A method for antigen dissociation of CA16 antigen in EV71-CA16 bivalent inactivated vaccine containing aluminium adjuvant
To develop a method for CA16 antigen dissociation and content determination of EV71-CA16 bivalent inactivated vaccine containing aluminiuma djuvant for the detection of CA16 antigen content.The ratio of sodium citrate,10％tramadol and 20％diethanolamine was fixed by single variable method to determine the best formula of dissociation solution,on this basis,the vaccine was treated with different incubation temperature and time,the content of CA16 antigen after dissociation was detected by ELISA,the antigen recovery rate was calculated,the best dissociation solution formula and dissociation con-ditions were determined,and its repeatability was investigated.The results showed that the best formula of dissociation solu-tion is 20％diethanolamine 1.25 mL,10％tralatone 50 μL,the concentration of sodium citrate is 14％,and the final volume is fixed to 10 mL with 0.01 M PBS,the best dissociation condition is incubation at room temperature(18-25℃)for 0.5 h.After dissociation,the antigen content of CA16 reached 1 800 U/mL,the recovery rate was more than 90％,the linear corre-lation coefficient R2 was more than 0.97,and the coefficient of variation of repeated tests was less than 5％.The results in-dicate that the dissociation method has high recovery,good repeatability and accurate detection results.
EV71-CA16 bivalent inactivated HFMD vaccineCoxsackievirus A 16(CA16)DissociationbufferAluminiumAdju-vantELISA
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