How to resolve coagulation of human fibrinogen

Fibrinogen coagulation can be treated by slow rewarming, physiological saline dilution, and professional instrument shaking dissolution. Fibrinogen coagulation is mainly related to factors such as low temperature, improper storage, and improper operation.

1. Slow rewarming:

Place the solidified fibrinogen in a constant temperature water bath at 25-37 ℃ and slowly heat it up to avoid direct high-temperature heating that may cause protein denaturation. During the reheating process, gently shake the container every 10 minutes to observe the dissolution state, which usually takes 1-2 hours for complete dissolution. This method is suitable for mild solidification caused by low temperature.

2. Dilution with physiological saline:

Slowly add 0.9% sterile physiological saline to the solidified sample in a 1:1 ratio, while gently stirring in the same direction with a glass rod. Dilution can reduce the viscosity of the solution and promote the redispersion of fibrinogen molecules. The operation should be carried out in a clean bench to avoid microbial contamination.

3. Instrument oscillation dissolution:

uses a professional vortex oscillator to oscillate at a low speed of 800-1000rpm, combined with 37 ℃ constant temperature conditions to assist dissolution. During the oscillation process, it is necessary to pause observation intermittently to avoid excessive mechanical force damaging the protein structure. This method is suitable for moderate solidification state and needs to be operated by laboratory professionals.

4. Filtration treatment:

For solidified substances that are difficult to completely dissolve, a 0.22 μ m sterile filter membrane can be used for negative pressure filtration. The filtrate needs to undergo electrophoresis detection to confirm the integrity of fibrinogen molecules. This method may result in the loss of some active ingredients and is only used as an emergency response plan.

5. Preventive measures:

When storing, maintain a constant temperature of 2-8 ℃ and avoid light. Use a dedicated cold chain box for transportation. After opening, it should be used within 4 hours to avoid repeated freezing and thawing. Maintain room temperature balance for 30 minutes before operation, and avoid vigorous shaking to prevent the formation of bubbles during pipetting.

When using fibrinogen preparations in daily life, it is recommended to establish standardized operating procedures and equip with real-time temperature monitoring equipment. The dissolved solution needs to undergo coagulation activity testing. If the coagulation time exceeds the normal value by 20%, it should be discarded. For samples that require long-term preservation, it is recommended to pack them into small-sized cryovials, add stabilizers, and place them in a -80 ℃ deep low-temperature refrigerator, which can effectively maintain biological activity for 12 months. Regularly calibrate and maintain storage equipment to avoid protein denaturation caused by temperature fluctuations.