Preparation and immunogenicity evaluation of a novel ACYW135 group meningococcal-cross-reacting material 197 conjugate vaccine
Objective To prepare a novel ACYW135 group meningococcal-cross-reacting material 197(CRM197)conjugate vaccine and evaluate its immunogenicity,so as to lay a foundation for the prevention and control of epidemic cerebrospinal meningitis.Methods Group A,C,Y and W135 meningococcal polysaccharide hydroxyl was activated into cyano group with 1-cyano-4-dimethylamino pyridinium tetrafluoroborate(CDAP),respectively.Adipic acid dihydrazide(ADH)was used as a linker to connect one end of the activated polysaccharide by cyanate ester to generate polysaccharide-ADH derivative with exposed hydrazide groups.The monovalent conjugate vaccines were obtained through amide bond coupling between 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl morpholinium chloride(DMTMM)and CRM197.Then the monovalent conjugate was mixed with different lyophilized excipients(sucrose solution,lactose solution,and sucrose+mannitol solution)in proportion respectively and lyophilized to prepare three ACYW135-CRM197 meningococcal conjugate vaccines.The male NIH mice were subcutaneously immunized with the vaccines at 0 and 14 d,with 0.2 mL/mouse,10 mice in each group.At 21 d after the initial immunization,the blood samples were collected from submandibular vein of the mice,and the serum was separated and detected for the specific antibody level by ELISA.Results The test results of A,C,Y and W135-CRM197 monovalent conjugate all met the requirements of the quality standards.ACYW135-CRM197 vaccine product formed obvious precipitation line with each group of polysaccharide antiserum.The average molecular mass was all in the range of 1 680 000-5 230 000.The seroconversion rates of IgG antibodies against A,C,Y,W135 polysaccharide of three ACYW135-CRM197 conjugate vaccines were all 100％.There was no significant difference in polysaccharide antibody level among ACYW135-CRM197(sucrose),ACYW135-CRM197(lactose)and ACYW135-CRM197(sucrose+mannitol)groups(t=0.249-1.157,eachP＞0.05).Conclusion In this study,a novel ACYW135-CRM197 conjugate vaccine was prepared and had good immunogenicity in mice.
Neisseria meningitidis(Nm)Cross-reacting material 197(CRM197)Conjugate vaccineImmunogenicity
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