Validating cleaning procedures protein
In these conditions, it is not possible to definitively conclude on the level of acceptable limit in residues on the surface of manufacturing equipment after cleaning.Reasonable justifications must be provided if different acceptance criteria are applied for equipment used for the upstream and downstream phases of the manufacturing process and establish the clearance of degradation impurities in the drug substance.Even if denaturation or degradation of biological active proteins lead generally to inactive fragments, the lack of identification of impurities does not guarantee the complete safety of the cleaning residues.In the Design Phase, laboratory studies may be conducted to assess hydrolysis capacity in cleaning conditions.Similarly, ion exchange chromatography will make it possible to eliminate degradation impurities generated by hydrolysis of the main product that are generally more polar and more soluble in water.As for affinity chromatography, without studies, it is not possible to clearly specify the level of clearance.But they don’t bring a clear position on acceptance criteria definition.
Because chemical stability of peptide bonds is very high, hydrolysis is not complete, the chemical treatment leads to amino acids, oligomers and fragments.
In order to identify major degradation residues, characterization studies should be performed.
Laboratory experiments in cleaning conditions (concentration of cleaning agent, temperature, contact time) make it possible to obtain the degradation residues which are characterized by LC-MS or by simpler methods such as SDS-PAGE.
The document reports that “typical values established a acceptance criteria by manufacturers are 1-2 ppm TOC for downstream process and 5-10 ppm TOC for upstream process” but in fact, no scientific rationale is established to justify these values.
As specified in GMP annex 15 for macromolecules and peptides, it is well known that biological products are sensitive to hydrolysis at extreme p H, ,  and/or high temperature.