Method of stabilizing protein C or activated protein C and stabilized composition obtained therefrom
A protein and activity technology, applied in the field of protein C or active protein C, can solve problems such as stability and reduced activity of active protein C not seen
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example 1
[0032] Effects of various counter ions on the stability of APC:
[0033] 2.5% of human serum albumin (hereafter also referred to simply as "HSA") was added to active protein C with an activity of 500 U / ml. The solution was then dialyzed against a solution consisting of 20 mM each of sodium citrate, sodium phosphate and sodium sulfate containing 0.7% NaCl and 0.067M glycine. After dialysis, each solution was kept at 37°C for 24 hours, after which the activity was measured. The results are listed in Table 1. Sodium citrate, sodium phosphate, sodium sulfate, the counterions tested showed similar satisfactory stability.
[0034] Table 1
[0035] Effects of various counter ions on the stability of APC
[0036] counter ion Activity retention ratio (%)
[0037] (37°C, 24 hours)
[0038] Sodium citrate 20mM 97.4
[0039] Sodium Phosphate 20mM 94.8
[0040] Sodium sulfate 20mM 92.2
example 2
[0042] Effects of amino acids on APC activity:
[0043] 2.5% HSA was added to the active protein C solution with an activity of 500 U / ml. The solution was then dialyzed using a buffer containing 0.7% sodium chloride and 0.05M sodium citrate each of glycine, alanine, lysine, arginine, aspartic acid and glutamic acid. After dialysis, each solution was kept at 37°C for 24 hours, and the activity was measured. The results are listed in Table 2. The above six amino acids all showed high stability, and did not destroy the stability of APC.
[0044] Table 2
[0045] Effects of various amino acids on the stability of APC
[0046] Amino Acid Activity Retention Ratio (%)
[0047] (0.5％) (37°C, 24 hours)
[0048] No amino acids 86.1
[0049] Glycine 97.7
[0050] Alanine 97.7
[0051] Lysine 107
[0052] Arginine 102
[0053] Aspartic Acid 108
[0054] Glutamate 106
example 3
[0056] Effects of joining an HSA:
[0057] Two solutions were prepared, all containing human active protein C (1700U / ml), 20mM citric acid, 0.7% sodium chloride and 0.067M glycine, one of which contained 2.5% HSA, and the other did not contain HSA. The solution was then placed at 37°C and 4°C. The activity at different times was determined by the method described herein, and the percentage of retained activity was obtained. The results are listed in Table 3.
[0058] table 3
[0059] Maintain activity percentage (%) under different conditions
[0060] At 37°C:
[0061] Placement time (hours)
[0062] 0 1 3 6 8 24
[0063] HSA(+) 100 - 99.2 - 101 97.4
[0064] HSA(-) 100 888 92.0 86.2 90.9 79.3
[0065] At 4°C:
[0066] Placement time (hours)
[0067] 0 1 3 5 7 14
[0068] HSA(+) 100 - 105 - - 98.1
[0069] HSA(-) 100 112 99.1 104 98.9 85.2
[0070] From the results shown in Table 3, it can be seen that th...
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