Recommended Match Rate by 2-D Electrophoresis and Western Blotting

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Recommended Match Rate by 2-D Electrophoresis and Western Blotting


Question:

What is the recommended match rate (% of host cell proteins detectable by a sensitive protein stain and immunoreactive against the anti-HCP antiserum/antibodies) by 2-D electrophoresis and western blotting?


Answer:

Although there are no regulatory recommendations, the assumption is that the higher the match rate, the better the anti-HCP antiserum.

  • Some researchers require a match rate of ~80% before qualifying the antibodies for use in host cell protein ELISA
  • Lower match rates may be acceptable if the nondetectable impurities do not copurify with the final product or if they are monitored by orthogonal methods


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