Reliable, Streamlined 2-D Western Blot Workflow for Evaluation of Antibodies Developed for Detection of Host Cell Proteins

July 29, 2013 at 2:40 pm Leave a comment

The near complete removal of host cell protein (HCP) impurities from biologics (therapeutic protein drugs) is essential for patient safety and treatment efficacy. Hence, biologics are routinely monitored for levels of residual HCPs by a highly sensitive ELISA/immunoassay with a polyclonal antibody reagent raised against a wide range of potential HCP impurities (anti-HCP antibodies). To ensure accuracy of the ELISA, the range of HCP impurities detectable by the anti-HCP antibodies must be evaluated. Although 2-D electrophoresis (2-DE) and western blotting (described in detail in Bio-Rad’s Applications & Technologies section) is the gold standard method for evaluating anti-HCP antibodies, its adoption is hampered by its perceived lack of reproducibility and labor/time-intensive nature of the standard workflow. Here, we demonstrate an improved, optimized workflow for streamlined and highly reliable, 2-DE and western blotting-based evaluation of anti-HCP antibodies.

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Entry filed under: Bio-Rad Tech Note. Tags: , .

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