Production of monoclonal antibodies against AFLM (Ver-1), a middle key protein involved in aflatoxin biosynthesis

Aflatoxins are potent carcinogens, mutagens and teratogens, and are harmful to both humans and animals. As many as 30 genes are involved in aflatoxin biosynthesis. Among them, aflM (ver-1) gene was predicted to encode a 28-kDa NADPH-dependent ketoreductase (AFLM), which catalyzed middle enzymatic st...

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Autores principales:	Ting Wang, Qi Zhang
Formato:	article
Lenguaje:	EN
Publicado:	KeAi Communications Co., Ltd. 2021
Materias:	Aflatoxin biosynthesis AFLM Monoclonal antibodies Quantitative detection Genetics QH426-470
Acceso en línea:	https://doaj.org/article/1bf72e67a6394824a60f5640b602e3b6
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spelling	oai:doaj.org-article:1bf72e67a6394824a60f5640b602e3b62021-11-26T04:27:34ZProduction of monoclonal antibodies against AFLM (Ver-1), a middle key protein involved in aflatoxin biosynthesis2096-242810.1016/j.ocsci.2021.10.003https://doaj.org/article/1bf72e67a6394824a60f5640b602e3b62021-10-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2096242821000531https://doaj.org/toc/2096-2428Aflatoxins are potent carcinogens, mutagens and teratogens, and are harmful to both humans and animals. As many as 30 genes are involved in aflatoxin biosynthesis. Among them, aflM (ver-1) gene was predicted to encode a 28-kDa NADPH-dependent ketoreductase (AFLM), which catalyzed middle enzymatic steps in aflatoxin biosynthetic pathway. AFLM (Ver-1) was proved to be necessary for conversion of versicolorin A (VERA) to demethylsterigmatocystin (DMST) in aflatoxin B1 (AFB1) biosynthesis. For these reasons, aflM gene was cloned and specific monoclonal antibodies for AFLM was developed to better define potential pathways of AFLM involved in AFB1 biosynthesis. Monoclonal antibodies 11B2-1D7 and 3G5-4E7 were successfully screened out by immunizing mouse. Immunoblot analysis revealed that both had high sensitivity and specificity to identify native AFLM protein in A. flavus with detection limit of 11 ng/mL and 8 ng/mL respectively. These results showed that it was suitable for quantitative detection of AFLM in A. flavus isolate. Further investigation revealed that aflatoxin accumulations of various A. flavus were not dependent on AFLM biosynthesis. Overall, this is the first report for development for AFLM monoclonal antibody development and application in A. flavus quantitative detection.Ting WangQi ZhangKeAi Communications Co., Ltd.articleAflatoxin biosynthesisAFLMMonoclonal antibodiesQuantitative detectionGeneticsQH426-470ENOil Crop Science, Vol 6, Iss 4, Pp 201-205 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Aflatoxin biosynthesis AFLM Monoclonal antibodies Quantitative detection Genetics QH426-470
spellingShingle	Aflatoxin biosynthesis AFLM Monoclonal antibodies Quantitative detection Genetics QH426-470 Ting Wang Qi Zhang Production of monoclonal antibodies against AFLM (Ver-1), a middle key protein involved in aflatoxin biosynthesis
description	Aflatoxins are potent carcinogens, mutagens and teratogens, and are harmful to both humans and animals. As many as 30 genes are involved in aflatoxin biosynthesis. Among them, aflM (ver-1) gene was predicted to encode a 28-kDa NADPH-dependent ketoreductase (AFLM), which catalyzed middle enzymatic steps in aflatoxin biosynthetic pathway. AFLM (Ver-1) was proved to be necessary for conversion of versicolorin A (VERA) to demethylsterigmatocystin (DMST) in aflatoxin B1 (AFB1) biosynthesis. For these reasons, aflM gene was cloned and specific monoclonal antibodies for AFLM was developed to better define potential pathways of AFLM involved in AFB1 biosynthesis. Monoclonal antibodies 11B2-1D7 and 3G5-4E7 were successfully screened out by immunizing mouse. Immunoblot analysis revealed that both had high sensitivity and specificity to identify native AFLM protein in A. flavus with detection limit of 11 ng/mL and 8 ng/mL respectively. These results showed that it was suitable for quantitative detection of AFLM in A. flavus isolate. Further investigation revealed that aflatoxin accumulations of various A. flavus were not dependent on AFLM biosynthesis. Overall, this is the first report for development for AFLM monoclonal antibody development and application in A. flavus quantitative detection.
format	article
author	Ting Wang Qi Zhang
author_facet	Ting Wang Qi Zhang
author_sort	Ting Wang
title	Production of monoclonal antibodies against AFLM (Ver-1), a middle key protein involved in aflatoxin biosynthesis
title_short	Production of monoclonal antibodies against AFLM (Ver-1), a middle key protein involved in aflatoxin biosynthesis
title_full	Production of monoclonal antibodies against AFLM (Ver-1), a middle key protein involved in aflatoxin biosynthesis
title_fullStr	Production of monoclonal antibodies against AFLM (Ver-1), a middle key protein involved in aflatoxin biosynthesis
title_full_unstemmed	Production of monoclonal antibodies against AFLM (Ver-1), a middle key protein involved in aflatoxin biosynthesis
title_sort	production of monoclonal antibodies against aflm (ver-1), a middle key protein involved in aflatoxin biosynthesis
publisher	KeAi Communications Co., Ltd.
publishDate	2021
url	https://doaj.org/article/1bf72e67a6394824a60f5640b602e3b6
work_keys_str_mv	AT tingwang productionofmonoclonalantibodiesagainstaflmver1amiddlekeyproteininvolvedinaflatoxinbiosynthesis AT qizhang productionofmonoclonalantibodiesagainstaflmver1amiddlekeyproteininvolvedinaflatoxinbiosynthesis
_version_	1718409887645433856

Production of monoclonal antibodies against AFLM (Ver-1), a middle key protein involved in aflatoxin biosynthesis

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