Development and verification of whole column imaging detection-capillary isoelectric focusing method for determination of isoelectric point of pertactin
Development and verification of whole column imaging detection-capillary isoelectric focusing method for determination of isoelectric point of pertactin
Objective To develop and verify a whole column imaging detection-capillary isoelectric focusing(WCID-CIEF)method for the determination of isoelectric point(pI)of pertactin(PRN).Methods The WCID-CIEF method for the determination of PRN antigen was developed by optimizing the parameters such as the focusing time and final concentration of samples in the WCID-CIEF process,and verified for the specificity,accuracy,repeatability,intermediate precision,durability and inter-batch consistency.Results The optimal focusing time of WCID-CIEF for the determination of PRN antigen pI was 1 min at 1 500 V and 3 min at 3 000 V.The optimal final concentration of PRN antigen was 300 pg/mL.The PRN antigen pI was about 6.035,and the blank matrix showed no interference peak in the position of each peak of antigen.The method had good specificity,accuracy,repeatability,intermediate precision,durability and consistency among batches.Conclusion The developed WCID-CIEF method is suitable for the pI detection and charge heterogeneity analysis of PRN antigen,which can provide basis for the characterization of PRN antigen and reference for the quality control in the process of development and production of related vaccines.
关键词
百日咳黏附素/等电点/全柱成像毛细管等电聚焦电泳/质量控制
Key words
Pertactin(PRN)/Isoelectric point(pI)/Whole column imaging detection-capillary isoelectric focusing(WCID-CIEF)/Quality control
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