A1PI Purification Ethanol Modulation Lysine Truncation

The patent applies parameter changes by modulating the ethanol concentration during Cohn fractionation. By adjusting ethanol levels (e.g., using 20% ethanol instead of higher concentrations), the patent controls the activity of carboxypeptidases that cause C-terminal lysine truncation. This allows optimization of both A1PI concentration and the preservation of intact C-terminal lysine residues.

The patent uses carboxypeptidase inhibitors as intermediary substances to prevent unwanted C-terminal lysine truncation during the purification process. These inhibitors mediate between the purification steps and the protein structure, blocking the harmful enzymatic activity while allowing the beneficial concentration and purification effects to proceed.

The patent introduces carboxypeptidase inhibitors as intermediary substances during the purification process. These inhibitors mediate by blocking the enzymatic activity of carboxypeptidases, thereby preventing C-terminal lysine cleavage while allowing the purification process to continue. This maintains both ease of manufacture and the reliability of A1PI stability and efficacy.

The patent applies preliminary anti-action by adding carboxypeptidase inhibitors before the purification process begins or at early stages. This preventive measure counteracts the potential harmful effect of carboxypeptidase activity before it can cause significant C-terminal lysine truncation, thereby protecting A1PI integrity throughout the manufacturing process.

The patent applies parameter changes by optimizing the ethanol concentration parameter during Cohn fractionation. By using moderate ethanol concentrations (e.g., 20%) rather than high concentrations, the patent achieves sufficient A1PI concentration while minimizing the activation of carboxypeptidases that cause harmful lysine truncation. This represents an optimal balance between two competing parameters.

The patent converts the potential harm of carboxypeptidase activity into a benefit by using carboxypeptidase inhibitors. These inhibitors transform the problematic enzymatic activity into a controlled process, allowing the purification to proceed while preventing unwanted lysine truncation. The harmful effect is neutralized and the process is made beneficial.