Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies
Identifieur interne : 000343 ( Pmc/Checkpoint ); précédent : 000342; suivant : 000344Development 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:
Links toward previous steps (curation, corpus...)
Links to Exploration step
PMC:6465254Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies</title><author><name sortKey="Lim, Chia Chiu" sort="Lim, Chia Chiu" uniqKey="Lim C" first="Chia Chiu" last="Lim">Chia Chiu 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></author><author><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><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></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">30988390</idno><idno type="pmc">6465254</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6465254</idno><idno type="RBID">PMC:6465254</idno><idno type="doi">10.1038/s41598-019-42628-6</idno><date when="2019">2019</date><idno type="wicri:Area/Pmc/Corpus">000458</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000458</idno><idno type="wicri:Area/Pmc/Curation">000458</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000458</idno><idno type="wicri:Area/Pmc/Checkpoint">000343</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000343</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies</title><author><name sortKey="Lim, Chia Chiu" sort="Lim, Chia Chiu" uniqKey="Lim C" first="Chia Chiu" last="Lim">Chia Chiu 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></author><author><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><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 Dübel</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Marks, Jd" uniqKey="Marks J">JD Marks</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ponsel, D" uniqKey="Ponsel D">D Ponsel</name></author><author><name sortKey="Neugebauer, J" uniqKey="Neugebauer J">J Neugebauer</name></author><author><name sortKey="Ladetzki Baehs, K" uniqKey="Ladetzki Baehs K">K Ladetzki-Baehs</name></author><author><name sortKey="Tissot, K" uniqKey="Tissot K">K Tissot</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Tohidkia, Mr" uniqKey="Tohidkia M">MR Tohidkia</name></author><author><name sortKey="Barar, J" uniqKey="Barar J">J Barar</name></author><author><name sortKey="Asadi, F" uniqKey="Asadi F">F Asadi</name></author><author><name sortKey="Omidi, Y" uniqKey="Omidi Y">Y Omidi</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Zhao, A" uniqKey="Zhao A">A Zhao</name></author><author><name sortKey="Tohidkia, Mr" uniqKey="Tohidkia M">MR Tohidkia</name></author><author><name sortKey="Siegel, Dl" uniqKey="Siegel D">DL Siegel</name></author><author><name sortKey="Coukos, G" uniqKey="Coukos G">G Coukos</name></author><author><name sortKey="Omidi, Y" uniqKey="Omidi Y">Y Omidi</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bornholdt, Za" uniqKey="Bornholdt Z">ZA Bornholdt</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Hoogenboom, Hr" uniqKey="Hoogenboom H">HR Hoogenboom</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Zhou, H" uniqKey="Zhou H">H Zhou</name></author><author><name sortKey="Zhang, Y L" uniqKey="Zhang Y">Y-L Zhang</name></author><author><name sortKey="Lu, G" uniqKey="Lu G">G Lu</name></author><author><name sortKey="Ji, H" uniqKey="Ji H">H Ji</name></author><author><name sortKey="Rodi, Cp" uniqKey="Rodi C">CP Rodi</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lebani, K" uniqKey="Lebani K">K Lebani</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Liu, Y" uniqKey="Liu Y">Y Liu</name></author><author><name sortKey="Yu, F" uniqKey="Yu F">F Yu</name></author><author><name sortKey="Huang, H" uniqKey="Huang H">H Huang</name></author><author><name sortKey="Han, J" uniqKey="Han J">J Han</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Viranaicken, W" uniqKey="Viranaicken W">W Viranaicken</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Boisen, Ml" uniqKey="Boisen M">ML Boisen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chew, Ch" uniqKey="Chew C">CH Chew</name></author><author><name sortKey="Lim, Yal" uniqKey="Lim Y">YAL Lim</name></author><author><name sortKey="Chua, Kh" uniqKey="Chua K">KH Chua</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Hamidon, Nh" uniqKey="Hamidon