Why use 2D Wb for Host Cell Protein (HCP) antibody coverage detection instead of ELISA, LCMSMS, or AAE?
Host Cell Protein (HCP) detection through 2D Western blot typically employs fluorescent labeling, allowing for the generation of HCP and WB images from the same gel and membrane. Data reports provide HCP, Western blots, and HCP/Western overlay images, directly displaying comprehensive antibody coverage that ELISA or LCMSMS cannot offer. Furthermore, our large-scale 2D provides high-resolution images of HCP and its modified forms that ELISA or LCMSMS cannot achieve, significantly enhancing coverage sensitivity and accuracy. AAE is based on the immune binding of antibodies to antigens under native conditions and has limitations in quantitative antibody coverage: False positives: a) Proteins directly or indirectly related to antibody-binding proteins; b) Antigens present in HCP antibodies; c) Co-purified serum proteins in HCP antibodies. False negatives: a) Protein complexes may block antibody binding; b) Some bound HCPs or their degradation fragments cannot be eluted. Additionally, AAE requires combination with downstream LCMSMS or 2D Western blot, and such an extended process introduces more variations and inconsistencies.
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