A kind of activity stabilizer of recombinant human keratinocyte growth factor-2 mutant
An active stabilizer, keratinocyte technology, applied in the direction of fibroblast growth factor, growth factor/inducing factor, animal/human protein, etc., can solve problems such as poor maintenance of activity and infection
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Embodiment 1
[0028] The protective agents lactose, dextran 40 and glycine were dissolved in different buffer systems. After dissolution, the concentrations of lactose, dextran 40 and glycine were all 1wt%, 3wt% and 20mM respectively, and then rhKGF-2M was dissolved therein. After being placed in an environment of 2-8° C. for 2 weeks, its biological activity was measured, and the results are shown in Table 1. The combination of phosphate buffer and protective agent had the best stabilizing effect, and the rhKGF-2M after treatment still had 98% of the initial activity; the combined effect of citrate buffer and protective agent was second, but rhKGF-2M after treatment 2M still has more than 92% of the initial activity.
[0029] Table 1 The influence of the combination of different buffer systems and protective agents on the stabilizing effect
[0030]
Embodiment 2
[0032] First prepare 20mM phosphate buffer, the formula is as follows: disodium hydrogen phosphate: 15.4mM; sodium dihydrogen phosphate: 4.6mM; EDTA: 2mM; NaCl: 100mM; pH7.3. Protectants (lactose, dextran 40 and glycine) were then added to the buffer. And set different protective agent content combinations: dextran concentration is 2%, 3%, 4%; lactose concentration is 1%, 2%, 3%; glycine concentration is 10, 20, 50mM. Then add rhKGF-2M to different active stabilizers, so that the final concentration of rhKGF-2M is 3mg / ml, subpackage according to the specification of 10mg and freeze-dry under appropriate conditions, reconstitute after freeze-drying for activity Analyze and observe the active stabilizing effect of each formulation on rhKGF-2M, the results are shown in Table 2.
[0033] The results show:
[0034] (1) When the concentration of lactose in the stabilizer is 3wt%, it is difficult to control the moisture content of the freeze-dried product, so the concentration of lac...
Embodiment 3
[0040] Example 3: Stability study of rhKGF-2M
[0041] Prepare 20 mM phosphate buffer with a pH of 7.3 as follows: disodium hydrogen phosphate: 15.4 mM; sodium dihydrogen phosphate: 4.6 mM; EDTA: 2 mM; NaCl: 100 mM, dissolve lactose, dextran 40 and glycine in the buffer , make the final concentration of dextran be 3wt%, the concentration of lactose be 1wt%, the concentration of glycine be 20mM, obtain 1× active stabilizer, rhKGF-2M powder is added wherein, make the final concentration of rhKGF-2M be 3mg / ml, lyophilized.
[0042] Or, at first configure 20mMpH as 7.3 phosphate buffer according to the above method, then dissolve lactose, dextran 40 and glycine in the buffer, so that the concentration of lactose is 2wt%, the concentration of dextran 40 is 6wt%, and the concentration of glycine is 40 mM, resulting in a 2× active stabilizer. Dissolve rhKGF-2M in the above-mentioned phosphate buffer, and then mix it with 2× active stabilizer so that the final concentration of lacto...
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