Patent ID: 11911460
Assignee: SHANDONG WEIGAO LITONG BIOLOGICAL PRODUCTS CO., LTD.
Field: Pharmaceuticals (Chemistry)
Classification: CPC A  Y | IPC A

Claim 2:
3. A preparation method of the VLS vaccine according to claim 2, comprising the following steps:
(1) mixing the DHA-1, the DOTAP, the DOPC, and the mPEG-DTA-1-2K dissolved in absolute ethanol according to a molar ratio of (10-13):(27-29):(56-59):(1.6-1.9) to obtain a mixed solution; and
diluting the mRNA with 10 mM NaAc, and 0.0001% (w/v, trehalose (buffer D) to obtain a diluted mRNA;
(2) encapsulating the diluted mRNA with the mixed solution obtained in step (1) by a microfluidic device to produce an mRNA-LNP system;
(3) diluting the mRNA-LNP system obtained in step (2) with buffer D in a volume 30 times a volume of the mRNA-LNP system, and subjecting a resulting diluted system to an ultrafiltration (UF) concentration with a 100 kDa UF membrane to obtain a filtered mRNA-LNP system;
(4) using 500 mM acetic acid (HA) to adjust a pH of the filtered mRNA-LNP system to 6.2 to 6.8 to obtain an adjusted mRNA-LNP system;
(5) adding the S1 antigen protein expressed in the CHO cell and purified by chromatography to buffer D with a pH of 6.2 to 6.8 to prepare a protein preparation solution system; and
(6) mixing the adjusted mRNA-LNP system obtained in step (4) with the protein preparation solution system obtained in step (5), and subjecting a resulting mixed system to a rotary-mixing at room temperature for 30 min and then staying overnight at 4° C. to obtain a stock solution for the vaccine.