Establishment and verification of purity detection method for receptor-binding domain dimer in SARS-CoV-2 recombinant protein vaccine
Objective To establish and verify a bioanalyzer(Agilent 2100)method for the determination of the receptor-binding domain(RBD)dimer purity in SARS-CoV-2 recombinant protein vaccine.Methods The protein in the SARS-CoV-2 recombinant vaccine was analyzed and then concentrated by ultrafiltration,and the purity of fully denatured and partially denatured(disulfide bond retention)proteins was detected by bioanalyzer(Agilent 2100).The established method was veri-fied for the applicability,specificity,reproducibility,precision and accuracy,and used to detect four batches of samples from manufacturer A.Results In the system applicability test,eight peaks were clearly distinguished from the baseline.The blank solvent had no interference at the peak position of the target component,indicating the good specificity.Under the fully dena-tured and partially denatured conditions,the RSDs of 12 repeated test results by two experimenters were both less than 1％,showing good repeatability and precision.The RSDs of the proportion of the main peak(1+2)and the main peak at three different protein concentrations(50％,80％and 100％)were 0.68％and 0.31％,respectively,and the accuracy was good.Among them,the results of the 10％protein concentration sample were unstable.The purity of RBD dimer in four batches was 70.5％,70.2％,73.2％and 69.6％,respectively.Conclusion The established method has good specificity,precision and accuracy,and can be used for the detection of protein purity in recombinant protein vaccines.
SARS-CoV-2Recombinant protein vaccineReceptor-binding domain(RBD)DimerProtein purity
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