Lysyl endopeptidase lyophilization and storage protection agent
A technology of lysyl endopeptidase and endonuclease, which is applied in the field of medicine and can solve problems such as the mixture and method of lysyl endopeptidase that have not been disclosed
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Embodiment 1
[0060] Sucrose, mannitol, glycine or their salt protective agents were prepared at a concentration of 20%, and stored at -20°C for future use. This embodiment adopts 3 kinds of buffer concentrations, respectively 5 / 20 / 50mmol / L PBS buffer and Tris-HCl buffer, each concentration contains 7 pH values, pH4 / 5 / 6 / 7 / 8 / 9 / 10. When the pH value was 4 / 5 / 6 / 7, it was adjusted with PBS buffer solution, and when the pH value was 8 / 9 / 10, it was adjusted with Tris-HCl buffer solution. Take 84x3 vials, and add 100 μl of corresponding protective agent, 100 μl of lysyl endopeptidase solution, and 1800 μl of corresponding buffer solution to each vial according to the table. Add 100 μl of lysyl endopeptidase solution, 1900 μl of the corresponding buffer, then shake well and lyophilize. After freeze-drying, take it out and dissolve it with 2ml of ultrapure water, carry out the reaction according to the above-mentioned enzyme digestion reaction system, and then analyze the result by liquid phase. ...
Embodiment 2
[0069] On the basis of Example 1, several protective agents such as dextran, lactose, polyethylene glycol 4000, lysine or its salts were added. Each protective agent was still prepared at a concentration of 20%, and stored at -20°C for future use. The method of adding the composite protective agent is as in the above table. According to the results of Example 1, the buffer system is preferably pH5, 50mmol / LPBS and pH10, 50mmol / LTris-HCl. Take 26x3 vials, and add 100 μl of corresponding protective agent, 100 μl of lysyl endopeptidase solution, and 1600 μl of corresponding buffer solution to each vial according to the table. Add 100 μl of lysyl endopeptidase solution and 1900 μl of corresponding buffer to the bottle, then shake well and lyophilize. After freeze-drying, take it out and dissolve it with 2ml of ultrapure water, carry out the reaction according to the above-mentioned enzyme digestion reaction system, and then analyze the result by liquid phase. When the results we...
Embodiment 3
[0077]On the basis of Example 2, trehalose, a protective agent, was added, prepared at a concentration of 20%, and stored at -20°C for future use. This experiment explores the effects of compound protectants with five concentrations of sucrose, dextran, trehalose and glycine or their salts on the enzyme activity. The protective agent was added according to the above table, and according to the results of Examples 1 and 2, the pH 10, 50 mmol / L Tris-Hcl buffer was used as the buffer in this experiment. Take 76x3 vials, add 100 μl of lysyl endopeptidase solution to each vial according to the table, and then number 1 from 0.2% sucrose, 0.2% glycine or their salts, 0.5% sucrose, 0.2% glycine or their salts Salt number 6, followed by 5% trehalose, 5% glycine or its salt number 75. 0.2% protectant needs to add 20 μL to the freeze-drying system, 0.5% protectant needs to add 50 μL to the freeze-drying system, 1% 100 μL of protective agent should be added to the freeze-drying system, 2...
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