Human fibrin in-vitro degradation method
A technology for fibrin and degradation products, applied in the field of in vitro degradation of human fibrin, can solve the problems of FDP and D-dimer purity and stability, complex methods, and inability to be used as quality control or standard products, etc.
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Embodiment 1
[0039] Embodiment 1 preparation process screening
[0040] 1. Raw material preparation and preparation
[0041] 1.1. Tris (Tris) sodium chloride degradation buffer: 50 mM Tris-HCl, 0.9% normal saline, 0.1% sodium benzoate, pH 8.5.
[0042] 1.2, 2× freeze-drying buffer: 4-hydroxyethylpiperazineethanesulfonic acid (Hepes) 4%, trehalose 1g / L, bovine albumin 70g / L, mannitol 40g / L, sodium benzoate 0.2wt%, pH 7.6-7.8;
[0043] 1 × lyophilization buffer: 4-hydroxyethylpiperazineethanesulfonic acid (Hepes) 2%, trehalose 0.5g / L, bovine albumin 35g / L, mannitol 20g / L, sodium benzoate 0.1wt%, pH 7.6-7.8;
[0044] 1.3, calcium chloride (CaCl 2 ) solution: 25mM CaCl 2 solution.
[0045] 1.4. Fibrinolytic enzyme: Fibrinolytic enzyme purchased from Sigma, reconstituted in pure water, frozen in aliquots and stored in a -20°C refrigerator.
[0046] 1.5. Thrombin: dissolved in normal saline to prepare 120U / ml. Add thrombin to a final concentration of 2-3U / mL each time.
[0047] 1.6, pla...
Embodiment 2
[0084] According to the result of embodiment 1, select following steps to degrade:
[0085] 1. Raw material preparation and preparation
[0086] 1.1 Tris (Tris) sodium chloride degradation buffer: 50 mM Tris-HCl, 0.9% normal saline, 0.1% sodium benzoate, pH 8.5.
[0087] 1.2 2× freeze-drying buffer: 4-hydroxyethylpiperazineethanesulfonic acid (Hepes) 4%, trehalose 1g / L, bovine albumin 70g / L, mannitol 40g / L, sodium benzoate 0.2wt%, pH 7.6-7.8;
[0088] 1 × lyophilization buffer: 4-hydroxyethylpiperazineethanesulfonic acid (Hepes) 2%, trehalose 0.5g / L, bovine albumin 35g / L, mannitol 20g / L, sodium benzoate 0.1wt%, pH 7.6-7.8;
[0089] 1.3 Calcium chloride (CaCl 2 ) solution: 25mM CaCl 2 solution.
[0090] 1.4 Plasmin: Plasmin purchased from Sigma, reconstituted in pure water, frozen in aliquots and stored in a -20°C refrigerator.
[0091] 1.5 Thrombin: dissolved in normal saline to make 120U / ml. Add thrombin to a final concentration of 2-3U / mL each time.
[0092] 1.6 Pla...
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