N">NH Hamidon</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Rahumatullah, A" uniqKey="Rahumatullah A">A Rahumatullah</name></author><author><name sortKey="Abdul Karim, Iz" uniqKey="Abdul Karim I">IZ Abdul Karim</name></author><author><name sortKey="Noordin, R" uniqKey="Noordin R">R Noordin</name></author><author><name sortKey="Lim, Ts" uniqKey="Lim T">TS Lim</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Gupta, Sk" uniqKey="Gupta S">SK Gupta</name></author><author><name sortKey="Shukla, P" uniqKey="Shukla P">P Shukla</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Rosano, Gl" uniqKey="Rosano G">GL Rosano</name></author><author><name sortKey="Ceccarelli, Ea" uniqKey="Ceccarelli E">EA Ceccarelli</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Costa, S" uniqKey="Costa S">S Costa</name></author><author><name sortKey="Almeida, A" uniqKey="Almeida A">A Almeida</name></author><author><name sortKey="Castro, A" uniqKey="Castro A">A Castro</name></author><author><name sortKey="Domingues, L" uniqKey="Domingues L">L Domingues</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Jia, B" uniqKey="Jia B">B Jia</name></author><author><name sortKey="Jeon, Co" uniqKey="Jeon C">CO Jeon</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Liu, Jk" uniqKey="Liu J">JK Liu</name></author></analytic></biblStruct><biblStruct></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="Barkhordarian, H" uniqKey="Barkhordarian H">H Barkhordarian</name></author><author><name sortKey="Emadi, S" uniqKey="Emadi S">S Emadi</name></author><author><name sortKey="Schulz, P" uniqKey="Schulz P">P Schulz</name></author><author><name sortKey="Sierks, Mr" uniqKey="Sierks M">MR Sierks</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bachler, Bc" uniqKey="Bachler B">BC Bachler</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Coelho, Eaf" uniqKey="Coelho E">EAF Coelho</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ding, Yl" uniqKey="Ding Y">YL Ding</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Liu, B" uniqKey="Liu B">B Liu</name></author><author><name sortKey="Huang, L" uniqKey="Huang L">L Huang</name></author><author><name sortKey="Sihlbom, C" uniqKey="Sihlbom C">C Sihlbom</name></author><author><name sortKey="Burlingame, A" uniqKey="Burlingame A">A Burlingame</name></author><author><name sortKey="Marks, Jd" uniqKey="Marks J">JD Marks</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lamboy, Ja" uniqKey="Lamboy J">JA Lamboy</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Hammers, Cm" uniqKey="Hammers C">CM Hammers</name></author><author><name sortKey="Stanley, Jr" uniqKey="Stanley J">JR Stanley</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Stausb L Gr N, B" uniqKey="Stausb L Gr N B">B Stausbøl-Grøn</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Alfaleh, Ma" uniqKey="Alfaleh M">MA Alfaleh</name></author><author><name sortKey="Jones, Ml" uniqKey="Jones M">ML Jones</name></author><author><name sortKey="Howard, Cb" uniqKey="Howard C">CB Howard</name></author><author><name sortKey="Mahler, Sm" uniqKey="Mahler S">SM Mahler</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Rajaram, K" uniqKey="Rajaram K">K Rajaram</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Shen, W" uniqKey="Shen W">W Shen</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Wu, C H" uniqKey="Wu C">C-H Wu</name></author><author><name sortKey="Liu, I J" uniqKey="Liu I">I-J Liu</name></author><author><name sortKey="Lu, R M" uniqKey="Lu R">R-M Lu</name></author><author><name sortKey="Wu, H C" uniqKey="Wu H">H-C Wu</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Adey, Nb" uniqKey="Adey N">NB Adey</name></author><author><name sortKey="Mataragnon, Ah" uniqKey="Mataragnon A">AH Mataragnon</name></author><author><name sortKey="Rider, Je" uniqKey="Rider J">JE Rider</name></author><author><name sortKey="Carter, Jm" uniqKey="Carter J">JM Carter</name></author><author><name sortKey="Kay, Bk" uniqKey="Kay B">BK Kay</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bazan, J" uniqKey="Bazan J">J Bazan</name></author><author><name sortKey="Calkosi Ski, I" uniqKey="Calkosi Ski I">I Całkosiński</name></author><author><name sortKey="Gamian, A" uniqKey="Gamian A">A Gamian</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Hust, M" uniqKey="Hust