Development of a platform process for the production and purification of single‐domain antibodies
Single-domain antibodies (sdAbs) offer the affinity and therapeutic value of conventional antibodies, with increased stability and solubility. Unlike conventional antibodies, however, sdAbs do not benefit from a platform manufacturing process. While successful production of a variety of sdAbs has be...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | info:ar-repo/semantics/artículo |
| Language: | Inglés |
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Wiley
2023
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| Online Access: | http://hdl.handle.net/20.500.12123/15527 https://onlinelibrary.wiley.com/doi/10.1002/bit.27724 https://doi.org/10.1002/bit.27724 |
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| author | Crowell, Laura E. Goodwine, Chaz Sosa Holt, Carla Solange Rocha, Lucía Alejandra Vega, Celina Guadalupe Rodriguez-Aponte, Sergio A. Dalvie, Neil C. Tracey, Mary Kate Puntel, Mariana Wigdorovitz, Andres Parreño, Gladys Love, Kerry R. Cramer, Steven M. Love, J. Christopher |
| author_browse | Cramer, Steven M. Crowell, Laura E. Dalvie, Neil C. Goodwine, Chaz Love, J. Christopher Love, Kerry R. Parreño, Gladys Puntel, Mariana Rocha, Lucía Alejandra Rodriguez-Aponte, Sergio A. Sosa Holt, Carla Solange Tracey, Mary Kate Vega, Celina Guadalupe Wigdorovitz, Andres |
| author_facet | Crowell, Laura E. Goodwine, Chaz Sosa Holt, Carla Solange Rocha, Lucía Alejandra Vega, Celina Guadalupe Rodriguez-Aponte, Sergio A. Dalvie, Neil C. Tracey, Mary Kate Puntel, Mariana Wigdorovitz, Andres Parreño, Gladys Love, Kerry R. Cramer, Steven M. Love, J. Christopher |
| author_sort | Crowell, Laura E. |
| collection | INTA Digital |
| description | Single-domain antibodies (sdAbs) offer the affinity and therapeutic value of conventional antibodies, with increased stability and solubility. Unlike conventional antibodies, however, sdAbs do not benefit from a platform manufacturing process. While successful production of a variety of sdAbs has been shown in numerous hosts, purification methods are often molecule specific or require affinity tags, which generally cannot be used in clinical manufacturing due to regulatory concerns. Here, we have developed a broadly applicable production and purification process for sdAbs in Komagataella phaffii (Pichia pastoris) and demonstrated the production of eight different sdAbs at a quality appropriate for nonclinical studies. We developed a two-step, integrated purification process without the use of affinity resins and showed that modification of a single process parameter, pH of the bridging buffer, was required for the successful purification of a variety of sdAbs. Further, we determined that this parameter can be predicted based only on the biophysical characteristics of the target molecule. Using these methods, we produced nonclinical quality sdAbs as few as 5 weeks after identifying the product sequence. Nonclinical studies of three different sdAbs showed that molecules produced using our platform process conferred protection against viral shedding of rotavirus or H1N1 influenza and were equivalent to similar molecules produced in Escherichia coli and purified using affinity tags. |
| format | info:ar-repo/semantics/artículo |
| id | INTA15527 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2023 |
| publishDateRange | 2023 |
| publishDateSort | 2023 |
| publisher | Wiley |
| publisherStr | Wiley |
| record_format | dspace |
| spelling | INTA155272023-10-11T16:06:09Z Development of a platform process for the production and purification of single‐domain antibodies Crowell, Laura E. Goodwine, Chaz Sosa Holt, Carla Solange Rocha, Lucía Alejandra Vega, Celina Guadalupe Rodriguez-Aponte, Sergio A. Dalvie, Neil C. Tracey, Mary Kate Puntel, Mariana Wigdorovitz, Andres Parreño, Gladys Love, Kerry R. Cramer, Steven M. Love, J. Christopher Pichia pastoris Purificación Anticuerpos Purification Antibodies Komagataella phaffii Single-domain antibodies (sdAbs) offer the affinity and therapeutic value of conventional antibodies, with increased stability and solubility. Unlike conventional antibodies, however, sdAbs do not benefit from a platform manufacturing process. While successful production of a variety of sdAbs has been shown in numerous hosts, purification methods are often molecule specific or require affinity tags, which generally cannot be used in clinical manufacturing due to regulatory concerns. Here, we have developed a broadly applicable production and purification process for sdAbs in Komagataella phaffii (Pichia pastoris) and demonstrated the production of eight different sdAbs at a quality appropriate for nonclinical studies. We developed a two-step, integrated