Automated pipeline for rapid production and screening of HIV-specific monoclonal antibodies using pichia pastoris
Monoclonal antibodies (mAbs) that bind and neutralize human pathogens have great therapeutic potential. Advances in automated screening and liquid handling have resulted in the ability to discover antigen-specific antibodies either directly from human blood or from various combinatorial libraries (...
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| Format: | Article |
| Language: | en_US |
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Wiley Blackwell
2017
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| Online Access: | http://hdl.handle.net/1721.1/109438 https://orcid.org/0000-0003-0921-3144 |
| _version_ | 1826196973433126912 |
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| author | Shah, Kartik A Clark, John J Goods, Brittany A. Politano, Timothy James Mozdzierz, Nicholas Joseph Zimnisky, Ross Leeson, Rachel Love, John C Love, Kerry R. |
| author2 | Massachusetts Institute of Technology. Center for Biomedical Innovation |
| author_facet | Massachusetts Institute of Technology. Center for Biomedical Innovation Shah, Kartik A Clark, John J Goods, Brittany A. Politano, Timothy James Mozdzierz, Nicholas Joseph Zimnisky, Ross Leeson, Rachel Love, John C Love, Kerry R. |
| author_sort | Shah, Kartik A |
| collection | MIT |
| description | Monoclonal antibodies (mAbs) that bind and neutralize human pathogens have great therapeutic potential. Advances in automated screening and liquid handling have resulted in the ability to discover antigen-specific antibodies either directly from human blood or from various
combinatorial libraries (phage, bacteria or yeast). There remain, however, bottlenecks in the cloning, expression and evaluation of such lead antibodies identified in primary screens that hinder high-throughput screening. As such, ‘hit-to-lead identification’ remains both expensive and
time-consuming. By combining the advantages of overlap extension PCR (OE-PCR) and a genetically stable yet easily manipulatable microbial expression host Pichia pastoris, we have developed an automated pipeline for the rapid production and screening of full-length antigenspecific
mAbs. Here, we demonstrate the speed, feasibility and cost-effectiveness of our approach by generating several broadly neutralizing antibodies against human immunodeficiency virus (HIV). |
| first_indexed | 2024-09-23T10:40:28Z |
| format | Article |
| id | mit-1721.1/109438 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T10:40:28Z |
| publishDate | 2017 |
| publisher | Wiley Blackwell |
| record_format | dspace |
| spelling | mit-1721.1/1094382022-09-27T14:12:47Z Automated pipeline for rapid production and screening of HIV-specific monoclonal antibodies using pichia pastoris Shah, Kartik A Clark, John J Goods, Brittany A. Politano, Timothy James Mozdzierz, Nicholas Joseph Zimnisky, Ross Leeson, Rachel Love, John C Love, Kerry R. Massachusetts Institute of Technology. Center for Biomedical Innovation Massachusetts Institute of Technology. Department of Chemical Engineering Koch Institute for Integrative Cancer Research at MIT Shah, Kartik A Clark, John J Goods, Brittany A. Politano, Timothy James Mozdzierz, Nicholas Joseph Zimnisky, Ross Leeson, Rachel Love, John C Love, Kerry R. Monoclonal antibodies (mAbs) that bind and neutralize human pathogens have great therapeutic potential. Advances in automated screening and liquid handling have resulted in the ability to discover antigen-specific antibodies either directly from human blood or from various combinatorial libraries (phage, bacteria or yeast). There remain, however, bottlenecks in the cloning, expression and evaluation of such lead antibodies identified in primary screens that hinder high-throughput screening. As such, ‘hit-to-lead identification’ remains both expensive and time-consuming. By combining the advantages of overlap extension PCR (OE-PCR) and a genetically stable yet easily manipulatable microbial expression host Pichia pastoris, we have developed an automated pipeline for the rapid production and screening of full-length antigenspecific mAbs. Here, we demonstrate the speed, feasibility and cost-effectiveness of our approach by generating several broadly neutralizing antibodies against human immunodeficiency virus (HIV). Bill & Melinda Gates Foundation United States. Defense Advanced Research Projects Agency Space and Naval Warfare Systems Center San Diego (U.S.) (Contract N66001-13-C-4025) W. M. Keck Foundation National Institute of Allergy and Infectious Diseases (U.S.) (U19AI090970). National Cancer Institute (U.S.) (David H. Koch Institute for Integrative Cancer Research at MIT. Support (Core) Grant P30-CA14051) 2017-05-30T20:04:48Z 2017-05-30T20:04:48Z 2015-07 2015-05 Article http://purl.org/eprint/type/JournalArticle 0006-3592 http://hdl.handle.net/1721.1/109438 Shah, Kartik A. et al. “Automated Pipeline for Rapid Production and Screening of HIV-Specific Monoclonal Antibodies Using Pichia Pastoris: Automated Pipeline for Rapid Production and Screening.” Biotechnology and Bioengineering 112.12 (2015): 2624–2629. https://orcid.org/0000-0003-0921-3144 en_US http://dx.doi.org/10.1002/bit.25663 Biotechnology and Bioengineering Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf Wiley Blackwell PMC |
| spellingShingle | Shah, Kartik A Clark, John J Goods, Brittany A. Politano, Timothy James Mozdzierz, Nicholas Joseph Zimnisky, Ross Leeson, Rachel Love, John C Love, Kerry R. Automated pipeline for rapid production and screening of HIV-specific monoclonal antibodies using pichia pastoris |
| title | Automated pipeline for rapid production and screening of HIV-specific monoclonal antibodies using pichia pastoris |
| title_full | Automated pipeline for rapid production and screening of HIV-specific monoclonal antibodies using pichia pastoris |
| title_fullStr | Automated pipeline for rapid production and screening of HIV-specific monoclonal antibodies using pichia pastoris |
| title_full_unstemmed | Automated pipeline for rapid production and screening of HIV-specific monoclonal antibodies using pichia pastoris |
| title_short | Automated pipeline for rapid production and screening of HIV-specific monoclonal antibodies using pichia pastoris |
| title_sort | automated pipeline for rapid production and screening of hiv specific monoclonal antibodies using pichia pastoris |
| url | http://hdl.handle.net/1721.1/109438 https://orcid.org/0000-0003-0921-3144 |
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