Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies
Identifieur interne : 000620 ( Main/Exploration ); précédent : 000619; suivant : 000621Development 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
Url:
DOI: 10.1038/s41598-019-42628-6
PubMed: 30988390
PubMed Central: 6465254
Affiliations:
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Y." last="Woo">Patrick C. Y. Woo</name><affiliation wicri:level="1"><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="ISNI">0000000121742757</institution-id><institution-id institution-id-type="GRID">grid.194645.b</institution-id><institution>Department of Microbiology,</institution><institution>The University of Hong Kong,</institution></institution-wrap>Hong Kong SAR, People’s Republic of China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Hong Kong SAR</wicri:regionArea></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:aff id="Aff1"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 2294 3534</institution-id><institution-id institution-id-type="GRID">grid.11875.3a</institution-id><institution>Institute for Research in Molecular Medicine,</institution><institution>Universiti Sains Malaysia,</institution></institution-wrap>11800 Penang, Malaysia</nlm:aff><country xml:lang="fr">Malaisie</country><wicri:regionArea>11800 Penang</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="Aff3"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 2294 3534</institution-id><institution-id institution-id-type="GRID">grid.11875.3a</institution-id><institution>Analytical Biochemistry Research Centre,</institution><institution>Universiti Sains Malaysia,</institution></institution-wrap>11800 Penang, Malaysia</nlm:aff><country xml:lang="fr">Malaisie</country><wicri:regionArea>11800 Penang</wicri:regionArea></affiliation></author></analytic><series><title level="j">Scientific Reports</title><idno type="eISSN">2045-2322</idno><imprint><date when="2019">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p id="Par1">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 <italic>Yin-Yang</italic> panning. A naïve human scFv library with kappa light chain repertoire with a library size of 10<sup>9</sup> was developed. The improved <italic>Yin-Yang</italic> biopanning process was able to enrich monoclonal antibodies specific to the MERS-CoV nucleoprotein. Three unique monoclonal antibodies were isolated in the process. The <italic>Yin-Yang</italic> biopanning method highlights the possibility of utilizing crude antigens for the isolation of monoclonal antibodies by phage display.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Smith, G" uniqKey="Smith G">G Smith</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Frenzel, Andre" uniqKey="Frenzel A">André Frenzel</name></author><author><name sortKey="Kugler, Jonas" uniqKey="Kugler J">Jonas Kügler</name></author><author><name sortKey="Helmsing, Saskia" uniqKey="Helmsing S">Saskia Helmsing</name></author><author><name sortKey="Meier, Doris" uniqKey="Meier D">Doris Meier</name></author><author><name sortKey="Schirrmann, Thomas" uniqKey="Schirrmann T">Thomas Schirrmann</name></author><author><name sortKey="Hust, Michael" uniqKey="Hust M">Michael Hust</name></author><author><name sortKey="Dubel, Stefan" uniqKey="Dubel S">Stefan 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uniqKey="Christensen B">B Christensen</name></author><author><name sortKey="K Lvraa, S" uniqKey="K Lvraa S">S Kølvraa</name></author><author><name sortKey="Kristensen, P" uniqKey="Kristensen P">P Kristensen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="S Rensen, Md" uniqKey="S Rensen M">MD Sørensen</name></author><author><name sortKey="Kristensen, P" uniqKey="Kristensen P">P Kristensen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Even Desrumeaux, K" uniqKey="Even Desrumeaux K">K Even-Desrumeaux</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Pavoni, E" uniqKey="Pavoni E">E Pavoni</name></author><author><name sortKey="Vaccaro, P" uniqKey="Vaccaro P">P Vaccaro</name></author><author><name sortKey="Anastasi, Am" uniqKey="Anastasi A">AM Anastasi</name></author><author><name sortKey="Minenkova, O" uniqKey="Minenkova O">O Minenkova</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Jones, Ml" uniqKey="Jones M">ML Jones</name></author></analytic></biblStruct></listBibl></div1></back></TEI><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><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. Woo</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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