M">M Hust</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Venet, S" uniqKey="Venet S">S Venet</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bradbury, Ar" uniqKey="Bradbury A">AR Bradbury</name></author><author><name sortKey="Sidhu, S" uniqKey="Sidhu S">S Sidhu</name></author><author><name sortKey="Dubel, S" uniqKey="Dubel S">S Dübel</name></author><author><name sortKey="Mccafferty, J" uniqKey="Mccafferty J">J McCafferty</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Townsend, Cl" uniqKey="Townsend C">CL Townsend</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Perfetti, V" uniqKey="Perfetti V">V Perfetti</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Smith, K" uniqKey="Smith K">K Smith</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Guan, Y" uniqKey="Guan Y">Y Guan</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="De Boer, E" uniqKey="De Boer E">E de Boer</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Humbert, N" uniqKey="Humbert N">N Humbert</name></author><author><name sortKey="Zocchi, A" uniqKey="Zocchi A">A Zocchi</name></author><author><name sortKey="Ward, Tr" uniqKey="Ward T">TR Ward</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Li, Y" uniqKey="Li Y">Y Li</name></author><author><name sortKey="Sousa, R" uniqKey="Sousa R">R Sousa</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Sorenson, Ae" uniqKey="Sorenson A">AE Sorenson</name></author><author><name sortKey="Askin, Sp" uniqKey="Askin S">SP Askin</name></author><author><name sortKey="Schaeffer, Pm" uniqKey="Schaeffer P">PM Schaeffer</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Fairhead, M" uniqKey="Fairhead M">M Fairhead</name></author><author><name sortKey="Howarth, M" uniqKey="Howarth M">M Howarth</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chen, Y" uniqKey="Chen Y">Y Chen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Jiang, L" uniqKey="Jiang L">L Jiang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ying, T" uniqKey="Ying T">T Ying</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wang, L" uniqKey="Wang L">L Wang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Yamaoka, Y" uniqKey="Yamaoka Y">Y Yamaoka</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ying, T" uniqKey="Ying T">T Ying</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kugler, J" uniqKey="Kugler J">J Kügler</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ismail, Nf" uniqKey="Ismail N">NF Ismail</name></author><author><name sortKey="Lim, Ts" uniqKey="Lim T">TS Lim</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Lim, Bn" uniqKey="Lim B">BN Lim</name></author><author><name sortKey="Chin, Cf" uniqKey="Chin C">CF Chin</name></author><author><name sortKey="Choong, Ys" uniqKey="Choong Y">YS Choong</name></author><author><name sortKey="Ismail, A" uniqKey="Ismail A">A Ismail</name></author><author><name sortKey="Lim, Ts" uniqKey="Lim T">TS Lim</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Brochet, X" uniqKey="Brochet X">X Brochet</name></author><author><name sortKey="Lefranc, M P" uniqKey="Lefranc M">M-P Lefranc</name></author><author><name sortKey="Giudicelli, V" uniqKey="Giudicelli V">V Giudicelli</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="De Kruif, J" uniqKey="De Kruif J">J de Kruif</name></author><author><name sortKey="Terstappen, L" uniqKey="Terstappen L">L Terstappen</name></author><author><name sortKey="Boel, E" uniqKey="Boel E">E Boel</name></author><author><name sortKey="Logtenberg, T" uniqKey="Logtenberg T">T Logtenberg</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Giordano, Rj" uniqKey="Giordano R">RJ Giordano</name></author><author><name sortKey="Card Vila, M" uniqKey="Card Vila M">M Cardó-Vila</name></author><author><name sortKey="Lahdenranta, J" uniqKey="Lahdenranta J">J Lahdenranta</name></author><author><name sortKey="Pasqualini, R" uniqKey="Pasqualini R">R Pasqualini</name></author><author><name sortKey="Arap, W" uniqKey="Arap W">W Arap</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Eisenhardt, Su" uniqKey="Eisenhardt S">SU Eisenhardt</name></author><author><name sortKey="Schwarz, M" uniqKey="Schwarz M">M Schwarz</name></author><author><name sortKey="Bassler, N" uniqKey="Bassler N">N