Process development is an essential stage in vaccine development that cannot be overlooked. It serves as the fundamental basis for the subsequent production, which encompasses crucial steps such as cell culture, fermentation, and various purification techniques. During the cell culture stage, the desired vaccine product is retained within the cells and does not naturally exit. Therefore, specific measures must be implemented to disrupt the cells and liberate the product into the surrounding liquid for subsequent purification procedures, which is called cell lysis.
Chemical and enzymatic lysis methods, such as SDS and lysozyme treatment, are usually gentler. Mechanical lysis, on the other hand, is a common method that produces cells of better uniformity through repeated freeze-thaw cycles. Compared to chemical lysis, the repeated freeze-thaw method is more efficient. To implement this method, cells need to be frozen below -20°C, thawed at room temperature, and this process is repeated several times. The principle behind this method is that ice formation within the cells and increased salt concentration in the remaining cell fluid can cause swelling and lead to cell structure fragmentation. In fact, the secret to successful lysis lies in the cryopreservation capacity of containers.
Although the freeze-thaw method is considered one of the most ideal methods for lysis, it poses a major challenge to the container as it must be resistant to frost. Due to the principle of thermal expansion and contraction, slight deformation, which is a normal and unavoidable phenomenon, may occur in the bottom or top layer of cell culture vessels after being sealed with a sealing cap and frozen.
Here is where NEST 10-layer BioFactory, -20 °C Frost Resistance jumps in, an ideal cell culture vessel for cryo-preservation at the temperature of -18°C to -20 °C without leakage, which has undergo strict validations and performance testings.
Freeze resistance performance ★★★★★.
NEST 10-layer BioFactory, -20 °C Frost Resistance, boasting a remarkable -20°C frost resistance, utilizes the revolutionary adhesive bonding technique, allowing it to effortlessly endure freezing temperatures ranging from -18°C to -20°C. This cutting-edge feature not only enhances its ability to withstand extreme cold but also significantly improves its leak-proof capacity.
Protocals
1.Take three NEST 10-layer BioFactories, add crystal violet solution at 200mL per layer after irradiation, then tighten the sealing cap.
2.Place in a freezer of -18°C to -20 °C for 48 hours.
3.After complete thawing, lay flat for 12 hours, and place on the side (each side for 2 hours) to observe for any leakage. After irradiation, pressurize it at 8Kpa for 90 seconds, repeat twice with an interval of at least 4 hours. If there is no air leakage, it is considered a qualified product.
First cycle of freeze-thaw
Second cycle offreeze-thaw
Third cycle offreeze-thaw
Summary
NEST 10-layer BioFactory, -20 °C Frost Resistance, passed the sealing test without any leakage. After three cycles of freeze-thaw at -18°C to -20 °C, slight deformation occurred in the bottom or top layer of the product, but there was no leakage, which is a testament to its excellent freeze resistance performance.
After irradiation, pressurize BioFactory at 8Kpa for 90 seconds, repeat twice with an interval of at least 4 hours. If there is no air leakage, it is considered a qualified product.
NEST 10-layer BioFactory, -20 °C Frost Resistance, undergoes a full set of inspection for sealing performance during production to ensure that every BioFactory has good sealing performance.
