Validation of Pretreatment Methods for the In-Process Quantification of Foot-and-Mouth Disease Vaccine Antigens

Foot-and-mouth disease (FMD), caused by the FMD virus (FMDV), is controlled by vaccine policy in many countries. For vaccine potency, the content of intact virus particles (146S antigens) is critical, and the sucrose density gradient (SDG) fractionation is the gold standard for the quantification of...

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Autores principales:	Ah-Young Kim, Sun Young Park, Sang Hyun Park, Jong Sook Jin, Eun-Sol Kim, Jae Young Kim, Jong-Hyeon Park, Young-Joon Ko
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	foot-and-mouth disease (FMD) pretreatment quantification vaccine antigen SE-HPLC Medicine R
Acceso en línea:	https://doaj.org/article/56bdcd52039a4b2ea7bd5dcf122b54f5
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id	oai:doaj.org-article:56bdcd52039a4b2ea7bd5dcf122b54f5
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spelling	oai:doaj.org-article:56bdcd52039a4b2ea7bd5dcf122b54f52021-11-25T19:11:38ZValidation of Pretreatment Methods for the In-Process Quantification of Foot-and-Mouth Disease Vaccine Antigens10.3390/vaccines91113612076-393Xhttps://doaj.org/article/56bdcd52039a4b2ea7bd5dcf122b54f52021-11-01T00:00:00Zhttps://www.mdpi.com/2076-393X/9/11/1361https://doaj.org/toc/2076-393XFoot-and-mouth disease (FMD), caused by the FMD virus (FMDV), is controlled by vaccine policy in many countries. For vaccine potency, the content of intact virus particles (146S antigens) is critical, and the sucrose density gradient (SDG) fractionation is the gold standard for the quantification of 146S antigens. However, this method has several drawbacks. Although size-exclusion high-performance liquid chromatography (SE-HPLC) was introduced to replace the classic method, its application is generally confined to purified samples owing to the interfering signals. Therefore, we aimed to develop optimal pretreatment methods for SE-HPLC quantification in less purified samples. Crude virus infection supernatant (CVIS) and semi-purified samples with PEG precipitation (PEG-P) were used. Chloroform pretreatment was essential to remove a high level of non-specific signals in CVIS, whereas it caused loss of 146S antigens without the distinctive removal of non-specific signals in PEG-P. Benzonase pretreatment was required to improve the resolution of the target peak in the chromatogram for both CVIS and PEG-P. Through spiking tests with pure 146S antigens, it was verified that the combined pretreatment with chloroform and benzonase was optimal for the CVIS, while the sole pretreatment of benzonase was beneficial for PEG-P.Ah-Young KimSun Young ParkSang Hyun ParkJong Sook JinEun-Sol KimJae Young KimJong-Hyeon ParkYoung-Joon KoMDPI AGarticlefoot-and-mouth disease (FMD)pretreatmentquantificationvaccine antigenSE-HPLCMedicineRENVaccines, Vol 9, Iss 1361, p 1361 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	foot-and-mouth disease (FMD) pretreatment quantification vaccine antigen SE-HPLC Medicine R
spellingShingle	foot-and-mouth disease (FMD) pretreatment quantification vaccine antigen SE-HPLC Medicine R Ah-Young Kim Sun Young Park Sang Hyun Park Jong Sook Jin Eun-Sol Kim Jae Young Kim Jong-Hyeon Park Young-Joon Ko Validation of Pretreatment Methods for the In-Process Quantification of Foot-and-Mouth Disease Vaccine Antigens
description	Foot-and-mouth disease (FMD), caused by the FMD virus (FMDV), is controlled by vaccine policy in many countries. For vaccine potency, the content of intact virus particles (146S antigens) is critical, and the sucrose density gradient (SDG) fractionation is the gold standard for the quantification of 146S antigens. However, this method has several drawbacks. Although size-exclusion high-performance liquid chromatography (SE-HPLC) was introduced to replace the classic method, its application is generally confined to purified samples owing to the interfering signals. Therefore, we aimed to develop optimal pretreatment methods for SE-HPLC quantification in less purified samples. Crude virus infection supernatant (CVIS) and semi-purified samples with PEG precipitation (PEG-P) were used. Chloroform pretreatment was essential to remove a high level of non-specific signals in CVIS, whereas it caused loss of 146S antigens without the distinctive removal of non-specific signals in PEG-P. Benzonase pretreatment was required to improve the resolution of the target peak in the chromatogram for both CVIS and PEG-P. Through spiking tests with pure 146S antigens, it was verified that the combined pretreatment with chloroform and benzonase was optimal for the CVIS, while the sole pretreatment of benzonase was beneficial for PEG-P.
format	article
author	Ah-Young Kim Sun Young Park Sang Hyun Park Jong Sook Jin Eun-Sol Kim Jae Young Kim Jong-Hyeon Park Young-Joon Ko
author_facet	Ah-Young Kim Sun Young Park Sang Hyun Park Jong Sook Jin Eun-Sol Kim Jae Young Kim Jong-Hyeon Park Young-Joon Ko
author_sort	Ah-Young Kim
title	Validation of Pretreatment Methods for the In-Process Quantification of Foot-and-Mouth Disease Vaccine Antigens
title_short	Validation of Pretreatment Methods for the In-Process Quantification of Foot-and-Mouth Disease Vaccine Antigens
title_full	Validation of Pretreatment Methods for the In-Process Quantification of Foot-and-Mouth Disease Vaccine Antigens
title_fullStr	Validation of Pretreatment Methods for the In-Process Quantification of Foot-and-Mouth Disease Vaccine Antigens
title_full_unstemmed	Validation of Pretreatment Methods for the In-Process Quantification of Foot-and-Mouth Disease Vaccine Antigens
title_sort	validation of pretreatment methods for the in-process quantification of foot-and-mouth disease vaccine antigens
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/56bdcd52039a4b2ea7bd5dcf122b54f5
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Validation of Pretreatment Methods for the In-Process Quantification of Foot-and-Mouth Disease Vaccine Antigens

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