Objective To optimize some key items in testing method for final product of freezed-dried live attenuated hepatitis Avaccine.
Methods Testing method for in fective titer:To establish micro-immunofluorescent method and compare its advantages with that of conventional cell culture ELISA method; Testing method for bovine serum protein residual content:To compare the reproducibility and accuracy between reverse indirecthemagglutination method and ELISA method; Testing method for water content:To establish "solvent-aid adding" method and the vaccine water content results were compared before and after solvent-aid adding.
Results (1) The sensitivity of the micro-immunofluorescent method and conventional cell culture ELISA method was in same level(P>0.05), but the former reproducibility was better.The results could be judged one week earlier; Leica Qwin fluorescence analysis software was helpful in getting standardization results.(2) The results of reverse indirecthemagglutination method were not stable, the variation coecficient value(CV)were 39.12% and 34.23% in two batches vaccine respectively(5 times testing).The accuracy and reproducibility of ELISA method we, reall better(CV 2.19%, 2.82%, reclamation 101.73%).(3)The CVin two batches vaccine results were decreased from 8132%, 9.03% to 1.85%, 1.82% respectively before and after the solvent-aid adding(5 times testing).
Conclusion Micro-immunofluorescent method may replace the routine cell culture ELISA method in infective titer testing; ELISA method is better than reverse indirecthemagglutination method for bovine serum protein residual content testing; The adding of solvent-aid can improve the reproducibility of water content testing.
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