Standard Operating Procedure for Preparation of Lyophilized Enzyme Products
1) Purpose
This SOP outlines the procedure for preparing lyophilized enzyme products to maintain enzyme stability and bioactivity during freeze-drying and subsequent storage.
2) Scope
This SOP applies to all personnel involved in the formulation, preparation, and testing of enzyme products that undergo lyophilization, including formulation scientists, operators, and QA personnel.
3) Responsibilities
- Formulation Scientists: Responsible for selecting stabilizers and buffers to maintain enzyme stability and activity during lyophilization.
- Operators: Responsible for preparing the enzyme solution as per the Batch Manufacturing Record (BMR).
- QA Personnel: Responsible for validating enzyme activity post-lyophilization and ensuring proper documentation.
4) Procedure
4.1 Enzyme Formulation
4.1.1 Criteria for Enzyme Stability
- 4.1.1.1 The formulation must protect the enzyme from denaturation and loss of activity during freeze-drying. Common stabilizers such as sugars (e.g., sucrose, trehalose) or polyols (e.g., glycerol) may be used.
4.2 Preparation and Lyophilization
4.2.1 Solution Preparation
- 4.2.1.1 Weigh the enzyme and stabilizers accurately as per the BMR.
- 4.2.1.2 Dissolve the enzyme in a suitable buffer, ensuring that it is fully dissolved and that the stabilizers are evenly dispersed.
4.2.2 Sterile Filtration and Filling
- 4.2.2.1 Filter the solution through a sterile 0.22-micron filter to ensure sterility.
- 4.2.2.2 Aseptically fill sterile vials with the filtered solution, ensuring accurate fill volumes as per the BMR.
- 4.2.2.3 Freeze the vials at -80°C before proceeding with the lyophilization cycle.
4.3 Lyophilization and Post-Lyophilization Testing
- 4.3.1 Load the frozen vials into the lyophilizer and initiate the freeze-drying cycle as specified in the BMR.
- 4.3.2 After lyophilization, reconstitute the enzyme product and perform enzyme activity assays to ensure bioactivity is retained.
4.4 Documentation
Record all steps, including solution preparation, filtration, and post-lyophilization testing, in the Enzyme Product Lyophilization Logbook. Ensure that all records are verified by QA.
5) Abbreviations
- BMR: Batch Manufacturing Record
6) Documents
- Enzyme Product Lyophilization Logbook
7) References
- Pharmacopoeial guidelines for enzyme product formulation and testing
8) SOP Version
Version 1.0
Annexure
Enzyme Product Lyophilization Logbook Template
| Date | Enzyme Name | Batch No. | Stabilizer Used | Enzyme Activity Test | Operator Initials | QA Verification |
|---|---|---|---|---|---|---|
| DD/MM/YYYY | Enzyme Name | Batch Number | Stabilizer Name | Pass/Fail | Operator Name | QA Initials |