Vaccine pretreatment for quantification of 146S antigen in foot-and-mouth disease vaccines by high performance size exclusion chromatography.

Yanmin Song; Yanli Yang; Zhiguo SU; Lili Liu; Yuanyuan Zhu; Yuan XU; Xingqi Zou; Qizu Zhao; Songping Zhang

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Vaccine pretreatment for quantification of 146S antigen in foot-and-mouth disease vaccines by high performance size exclusion chromatography.

Author: Yanmin SONG ¹ ; Yanli YANG ¹ ; Zhiguo SU ¹ ; Lili LIU ¹ ; Yuanyuan ZHU ² ; Yuan XU ² ; Xingqi ZOU ² ; Qizu ZHAO ² ; Songping ZHANG ¹
Author Information

1. State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China.
2. China Institute of Veterinary Drug Control, Beijing 100081, China.
Publication Type:Journal Article
Keywords: 146S; enzymatic digestion; foot-and-mouth disease vaccine; quantification; size exclusion chromatography
MeSH: Animals; Chromatography, Gel; Foot-and-Mouth Disease; Foot-and-Mouth Disease Virus; Reproducibility of Results; Viral Vaccines
From: Chinese Journal of Biotechnology 2019;35(8):1441-1452
CountryChina
Language:Chinese
Abstract: We developed a pre-treatment method to remove interfering substances during quantification of 146S antigens in foot-and-mouth disease (FMD) vaccines by high performance size exclusion chromatography (HPSEC). Three methods, including ultracentrifugation, PEG precipitation and nuclease digestion, were optimized and compared for removal efficiency of the interfering impurities in FMD vaccines. Under optimized conditions, the 146S contents in two batches of FMD vaccines were determined to be 7.1 and 7.6 μg/mL by ultracentrifugation, 9.7 and 10.4 μg/mL by PEG precipitation, and 10.5 and 10.4 μg/mL by nuclease digestion. The optimal condition for nuclease digestion using Benzonase determined by response surface method was as follows: appending Benzonase into 200 μL of antigen phase to a final concentration of 421 U/mL and incubating at 25.1 °C for 1.29 h. This method has advantages including efficient removal of the interfering impurities, fast processing speed, and mild operating conditions. Then 12 bathes of FMD vaccines with different serotypes produced by 4 manufacturers were tested to verify the established treatment method. Results showed the method was applicable to various FMD vaccines with good reproducibility (RSD<5.3%, n=3). The developed method removed interference from impurities during quantification of 146S, and therefore would broaden the application of HPSEC in vaccine quality control and ensure the accuracy and reliability.