line, which need to be minimized to prevent adverse immune reactions. The presence of host-cell proteins (HCP) is usually assayed by ELISA using a polyclonal antibody mixture raised against a population of proteins derived from the host cell background. This antibody should recognize as high a proportion as possible of the potential HCP’s in a given sample. A recommended method for evaluating the assay involves two-dimensional electrophoretic separation followed by Western blotting. Tools that simplify, standardize and accelerate 2-D electrophoresis and blotting have become available. Advanced electrophoresis and blotting instrumentation, stable and reproducible pre-cast gels, next-generation imaging and image analysis software all allow this assay to be performed in a reproducible manner in a relatively short time. We have undertaken to demonstrate this using a commercial anti-HCP antibody and samples derived from Chinese Hamster Ovary (CHO) cells. The 2 D electrophoresis procedure resulted in highly reproducible spot patterns and entire procedure was completed in less than two days. The software used (PDQuest) enabled the straightforward generation of percent coverage values for the antibody when used to probe both CHO cell lysate and CHO cell secreted protein.

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