Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies.
Identifieur interne : 000604 ( PubMed/Checkpoint ); précédent : 000603; suivant : 000605Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies.
Auteurs : Chia Chiu Lim [Malaisie] ; Patrick C Y. Woo [République populaire de Chine] ; Theam Soon Lim [Malaisie]Source :
- Scientific reports [ 2045-2322 ] ; 2019.
Abstract
Antibody phage display has been pivotal in the quest to generate human monoclonal antibodies for biomedical and research applications. Target antigen preparation is a main bottleneck associated with the panning process. This includes production complexity, downstream purification, quality and yield. In many instances, purified antigens are preferred for panning but this may not be possible for certain difficult target antigens. Here, we describe an improved procedure of affinity selection against crude or non-purified antigen by saturation of non-binders with blocking agents to promote positive binder enrichment termed as Yin-Yang panning. A naïve human scFv library with kappa light chain repertoire with a library size of 109 was developed. The improved Yin-Yang biopanning process was able to enrich monoclonal antibodies specific to the MERS-CoV nucleoprotein. Three unique monoclonal antibodies were isolated in the process. The Yin-Yang biopanning method highlights the possibility of utilizing crude antigens for the isolation of monoclonal antibodies by phage display.
DOI: 10.1038/s41598-019-42628-6
PubMed: 30988390
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Woo</name><affiliation wicri:level="1"><nlm:affiliation>Department of Microbiology, The University of Hong Kong, Hong Kong SAR, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Microbiology, The University of Hong Kong, Hong Kong SAR</wicri:regionArea><wicri:noRegion>Hong Kong SAR</wicri:noRegion></affiliation></author><author><name sortKey="Lim, Theam Soon" sort="Lim, Theam Soon" uniqKey="Lim T" first="Theam Soon" last="Lim">Theam Soon Lim</name><affiliation wicri:level="1"><nlm:affiliation>Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800, Penang, Malaysia. theamsoon@usm.my.</nlm:affiliation><country xml:lang="fr">Malaisie</country><wicri:regionArea>Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800, Penang</wicri:regionArea><wicri:noRegion>Penang</wicri:noRegion></affiliation></author></analytic><series><title level="j">Scientific reports</title><idno type="eISSN">2045-2322</idno><imprint><date when="2019" type="published">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Antibody phage display has been pivotal in the quest to generate human monoclonal antibodies for biomedical and research applications. Target antigen preparation is a main bottleneck associated with the panning process. This includes production complexity, downstream purification, quality and yield. In many instances, purified antigens are preferred for panning but this may not be possible for certain difficult target antigens. Here, we describe an improved procedure of affinity selection against crude or non-purified antigen by saturation of non-binders with blocking agents to promote positive binder enrichment termed as Yin-Yang panning. A naïve human scFv library with kappa light chain repertoire with a library size of 10<sup>9</sup> was developed. The improved Yin-Yang biopanning process was able to enrich monoclonal antibodies specific to the MERS-CoV nucleoprotein. Three unique monoclonal antibodies were isolated in the process. The Yin-Yang biopanning method highlights the possibility of utilizing crude antigens for the isolation of monoclonal antibodies by phage display.</div></front></TEI><pubmed><MedlineCitation Status="In-Process" Owner="NLM"><PMID Version="1">30988390</PMID><DateRevised><Year>2020</Year><Month>04</Month><Day>14</Day></DateRevised><Article PubModel="Electronic"><Journal><ISSN IssnType="Electronic">2045-2322</ISSN><JournalIssue CitedMedium="Internet"><Volume>9</Volume><Issue>1</Issue><PubDate><Year>2019</Year><Month>04</Month><Day>15</Day></PubDate></JournalIssue><Title>Scientific reports</Title><ISOAbbreviation>Sci Rep</ISOAbbreviation></Journal><ArticleTitle>Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies.</ArticleTitle><Pagination><MedlinePgn>6088</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1038/s41598-019-42628-6</ELocationID><Abstract><AbstractText>Antibody phage display has been pivotal in the quest to generate human monoclonal antibodies for biomedical and research applications. Target antigen preparation is a main bottleneck associated with the panning process. This includes production complexity, downstream purification, quality and yield. In many instances, purified antigens are preferred for panning but this may not be possible for certain difficult target antigens. Here, we describe an improved procedure of affinity selection against crude or non-purified antigen by saturation of non-binders with blocking agents to promote positive binder enrichment termed as Yin-Yang panning. A naïve human scFv library with kappa light chain repertoire with a library size of 10<sup>9</sup> was developed. The improved Yin-Yang biopanning process was able to enrich monoclonal antibodies specific to the MERS-CoV nucleoprotein. Three unique monoclonal antibodies were isolated in the process. The Yin-Yang biopanning method highlights the possibility of utilizing crude antigens for the isolation of monoclonal antibodies by phage display.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Lim</LastName><ForeName>Chia Chiu</ForeName><Initials>CC</Initials><AffiliationInfo><Affiliation>Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800, Penang, Malaysia.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Woo</LastName><ForeName>Patrick C Y</ForeName><Initials>PCY</Initials><AffiliationInfo><Affiliation>Department of Microbiology, The University of Hong Kong, Hong Kong SAR, People's Republic of China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Lim</LastName><ForeName>Theam Soon</ForeName><Initials>TS</Initials><Identifier Source="ORCID">http://orcid.org/0000-0002-0656-3045</Identifier><AffiliationInfo><Affiliation>Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800, Penang, Malaysia. theamsoon@usm.my.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Analytical Biochemistry Research Centre, Universiti Sains Malaysia, 11800, Penang, Malaysia. theamsoon@usm.my.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2019</Year><Month>04</Month><Day>15</Day></ArticleDate></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Sci Rep</MedlineTA><NlmUniqueID>101563288</NlmUniqueID><ISSNLinking>2045-2322</ISSNLinking></MedlineJournalInfo><CitationSubset>IM</CitationSubset></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2018</Year><Month>07</Month><Day>16</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2019</Year><Month>04</Month><Day>04</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2019</Year><Month>4</Month><Day>17</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2019</Year><Month>4</Month><Day>17</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2019</Year><Month>4</Month><Day>17</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>epublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">30988390</ArticleId><ArticleId IdType="doi">10.1038/s41598-019-42628-6</ArticleId><ArticleId IdType="pii">10.1038/s41598-019-42628-6</ArticleId><ArticleId IdType="pmc">PMC6465254</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Malaisie</li><li>République populaire de Chine</li></country></list><tree><country name="Malaisie"><noRegion><name sortKey="Lim, Chia Chiu" sort="Lim, Chia Chiu" uniqKey="Lim C" first="Chia Chiu" last="Lim">Chia Chiu Lim</name></noRegion><name sortKey="Lim, Theam Soon" sort="Lim, Theam Soon" uniqKey="Lim T" first="Theam Soon" last="Lim">Theam Soon Lim</name></country><country name="République populaire de Chine"><noRegion><name sortKey="Woo, Patrick C Y" sort="Woo, Patrick C Y" uniqKey="Woo P" first="Patrick C Y" last="Woo">Patrick C Y. 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