Stability of EIA kits for determination of pertussis antigens and their application in the production process of pertussis vaccine

Abstract

Objective To evaluate the stability of 3 kinds of EIA kits for detecting pertussis toxin (PT),filamentous haemagglutinin (FHA) and pertactin (Prn) and their application in the quality control of acellular pertussis vaccine (aPV) production.Methods The stability of EIA kits were evaluated by heat accelerated test (37 ℃ for 3 days) and long-term stability test (4 ℃ for 6 months),using the correlation coefficient of standard curve,the P/N value and the recovery of the quality control material.The contents of PT,FHA and Prn in the fermentation,purification and adsorption stages during the production process of aPV were detected by these 3 kinds of EIA kits,and the feasibility of using them in the quality control of aPV production were determined.Results Heat accelerated stability test and long-term stability test showed that quality parameters of 3 kinds of EIA kits all met the requirement.The detection results of pertussis antigens in the different production stages of pertussis vaccine obtained by 3 kinds of EIA kits showed that the fermentation end of Bordetella pertussis was at 42-46 h,and the purification recovery rates were 49.24％,56.12％ and 63.65％ for 3 lots of PT antigen,respectively,68.75％,55.60％ and 49.76％ for 3 lots of FHA antigen,respectively,and 75.73％,60.63％ and 50.10％ for 3 lots of Prn antigen,respectively.The antigen absorption rates of aluminum adjuvant with separate absorption were higher than those with mixed adsorption,but there was no difference in potency between both vaccines prepared by separate and mixed adsorption.Conclusion Three kinds of EIA kits for detecting PT,FHA and Prn have good stability and can be used for the quality control of aPV production. Key words: Pertussis vaccine; Immunoenzyme techniques; Quality control