Cargando…
Novel Antibody Drug Conjugates Targeting Tumor-Associated Receptor Tyrosine Kinase ROR2 by Functional Screening of Fully Human Antibody Libraries Using Transpo-mAb Display on Progenitor B Cells
Receptor tyrosine kinase-like orphan receptor 2 (ROR2) has been identified as a highly relevant tumor-associated antigen in a variety of cancer indications of high unmet medical need, including renal cell carcinoma and osteosarcoma, making it an attractive target for targeted cancer therapy. Here, w...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6224377/ https://www.ncbi.nlm.nih.gov/pubmed/30450096 http://dx.doi.org/10.3389/fimmu.2018.02490 |
_version_ | 1783369585294049280 |
---|---|
author | Hellmann, Ina Waldmeier, Lorenz Bannwarth-Escher, Marie-Christine Maslova, Kseniya Wolter, Fabian I. Grawunder, Ulf Beerli, Roger R. |
author_facet | Hellmann, Ina Waldmeier, Lorenz Bannwarth-Escher, Marie-Christine Maslova, Kseniya Wolter, Fabian I. Grawunder, Ulf Beerli, Roger R. |
author_sort | Hellmann, Ina |
collection | PubMed |
description | Receptor tyrosine kinase-like orphan receptor 2 (ROR2) has been identified as a highly relevant tumor-associated antigen in a variety of cancer indications of high unmet medical need, including renal cell carcinoma and osteosarcoma, making it an attractive target for targeted cancer therapy. Here, we describe the de novo discovery of fully human ROR2-specific antibodies and potent antibody drug conjugates (ADCs) derived thereof by combining antibody discovery from immune libraries of human immunoglobulin transgenic animals using the Transpo-mAb mammalian cell-based IgG display platform with functional screening for internalizing antibodies using a secondary ADC assay. The discovery strategy entailed immunization of transgenic mice with the cancer antigen ROR2, harboring transgenic IgH and IgL chain gene loci with limited number of fully human V, D, and J gene segments. This was followed by recovering antibody repertoires from the immunized animals, expressing and screening them as full-length human IgG libraries by transposon-mediated display in progenitor B lymphocytes (“Transpo-mAb Display”) for ROR2 binding. Individual cellular “Transpo-mAb” clones isolated by single cell sorting and capable of expressing membrane-bound as well as secreted human IgG were directly screened during antibody discovery, not only for high affinity binding to human ROR2, but also functionally as ADCs using a cytotoxicity assay with a secondary anti-human IgG-toxin-conjugate. Using this strategy, we identified and validated 12 fully human, monoclonal anti-human ROR2 antibodies with nanomolar affinities that are highly potent as ADCs and could be promising candidates for the therapy of human cancer. The screening for functional and internalizing antibodies during the early phase of antibody discovery demonstrates the utility of the mammalian cell-based Transpo-mAb Display platform to select for functional binders and as a powerful tool to improve the efficiency for the development of therapeutically relevant ADCs. |
format | Online Article Text |
id | pubmed-6224377 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-62243772018-11-16 Novel Antibody Drug Conjugates Targeting Tumor-Associated Receptor Tyrosine Kinase ROR2 by Functional Screening of Fully Human Antibody Libraries Using Transpo-mAb Display on Progenitor B Cells Hellmann, Ina Waldmeier, Lorenz Bannwarth-Escher, Marie-Christine Maslova, Kseniya Wolter, Fabian I. Grawunder, Ulf Beerli, Roger R. Front Immunol Immunology Receptor tyrosine kinase-like orphan receptor 2 (ROR2) has been identified as a highly relevant tumor-associated antigen in a variety of cancer indications of high unmet medical need, including renal cell carcinoma and osteosarcoma, making it an attractive target for targeted cancer therapy. Here, we describe the de novo discovery of fully human ROR2-specific antibodies and potent antibody drug conjugates (ADCs) derived thereof by