Application of trypsin digested-polypeptone medium in preparation of diphtheria vaccine adsorbed
10.13200/j.cnki.cjb.004536
- VernacularTitle:聚蛋白胨胰酶消化液培养基在吸附白喉疫苗制备过程中的应用
- Author:
WEN Jiana
- Publication Type:Journal Article
- Keywords:
Trypsin digested-polypeptone medium;
Diphtheria vaccine adsorbed;
Modified Linggood's medium;
Diphtherotoxin(DT);
Diphtheria toxiod
- From:
Chinese Journal of Biologicals
2025;38(08):902-907
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the possibility of using trypsin digested-polypeptone medium to prepare diphtheria vaccine adsorbed, so as to improve the quality and safety of the vaccine.Methods Corynebacterium diphtheriae(C.diphtheriae) was cultured in trypsin digested-polypeptone medium(polypeptone group) and modified Linggood's medium(modified Linggood's group), and three batches were cultured. The culture solution of C.diphtheriae was collected, and the toxin production detection, SDS-PAGE analysis and viable bacteria count were performed. Diphtherotoxin(DT) was extracted, and the 50% tissue culture infective dose(TCID50) was measured, and the logarithm was taken. After formaldehyde detoxification treatment, DT was tested for purity and analyzed by SDS-PAGE. Diphtheria vaccine adsorbed was prepared according to the corresponding method of Chinese Pharmacopoeia(VolumeⅢ, 2020 edition), and the vaccine titer was detected.Results The mean toxin production of three batches of culture solution of C.diphtheriae in polypeptone group was 177 Lf/m L, slightly lower than that of modified Linggood′s group(183 Lf/m L), and there was no statistically significant difference between them(t = 2. 00, P > 0. 05).The relative molecular weights of toxin protein and diphtheria toxoid obtained from two kinds of culture media were about 60 000. The mean values of A600and viable bacteria, lg TCID50of DT, purity of diphtheria toxoid and titer of diphtheria vaccine adsorbed in modified Linggood's group were significantly higher than those in polypeptone group(t = 9. 390, 31. 180,4. 914, 44. 540 and 4. 510, respectively, each P < 0. 05), while all the indicators in polypeptone group met the relevant requirements of Chinese Pharmacopoeia(VolumeⅢ, 2020 edition). Conclusion Trypsin digested-polypeptone medium is expected to replace the traditional modified Linggood's medium for the preparation of diphtheria vaccine adsorbed, while the preparation technology needs to be further optimized to improve the vaccine quality.
