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Resumen
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
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dc.contributor.author | Crowell, Laura E. | |
dc.contributor.author | Goodwine, Chaz | |
dc.contributor.author | Sosa Holt, Carla Solange | |
dc.contributor.author | Rocha, Lucía Alejandra | |
dc.contributor.author | Vega, Celina Guadalupe | |
dc.contributor.author | Rodriguez-Aponte, Sergio A. | |
dc.contributor.author | Dalvie, Neil C. | |
dc.contributor.author | Tracey, Mary Kate | |
dc.contributor.author | Puntel, Mariana | |
dc.contributor.author | Wigdorovitz, Andres | |
dc.contributor.author | Parreño, Gladys | |
dc.contributor.author | Love, Kerry R. | |
dc.contributor.author | Cramer, Steven M. | |
dc.contributor.author | Love, J. Christopher | |
dc.date.accessioned | 2023-10-11T16:01:58Z | |
dc.date.available | 2023-10-11T16:01:58Z | |
dc.date.issued | 2021-09 | |
dc.identifier.issn | 1097-0290 | |
dc.identifier.other | https://doi.org/10.1002/bit.27724 | |
dc.identifier.uri | http://hdl.handle.net/20.500.12123/15527 | |
dc.identifier.uri | https://onlinelibrary.wiley.com/doi/10.1002/bit.27724 | |
dc.description.abstract | 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. | eng |
dc.format | application/pdf | es_AR |
dc.language.iso | eng | es_AR |
dc.publisher | Wiley | es_AR |
dc.rights | info:eu-repo/semantics/openAccess | es_AR |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-sa/4.0/ | es_AR |
dc.source | Biotechnology and Bioengineering 118 (9) : 3348-3358. (September 2021) | es_AR |
dc.subject | Pichia pastoris | es_AR |
dc.subject | Purification | eng |
dc.subject | Purificación | es_AR |
dc.subject | Antibodies | eng |
dc.subject | Anticuerpos | es_AR |
dc.subject.other | Komagataella phaffii | es_AR |
dc.title | Development of a platform process for the production and purification of single‐domain antibodies | es_AR |
dc.type | info:ar-repo/semantics/artículo | es_AR |
dc.type | info:eu-repo/semantics/article | es_AR |
dc.type | info:eu-repo/semantics/publishedVersion | es_AR |
dc.rights.license | Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) | es_AR |
dc.description.origen | Instituto de Virología | es_AR |
dc.description.fil | Fil: Crowell, Laura E. Massachusetts Institute of Technology. The Koch Institute for Integrative Cancer Research; Estados Unidos | es_AR |
dc.description.fil | Fil: Crowell, Laura E. Massachusetts Institute of Technology. Department of Chemical Engineering; Estados Unidos | es_AR |
dc.description.fil | Fil: Goodwine, Chaz. Rensselaer Polytechnic Institute. Department of Chemical and Biological Engineering; Estados Unidos | es_AR |
dc.description.fil | Fil: Goodwine, Chaz. Rensselaer Polytechnic Institute. Center for Biotechnology and Interdisciplinary Studies; Estados Unidos | es_AR |
dc.description.fil | Fil: Sosa Holt, Carla Solange. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina | es_AR |
dc.description.fil | Fil: Rocha, Lucía Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina | es_AR |
dc.description.fil | Fil: Vega, Celina Guadalupe. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina | es_AR |
dc.description.fil | Fil: Rodriguez-Aponte, Sergio A. Massachusetts Institute of Technology. The Koch Institute for Integrative Cancer Research; Estados Unidos | es_AR |
dc.description.fil | Fil: Rodriguez-Aponte, Sergio A. Massachusetts Institute of Technology. Department of Biological Engineering; Estados Unidos | es_AR |
dc.description.fil | Fil: Dalvie, Neil C. Massachusetts Institute of Technology. The Koch Institute for Integrative Cancer Research; Estados Unidos | es_AR |
dc.description.fil | Fil: Dalvie, Neil C. Massachusetts Institute of Technology. Department of Chemical Engineering; Estados Unidos | es_AR |
dc.description.fil | Fil: Tracey, Mary Kate. Massachusetts Institute of Technology. The Koch Institute for Integrative Cancer Research; Estados Unidos | es_AR |
dc.description.fil | Fil: Puntel, Mariana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina | es_AR |
dc.description.fil | Fil: Wigdorovitz, Andres. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina | es_AR |
dc.description.fil | Fil: Parreño, Gladys Viviana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina | es_AR |
dc.description.fil | Fil: Love, Kerry R. Massachusetts Institute of Technology. The Koch Institute for Integrative Cancer Research; Estados Unidos | es_AR |
dc.description.fil | Fil: Cramer, Steven M. Rensselaer Polytechnic Institute. Department of Chemical and Biological Engineering; Estados Unidos | es_AR |
dc.description.fil | Fil: Cramer, Steven M. Rensselaer Polytechnic Institute. Center for Biotechnology and Interdisciplinary Studies; Estados Unidos | es_AR |
dc.description.fil | Fil: Love, J. Christopher. Massachusetts Institute of Technology. The Koch Institute for Integrative Cancer Research; Estados Unidos | es_AR |
dc.description.fil | Fil: Love, J. Christopher. Massachusetts Institute of Technology. Department of Chemical Engineering; Estados Unidos | es_AR |
dc.subtype | cientifico |
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