Optimization for condition of adsorbing hepatitis A virus antigen with aluminum hydroxide adjuvant and evaluation of immunogenicity of adsorption products
- VernacularTitle:氢氧化铝佐剂与甲型肝炎病毒抗原吸附条件的优化及吸附产物免疫原性评价
- Author:
WANG Yibo
- Publication Type:Journal Article
- Keywords:
Hepatitis A virus(HAV);
Aluminum hydroxide;
Adjuvant;
Adsorption conditions;
Immunogenicity
- From:
Chinese Journal of Biologicals
2023;36(12):1425-1429
- CountryChina
- Language:Chinese
-
Abstract:
Objective To optimize the adsorption condition of hepatitis A virus(HAV)antigen by aluminum hydroxide[Al(OH)_3]adjuvant and evaluate the immunogenicity of the adsorbed products. Methods The final molar concentration of buffer(0. 01,0. 02 and 0. 04 mol/L),pH value(5. 5,6. 0,6. 5,7. 0,7. 5 and 8. 0),final molar concentration of sodium chloride(NaCl)(0. 075,0. 13,0. 15,0. 17,0. 45,0. 75 and 1. 0 mol/L),mixing mode(mixing after dilution and mixing directly without dilution)and dropping order[HAV stock solution into Al(OH)_3 solution,and Al(OH)_3 solution into HAV stock solution]were selected for single factor screening. Taking Al(OH)_3 adsorption rate as the evaluation index,the selected factors were optimized by orthogonal test. NIH mice were immunized intraperitoneally with adsorbed products prepared under optimal and suboptimal combination conditions with 0. 5 mL/mouse,10 mice in each group with half male and half female. After 28 d of immunization,the blood samples were collected from eyeball or heart,and the serum was separated. The HAV antibody level was detected by ELISA,and the immunogenicity of adsorbed products was evaluated by the seroconversion rate of mouse serum antibody. Results The results of single factor screening test showed that the final molar concentration of buffer,pH value and final molar concentration of NaCl had significant influence on the adsorption effect(F = 6. 582,14. 007 and 8. 612,respectively,each P < 0. 05),while the mixing mode and dropping order had no effect(t =-0. 696 and 1. 153,respectively,each P > 0. 05). The final molar concentration of buffer was 0. 01 and 0. 02 mol/L,the pH value was 5. 5 ～ 7. 0,and the final molar concentration of NaCl was 0. 13 ～ 0. 17 mol/L. The orthogonal test showed that the order of the influence on adsorption effect was the final molar concentration of buffer > pH value > NaCl final molar concentration. The optimal combination was pH 6. 3 + final molar concentration of buffer 0. 01 mol/L + NaCl molar final concentration 0. 15 mol/L,and the suboptimal combination was pH 5. 8 + final molar concentration of buffer 0. 02 mol/L +NaCl final molar concentration 0. 17 mol/L. The seroconversion rate of serum antibody in mice immunized with the adsorbed products prepared by the optimal and suboptimal combination was both 100%. Conclusion The adsorbed products prepared under the optimized conditions have good immunogenicity,which lays a foundation of the research of inactivated hepatitis A vaccine
