About: Development of Monoclonal Antibody and Diagnostic Test for Middle East Respiratory Syndrome Coronavirus Using Cell-Free Synthesized Nucleocapsid Antigen

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About: Development of Monoclonal Antibody and Diagnostic Test for Middle East Respiratory Syndrome Coronavirus Using Cell-Free Synthesized Nucleocapsid Antigen Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : covidontheweb.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Development of Monoclonal Antibody and Diagnostic Test for Middle East Respiratory Syndrome Coronavirus Using Cell-Free Synthesized Nucleocapsid Antigen
Creator	Ying, Tianlei Matsuyama, Shutoku Takeda, Makoto Fukushi, Shuetsu Kimura, Hirokazu Obuchi, Masatsugu Matsushima, Yuki Ryo, Akihide Chimuro, Tomoyuki Matsunaga, Satoko Yamaoka, Yutaro Kuroyama, Hiroyuki Adachi, Akio Kabayama, Kazuya
Source	Medline; PMC
abstract	Protein nativity is one of the most critical factors for the quality of antigens used as immunogens and the reactivities of the resultant antibodies. The preparation and purification of native viral antigens in conventional cell-based protein expression systems are often accompanied by technical hardships. These challenges are attributable mainly to protein aggregation and insolubility during expression and purification, as well as to very low expression levels associated with the toxicity of some viral proteins. Here, we describe a novel approach for the production of monoclonal antibodies (mAbs) against nucleocapsid protein (NP) of the Middle East respiratory syndrome coronavirus (MERS-CoV). Using a wheat germ cell-free protein synthesis system, we successfully prepared large amounts of MERS-CoV NP antigen in a state that was highly soluble and intact for immunization. Following mouse immunization and hybridoma generation, we selected seven hybridoma clones that produced mAbs with exclusive reactivity against MERS-CoV NP. Epitope mapping and subsequent bioinformatic analysis revealed that these mAbs recognized epitopes located within relatively highly conserved regions of the MERS-CoV amino-acid sequence. Consistently, the mAbs exhibited no obvious cross-reactivity with NPs derived from other related viruses, including SARS coronavirus. After determining the optimal combinations of these mAbs, we developed an enzyme-linked immunosorbent assay and a rapid immunochromatographic antigen detection test that can be reliably used for laboratory diagnosis of MERS-CoV. Thus, this study provides strong evidence that the wheat germ cell-free system is useful for the production of diagnostic mAbs against emerging pathogens.
has issue date	2016-04-20(xsd:dateTime)
bibo:doi	10.3389/fmicb.2016.00509
bibo:pmid	27148198
has license	cc-by
sha1sum (hex)	2b2b4954a960745a09009e9de6d1cb87358bfa6a
schema:url	https://doi.org/10.3389/fmicb.2016.00509
resource representing a document's title	Development of Monoclonal Antibody and Diagnostic Test for Middle East Respiratory Syndrome Coronavirus Using Cell-Free Synthesized Nucleocapsid Antigen
has PubMed Central identifier	PMC4837155
has PubMed identifier	27148198
schema:publication	Front Microbiol
resource representing a document's body	covid:2b2b4954a960745a09009e9de6d1cb87358bfa6a#body_text
is schema:about of	named entity 'PROTEIN AGGREGATION' named entity 'LABORATORY DIAGNOSIS' named entity 'EPITOPE MAPPING' named entity 'IMMUNOGENS' named entity 'APPROACH' named entity 'SYSTEM' named entity 'SYSTEMS' named entity 'ATTRIBUTABLE' named entity 'PRODUCTION' named entity 'PREPARATION' named entity 'CRITICAL' named entity 'NATIVE' named entity 'CHALLENGES' named entity 'STRONG' named entity 'WHEAT GERM' named entity 'PATHOGENS' named entity 'SELECTED' named entity 'SARS CORONAVIRUS' named entity 'VIRAL ANTIGENS' named entity 'SOLUBLE' named entity 'NPS' named entity 'SEVEN' named entity 'ANTIGEN' named entity 'CELL-FREE SYSTEM' named entity 'TEST' named entity 'IMMUNIZATION' named entity 'DERIVED' named entity 'CROSSREACTIVITY' named entity 'ONE OF' named entity 'SUBSEQUENT' named entity 'QUALITY' named entity 'RAPID' named entity 'VIRAL PROTEINS' named entity 'PROVIDES' named entity 'NPs' named entity 'recognized' named entity 'mAbs' named entity 'mAbs' named entity 'prepared' named entity 'reactivities' named entity 'antigens' named entity 'located' named entity 'Middle East Respiratory Syndrome Coronavirus' named entity 'MERS-CoV' named entity 'MONOCLONAL ANTIBODY' named entity 'CONVENTIONAL' named entity 'INTACT' named entity 'CELL' named entity 'REACTIVITY' named entity 'HYBRIDOMA' named entity 'ENZYME-LINKED IMMUNOSORBENT ASSAY' named entity 'OPTIMAL' named entity 'NUCLEOCAPSID PROTEIN' named entity 'MIDDLE EAST RESPIRATORY SYNDROME CORONAVIRUS' named entity 'GENERATION' named entity 'TOXICITY' named entity 'CLONES' named entity 'EXPRESSION' named entity 'COMBINATIONS' named entity 'CELL-FREE PROTEIN SYNTHESIS' named entity 'FOLLOWING' named entity 'PURIFICATION' named entity 'AMOUNTS' named entity 'PREPARED' named entity 'REGIONS' named entity 'DEVELOPMENT' named entity 'USING' named entity 'NUCLEOCAPSID'

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