Bassler</name></author><author><name sortKey="Peter, K" uniqKey="Peter K">K Peter</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="S Rensen, Md" uniqKey="S Rensen M">MD Sørensen</name></author><author><name sortKey="Agerholm, Ie" uniqKey="Agerholm I">IE Agerholm</name></author><author><name sortKey="Christensen, B" 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><pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Sci Rep</journal-id><journal-id journal-id-type="iso-abbrev">Sci Rep</journal-id><journal-title-group><journal-title>Scientific Reports</journal-title></journal-title-group><issn pub-type="epub">2045-2322</issn><publisher><publisher-name>Nature Publishing Group UK</publisher-name><publisher-loc>London</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">30988390</article-id><article-id pub-id-type="pmc">6465254</article-id><article-id pub-id-type="publisher-id">42628</article-id><article-id pub-id-type="doi">10.1038/s41598-019-42628-6</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Lim</surname><given-names>Chia Chiu</given-names></name><xref ref-type="aff" rid="Aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Woo</surname><given-names>Patrick C. Y.</given-names></name><xref ref-type="aff" rid="Aff2">2</xref></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">http://orcid.org/0000-0002-0656-3045</contrib-id><name><surname>Lim</surname><given-names>Theam Soon</given-names></name><address><email>theamsoon@usm.my</email></address><xref ref-type="aff" rid="Aff1">1</xref><xref ref-type="aff" rid="Aff3">3</xref></contrib><aff id="Aff1"><label>1</label><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</aff><aff id="Aff2"><label>2</label><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</aff><aff id="Aff3"><label>3</label><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</aff></contrib-group><pub-date pub-type="epub"><day>15</day><month>4</month><year>2019</year></pub-date><pub-date pub-type="pmc-release"><day>15</day><month>4</month><year>2019</year></pub-date><pub-date pub-type="collection"><year>2019</year></pub-date><volume>9</volume><elocation-id>6088</elocation-id><history><date date-type="received"><day>16</day><month>7</month><year>2018</year></date><date date-type="accepted"><day>4</day><month>4</month><year>2019</year></date></history><permissions><copyright-statement>© The Author(s) 2019</copyright-statement><license license-type="OpenAccess"><license-p><bold>Open Access</bold> This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">http://creativecommons.org/licenses/by/4.0/</ext-link>.</license-p></license></permissions><abstract id="Abs1"><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></abstract><kwd-group kwd-group-type="npg-subject"><title>Subject terms</title><kwd>Infectious diseases</kwd><kwd>Bacteriophages</kwd><kwd>Molecular medicine</kwd></kwd-group><funding-group><award-group><funding-source><institution>Theme-based Research Scheme (Project No. T11/707/15), University Grant Committee, Hong Kong</institution></funding-source></award-group></funding-group><funding-group><award-group><funding-source><institution>Malaysian Ministry of Education through the Fundamental Research Grant Scheme (203/CIPPM/6711658)</institution></funding-source></award-group></funding-group><custom-meta-group><custom-meta><meta-name>issue-copyright-statement</meta-name><meta-value>© The Author(s) 2019</meta-value></custom-meta></custom-meta-group></article-meta></front></pmc><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)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Pmc/Checkpoint
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000343 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Pmc/Checkpoint/biblio.hfd -nk 000343 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= MersV1
|flux= Pmc
|étape= Checkpoint
|type= RBID
|clé= PMC:6465254
|texte= Development of a Phage Display Panning Strategy Utilizing Crude Antigens: Isolation of MERS-CoV Nucleoprotein human antibodies
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Pmc/Checkpoint/RBID.i -Sk "pubmed:30988390" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Pmc/Checkpoint/biblio.hfd \
| NlmPubMed2Wicri -a MersV1
| This area was generated with Dilib version V0.6.33. |  |