The traditional 2D Western blot method has been a widely accepted method for HCP coverage determination in the past decades. However, regulatory agencies highly recommend the use of such orthogonal methods for comprehensive reagent characterization. Employing Immunoaffinity Chromatography (IAC) with subsequent 2D DIGE analysis, BioGenes offers a reliable method for coverage determination. In here, IAC serves as a superior method enabling HCP binding to an immobilized mixture of polyclonal anti-HCP antibodies. This enables antibody-antigen interaction at non-denaturing conditions, thereby overcoming the methodological limitation of the denaturing 2D Western blot coverage assessment. Following the chromatographic step, 2D DIGE is used to visualize and analyze the HCP spectrum of the elution fraction by direct comparison to the input sample of interest prior to IAC for HCP coverage determination (see also the Figure below for a schematic overview).
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If you want to learn more about BioGenes’ capacities for custom ELISA development, have a look here. Also, have a look at BioGenes’ webinar on custom HCP ELISA development for detailed insights in our strategy to determine the HCP impurities in your samples.