Development of a platform process for the production and purification of single-domain antibodies

Autores
Crowell, Laura E.; Goodwine, Chaz; Sosa Holt, Carla Solange; Rocha, Lucia; Vega, Celina Guadalupe; Rodriguez, Sergio A.; Dalvie, Neil C.; Tracey, Mary K.; Puntel, Mariana; Wigdorovitz, Andrés; Parreño, Gladys Viviana; Love, Kerry R.; Cramer, Steven M.; Love, J. Christopher
Año de publicación
2021
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
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.
Fil: Crowell, Laura E.. Massachusetts Institute of Technology; Estados Unidos
Fil: Goodwine, Chaz. Rensselaer Polytechnic Institute; Estados Unidos
Fil: Sosa Holt, Carla Solange. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Rocha, Lucia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Vega, Celina Guadalupe. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología e Innovaciones Tecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Virología e Innovaciones Tecnológicas; Argentina
Fil: Rodriguez, Sergio A.. Massachusetts Institute of Technology; Estados Unidos
Fil: Dalvie, Neil C.. Massachusetts Institute of Technology; Estados Unidos
Fil: Tracey, Mary K.. Massachusetts Institute of Technology; Estados Unidos
Fil: Puntel, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Wigdorovitz, Andrés. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Parreño, Gladys Viviana. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología e Innovaciones Tecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Virología e Innovaciones Tecnológicas; Argentina
Fil: Love, Kerry R.. Massachusetts Institute of Technology; Estados Unidos
Fil: Cramer, Steven M.. Rensselaer Polytechnic Institute; Estados Unidos
Fil: Love, J. Christopher. Massachusetts Institute of Technology; Estados Unidos
Materia
INTEGRATED PURIFICATION
KOMAGATAELLA PHAFFII
PICHIA PASTORIS
SINGLE-DOMAIN ANTIBODIES
STRAIGHT-THROUGH CHROMATOGRAPHY
VHH
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/185008
Acceder
id CONICETDig_d7845b51f1fd66c4c1c5837b5d6e29ec
oai_identifier_str oai:ri.conicet.gov.ar:11336/185008
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Development of a platform process for the production and purification of single-domain antibodiesCrowell, Laura E.Goodwine, ChazSosa Holt, Carla SolangeRocha, LuciaVega, Celina GuadalupeRodriguez, Sergio A.Dalvie, Neil C.Tracey, Mary K.Puntel, MarianaWigdorovitz, AndrésParreño, Gladys VivianaLove, Kerry R.Cramer, Steven M.Love, J. ChristopherINTEGRATED PURIFICATIONKOMAGATAELLA PHAFFIIPICHIA PASTORISSINGLE-DOMAIN ANTIBODIESSTRAIGHT-THROUGH CHROMATOGRAPHYVHHhttps://purl.org/becyt/ford/2.10https://purl.org/becyt/ford/2Single-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.Fil: Crowell, Laura E.. Massachusetts Institute of Technology; Estados UnidosFil: Goodwine, Chaz. Rensselaer Polytechnic Institute; Estados UnidosFil: Sosa Holt, Carla Solange. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; ArgentinaFil: Rocha, Lucia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; ArgentinaFil: Vega, Celina Guadalupe. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología e Innovaciones Tecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Virología e Innovaciones Tecnológicas; ArgentinaFil: Rodriguez, Sergio A.. Massachusetts Institute of Technology; Estados UnidosFil: Dalvie, Neil C.. Massachusetts Institute of Technology; Estados UnidosFil: Tracey, Mary K.. Massachusetts Institute of Technology; Estados UnidosFil: Puntel, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; ArgentinaFil: Wigdorovitz, Andrés. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; ArgentinaFil: Parreño, Gladys Viviana. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología e Innovaciones Tecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Virología e Innovaciones Tecnológicas; ArgentinaFil: Love, Kerry R.. Massachusetts Institute of Technology; Estados UnidosFil: Cramer, Steven M.. Rensselaer Polytechnic Institute; Estados UnidosFil: Love, J. Christopher. Massachusetts Institute of Technology; Estados UnidosJohn Wiley & Sons2021-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/185008Crowell, Laura E.; Goodwine, Chaz; Sosa Holt, Carla Solange; Rocha, Lucia; Vega, Celina Guadalupe; et al.; Development of a platform process for the production and purification of single-domain antibodies; John Wiley & Sons; Bioengineering And Biotechnology; 118; 9; 2-2021; 3348-33580006-3592CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/10.1002/bit.27724info:eu-repo/semantics/altIdentifier/doi/10.1002/bit.27724info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:25:08Zoai:ri.conicet.gov.ar:11336/185008instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-10 13:25:08.992CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv 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
spellingShingle Development of a platform process for the production and purification of single-domain antibodies
Crowell, Laura E.