purification process without the use of affinity resins and showed that modification of a single process parameter, pH of the bridging buffer, was required for the successful purification of a variety of sdAbs. Further, we determined that this parameter can be predicted based only on the biophysical characteristics of the target molecule. Using these methods, we produced nonclinical quality sdAbs as few as 5 weeks after identifying the product sequence. Nonclinical studies of three different sdAbs showed that molecules produced using our platform process conferred protection against viral shedding of rotavirus or H1N1 influenza and were equivalent to similar molecules produced in Escherichia coli and purified using affinity tags. Instituto de Virología Fil: Crowell, Laura E. Massachusetts Institute of Technology. The Koch Institute for Integrative Cancer Research; Estados Unidos Fil: Crowell, Laura E. Massachusetts Institute of Technology. Department of Chemical Engineering; Estados Unidos Fil: Goodwine, Chaz. Rensselaer Polytechnic Institute. Department of Chemical and Biological Engineering; Estados Unidos Fil: Goodwine, Chaz. Rensselaer Polytechnic Institute. Center for Biotechnology and Interdisciplinary Studies; Estados Unidos Fil: Sosa Holt, Carla Solange. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina Fil: Rocha, Lucía Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina Fil: Vega, Celina Guadalupe. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina Fil: Rodriguez-Aponte, Sergio A. Massachusetts Institute of Technology. The Koch Institute for Integrative Cancer Research; Estados Unidos Fil: Rodriguez-Aponte, Sergio A. Massachusetts Institute of Technology. Department of Biological Engineering; Estados Unidos Fil: Dalvie, Neil C. Massachusetts Institute of Technology. The Koch Institute for Integrative Cancer Research; Estados Unidos Fil: Dalvie, Neil C. Massachusetts Institute of Technology. Department of Chemical Engineering; Estados Unidos Fil: Tracey, Mary Kate. Massachusetts Institute of Technology. The Koch Institute for Integrative Cancer Research; Estados Unidos Fil: Puntel, Mariana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina Fil: Wigdorovitz, Andres. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina Fil: Parreño, Gladys Viviana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina Fil: Love, Kerry R. Massachusetts Institute of Technology. The Koch Institute for Integrative Cancer Research; Estados Unidos Fil: Cramer, Steven M. Rensselaer Polytechnic Institute. Department of Chemical and Biological Engineering; Estados Unidos Fil: Cramer, Steven M. Rensselaer Polytechnic Institute. Center for Biotechnology and Interdisciplinary Studies; Estados Unidos Fil: Love, J. Christopher. Massachusetts Institute of Technology. The Koch Institute for Integrative Cancer Research; Estados Unidos Fil: Love, J. Christopher. Massachusetts Institute of Technology. Department of Chemical Engineering; Estados Unidos 2023-10-11T16:01:58Z 2023-10-11T16:01:58Z 2021-09 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/15527 https://onlinelibrary.wiley.com/doi/10.1002/bit.27724 1097-0290 https://doi.org/10.1002/bit.27724 eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Wiley Biotechnology and Bioengineering 118 (9) : 3348-3358. (September 2021) |
| spellingShingle | Pichia pastoris Purificación Anticuerpos Purification Antibodies Komagataella phaffii Crowell, Laura E. Goodwine, Chaz Sosa Holt, Carla Solange Rocha, Lucía Alejandra Vega, Celina Guadalupe Rodriguez-Aponte, Sergio A. Dalvie, Neil C. Tracey, Mary Kate Puntel, Mariana Wigdorovitz, Andres Parreño, Gladys Love, Kerry R. Cramer, Steven M. Love, J. Christopher Development of a platform process for the production and purification of single‐domain antibodies |
| title | Development of a platform process for the production and purification of single‐domain antibodies |
| title_full | Development of a platform process for the production and purification of single‐domain antibodies |
| title_fullStr | Development of a platform process for the production and purification of single‐domain antibodies |
| title_full_unstemmed | Development of a platform process for the production and purification of single‐domain antibodies |
| title_short | Development of a platform process for the production and purification of single‐domain antibodies |
| title_sort | development of a platform process for the production and purification of single domain antibodies |
| topic | Pichia pastoris Purificación Anticuerpos Purification Antibodies Komagataella phaffii |
| url | http://hdl.handle.net/20.500.12123/15527 https://onlinelibrary.wiley.com/doi/10.1002/bit.27724 https://doi.org/10.1002/bit.27724 |
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