Preparation method of DNA hydrogel loaded with IL-33 as well as product and application of DNA hydrogel loaded with IL-33
A technology of IL-33 and hydrogel, applied in medical preparations with non-active ingredients, medical preparations containing active ingredients, pharmaceutical formulas, etc., to improve the therapeutic effect, promote wound healing, and reduce the cycle of drug application
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Embodiment 1
[0036] see figure 1 . The preparation method of a DNA hydrogel loaded with IL-33 described in this example includes the following steps:
[0037] S1) Preparation: including recombinant mouse IL-33 and 5 kinds of single-stranded DNA as shown in SEQ ID No.1, SEQ ID No.2, SEQ ID No.3, SEQ ID No.4 and SEQ ID No.5, The concentration of the recombinant mouse IL-33 is 0.5ng / μL;
[0038] S2) Dissolution: each of the 5 single-stranded DNAs described in S1) was dissolved in 1×PBS buffer, the final concentration of the obtained lysing solution was 1 mmol / L, and 1 mmol / L of MgCl was added to the 1×PBS buffer. 2 , the pH of the 1xPBS buffer is 7.2;
[0039] S3) Construction of the monomer: the solubilization obtained in S2) by dissolving the single-stranded DNA shown in SEQ ID No. 1, SEQ ID No. 2 and SEQ ID No. 3 in the 1xPBS buffer described in S2), respectively After the liquid is mixed, a mixture with a concentration of 250 μmol / L is first subjected to constant temperature treatment...
Embodiment 2
[0043] see figure 1 . The preparation method of a DNA hydrogel loaded with IL-33 described in this example includes the following steps:
[0044] S1) Preparation: including recombinant mouse IL-33 and 5 kinds of single-stranded DNA as shown in SEQ ID No.1, SEQ ID No.2, SEQ ID No.3, SEQ ID No.4 and SEQ ID No.5, The concentration of the recombinant mouse IL-33 is 1ng / μL;
[0045] S2) dissolving: the 5 kinds of single-stranded DNAs described in S1) are respectively dissolved in 1×PBS buffer solution, the final concentration of the obtained lysing solution is 2mmol / L, and 2mmol / L MgCl is added to this 1×PBS buffer solution 2 , the pH of the 1×PBS buffer is 8.0;
[0046] S3) Construction of the monomer: the solubilization obtained in S2) by dissolving the single-stranded DNA shown in SEQ ID No. 1, SEQ ID No. 2 and SEQ ID No. 3 in the 1xPBS buffer described in S2), respectively After the liquid is mixed, the mixture with a concentration of 300 μmol / L is first subjected to consta...
Embodiment 3
[0050] A non-denaturing polyacrylamide gel electrophoresis method described in this example is used to determine whether the Y monomer and L monomer described in S3) in Example 1 are successfully constructed, and the steps include:
[0051] a) Prepare PAGE gel electrophoresis buffer;
[0052] b) Preparation of separating gel and stacking gel required for non-denaturing polyacrylamide gel electrophoresis;
[0053] c) performing native polyacrylamide gel electrophoresis;
[0054] d) Analyze the electrophoresis results, and judge whether the Y monomer and the L monomer are successfully constructed according to the general standard analysis method.
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