2-D DIGE Western Blot for HCP Detection and Quantification Launched

BioGenes GmbH, a leader in host cell protein (HCP) assay development for biomanufacturing processes, announce the availability of a 2-D DIGE Western Blot service for highly accurate determination and electronic quantification of HCP coverage.

2-D DIGE Western Blot is a DIGE based fluorescent two-dimensional gel electrophoresis combined with a Western Blot that detects whole protein and HCP antibody signals on the same membrane. It allows the protein spots to be aligned perfectly with immuno-detected spots, thus eliminating any gel-to-membrane variations. 2-D DIGE Western Blot uses two CyDye fluorescent dyes for labelling of total protein and HCP antibodies which enable signal detection at different wavelengths. CyDye fluorescence labelling is very sensitive and produces sharp and clear spots allowing an accurate and fully electronic analysis.

HCPs comprise the majority of protein contaminants derived from the manufacturing of therapeutic proteins and vaccines. HCPs are often immunogenic, can alter the therapeutic efficacy of a therapeutic drug substance and may affect patient safety. Therefore, the detection and quantification of HCPs is a major precondition for clinical trials of therapeutic proteins in human beings.

ELISA is a widely used technique to measure the residual HCPs and assess product purity. The current technique for estimating coverage of the HCP antibodies to the total HCPs present in a cell line is the comparison of a Coomassie Blue stained 2-D gel electrophoresis with an immunologically stained 2-D Western Blot. However, this technique may produce errors in pattern alignment between gel and membrane, is less sensitive than fluorescence and the results cannot be evaluated electronically.