Lot-to-Lot Quality Control Monitoring of Monoclonal Antibodies

As described in the FDA/CBER “Points to Consider” (PTC), each lot of product should be Quality-Control monitored and tested (21 CFR 600.3(x)) at specific intermediate production steps, as follows:

Unprocessed Bulk Lots

The following tests should be performed on each crude material harvest:

  • Sterility
  • HPLC-SEC
  • Adventitious Virus Contamination (with three indicator cell line
  • Mouse Antibody Production (MAP) or Hamster Antibody Production (HAP)
  • Retroviral Contamination
Purified Product Lots

Routine testing of purified bulk lots of antibody should include the following:

  • Purity (SDS-PAGE reduced and non-reduced)
  • HPLC-SEC
  • Isoelectric Focusing (IEF)
  • Sterility
  • Viral and Retroviral Contamination (if found in cell banks)
  • DNA Content (must be less than 100pg/dose)
  • Western Blot (recommended) - pH and Appearance (recommended)
Final-Filled Product Lots*

The following tests should be performed on the contents of final containers from each filled lot (21 CFR 600.3(y)):

  • Protein Concentration
  • Polyacrylamide Electrophoresis (Native and Reduced)
  • Sterility
  • General safety test (21 CFR 600.11)
  • HPLC-SEC (recommended)
  • Western Blot (recommended)
  • pH and Appearance (recommended)Isoenzyme

Complex formulations, freezing, and lyophilization may change the protein and may require additional testing.

Lot-to-Lot Quality Control Monitoring of Recombinant Proteins

As described in the FDA/CBER “Points to Consider” (PTC), each lot of product should be Quality-Control monitored and tested (21 CFR 600.3(x)) at specific intermediate production steps, as follows:

Unprocessed Bulk Lots

The following tests should be performed on each crude material harvest:

  • Sterility
  • HPLC-SEC
  • HPLC-Reversed Phase or Ion Exchange
  • Peptide Mapping
  • N-terminal Sequencing
  • Isoelectric Focusing (recommended)
Purified Product Lots

Routine testing of purified bulk lots of antibody should include the following:

  • Purity (SDS-PAGE reduced and non-reduced)
  • HPLC-SEC
  • Isoelectric Focusing (IEF)
  • Sterility
  • Amino Acid Analysis (recommended)
  • pH and Appearance (recommended)
Final-Filled Product Lots*

The following tests should be performed on the contents of final containers from each filled lot (21 CFR 600.3(y)):

  • Protein Concentration
  • Purity (SDS-PAGE reduced and non-reduced)
  • HPLC-SEC
  • Isoelectric Focusing (IEF)
  • Amino Acid Analysis (recommended)
  • Potency
  • Polyacrylamide Electrophoresis (Native and Reduced)
  • HPLC-SEC (recommended)
  • Western Blot (recommended)
  • pH and Appearance (recommended) Isoenzyme

Complex formulations, freezing, and lyophilization may change the protein and may require additional testing.