INTEGRATED PURIFICATION
KOMAGATAELLA PHAFFII
PICHIA PASTORIS
SINGLE-DOMAIN ANTIBODIES
STRAIGHT-THROUGH CHROMATOGRAPHY
VHH
title_short 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_sort Development of a platform process for the production and purification of single-domain antibodies
dc.creator.none.fl_str_mv Crowell, Laura E.
Goodwine, Chaz
Sosa Holt, Carla Solange
Rocha, Lucia
Vega, Celina Guadalupe
Rodriguez, Sergio A.
Dalvie, Neil C.
Tracey, Mary K.
Puntel, Mariana
Wigdorovitz, Andrés
Parreño, Gladys Viviana
Love, Kerry R.
Cramer, Steven M.
Love, J. Christopher
author Crowell, Laura E.
author_facet Crowell, Laura E.
Goodwine, Chaz
Sosa Holt, Carla Solange
Rocha, Lucia
Vega, Celina Guadalupe
Rodriguez, Sergio A.
Dalvie, Neil C.
Tracey, Mary K.
Puntel, Mariana
Wigdorovitz, Andrés
Parreño, Gladys Viviana
Love, Kerry R.
Cramer, Steven M.
Love, J. Christopher
author_role author
author2 Goodwine, Chaz
Sosa Holt, Carla Solange
Rocha, Lucia
Vega, Celina Guadalupe
Rodriguez, Sergio A.
Dalvie, Neil C.
Tracey, Mary K.
Puntel, Mariana
Wigdorovitz, Andrés
Parreño, Gladys Viviana
Love, Kerry R.
Cramer, Steven M.
Love, J. Christopher
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv INTEGRATED PURIFICATION
KOMAGATAELLA PHAFFII
PICHIA PASTORIS
SINGLE-DOMAIN ANTIBODIES
STRAIGHT-THROUGH CHROMATOGRAPHY
VHH
topic INTEGRATED PURIFICATION
KOMAGATAELLA PHAFFII
PICHIA PASTORIS
SINGLE-DOMAIN ANTIBODIES
STRAIGHT-THROUGH CHROMATOGRAPHY
VHH
purl_subject.fl_str_mv https://purl.org/becyt/ford/2.10
https://purl.org/becyt/ford/2
dc.description.none.fl_txt_mv 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.
Fil: Crowell, Laura E.. Massachusetts Institute of Technology; Estados Unidos
Fil: Goodwine, Chaz. Rensselaer Polytechnic Institute; Estados Unidos
Fil: Sosa Holt, Carla Solange. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Rocha, Lucia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Vega, Celina Guadalupe. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología e Innovaciones Tecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Virología e Innovaciones Tecnológicas; Argentina
Fil: Rodriguez, Sergio A.. Massachusetts Institute of Technology; Estados Unidos
Fil: Dalvie, Neil C.. Massachusetts Institute of Technology; Estados Unidos
Fil: Tracey, Mary K.. Massachusetts Institute of Technology; Estados Unidos
Fil: Puntel, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Wigdorovitz, Andrés. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Parreño, Gladys Viviana. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología e Innovaciones Tecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Virología e Innovaciones Tecnológicas; Argentina
Fil: Love, Kerry R.. Massachusetts Institute of Technology; Estados Unidos
Fil: Cramer, Steven M.. Rensselaer Polytechnic Institute; Estados Unidos
Fil: Love, J. Christopher. Massachusetts Institute of Technology; Estados Unidos
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.
publishDate 2021
dc.date.none.fl_str_mv 2021-02
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/185008
Crowell, Laura E.; Goodwine, Chaz; Sosa Holt, Carla Solange; Rocha, Lucia; Vega, Celina Guadalupe; et al.; Development of a platform process for the production and purification of single-domain antibodies; John Wiley & Sons; Bioengineering And Biotechnology; 118; 9; 2-2021; 3348-3358
0006-3592
CONICET Digital
CONICET
url http://hdl.handle.net/11336/185008
identifier_str_mv Crowell, Laura E.; Goodwine, Chaz; Sosa Holt, Carla Solange; Rocha, Lucia; Vega, Celina Guadalupe; et al.; Development of a platform process for the production and purification of single-domain antibodies; John Wiley & Sons; Bioengineering And Biotechnology; 118; 9; 2-2021; 3348-3358
0006-3592
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/10.1002/bit.27724
info:eu-repo/semantics/altIdentifier/doi/10.1002/bit.27724
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv John Wiley & Sons
publisher.none.fl_str_mv John Wiley & Sons
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 12.48226

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