combining antibody discovery from immune libraries of human immunoglobulin transgenic animals using the Transpo-mAb mammalian cell-based IgG display platform with functional screening for internalizing antibodies using a secondary ADC assay. The discovery strategy entailed immunization of transgenic mice with the cancer antigen ROR2, harboring transgenic IgH and IgL chain gene loci with limited number of fully human V, D, and J gene segments. This was followed by recovering antibody repertoires from the immunized animals, expressing and screening them as full-length human IgG libraries by transposon-mediated display in progenitor B lymphocytes (“Transpo-mAb Display”) for ROR2 binding. Individual cellular “Transpo-mAb” clones isolated by single cell sorting and capable of expressing membrane-bound as well as secreted human IgG were directly screened during antibody discovery, not only for high affinity binding to human ROR2, but also functionally as ADCs using a cytotoxicity assay with a secondary anti-human IgG-toxin-conjugate. Using this strategy, we identified and validated 12 fully human, monoclonal anti-human ROR2 antibodies with nanomolar affinities that are highly potent as ADCs and could be promising candidates for the therapy of human cancer. The screening for functional and internalizing antibodies during the early phase of antibody discovery demonstrates the utility of the mammalian cell-based Transpo-mAb Display platform to select for functional binders and as a powerful tool to improve the efficiency for the development of therapeutically relevant ADCs. Frontiers Media S.A. 2018-11-02 /pmc/articles/PMC6224377/ /pubmed/30450096 http://dx.doi.org/10.3389/fimmu.2018.02490 Text en Copyright © 2018 Hellmann, Waldmeier, Bannwarth-Escher, Maslova, Wolter, Grawunder and Beerli. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Immunology Hellmann, Ina Waldmeier, Lorenz Bannwarth-Escher, Marie-Christine Maslova, Kseniya Wolter, Fabian I. Grawunder, Ulf Beerli, Roger R. Novel Antibody Drug Conjugates Targeting Tumor-Associated Receptor Tyrosine Kinase ROR2 by Functional Screening of Fully Human Antibody Libraries Using Transpo-mAb Display on Progenitor B Cells |
title | Novel Antibody Drug Conjugates Targeting Tumor-Associated Receptor Tyrosine Kinase ROR2 by Functional Screening of Fully Human Antibody Libraries Using Transpo-mAb Display on Progenitor B Cells |
title_full | Novel Antibody Drug Conjugates Targeting Tumor-Associated Receptor Tyrosine Kinase ROR2 by Functional Screening of Fully Human Antibody Libraries Using Transpo-mAb Display on Progenitor B Cells |
title_fullStr | Novel Antibody Drug Conjugates Targeting Tumor-Associated Receptor Tyrosine Kinase ROR2 by Functional Screening of Fully Human Antibody Libraries Using Transpo-mAb Display on Progenitor B Cells |
title_full_unstemmed | Novel Antibody Drug Conjugates Targeting Tumor-Associated Receptor Tyrosine Kinase ROR2 by Functional Screening of Fully Human Antibody Libraries Using Transpo-mAb Display on Progenitor B Cells |
title_short | Novel Antibody Drug Conjugates Targeting Tumor-Associated Receptor Tyrosine Kinase ROR2 by Functional Screening of Fully Human Antibody Libraries Using Transpo-mAb Display on Progenitor B Cells |
title_sort | novel antibody drug conjugates targeting tumor-associated receptor tyrosine kinase ror2 by functional screening of fully human antibody libraries using transpo-mab display on progenitor b cells |
topic | Immunology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6224377/ https://www.ncbi.nlm.nih.gov/pubmed/30450096 http://dx.doi.org/10.3389/fimmu.2018.02490 |
work_keys_str_mv | AT hellmannina novelantibodydrugconjugatestargetingtumorassociatedreceptortyrosinekinaseror2byfunctionalscreeningoffullyhumanantibodylibrariesusingtranspomabdisplayonprogenitorbcells AT waldmeierlorenz novelantibodydrugconjugatestargetingtumorassociatedreceptortyrosinekinaseror2byfunctionalscreeningoffullyhumanantibodylibrariesusingtranspomabdisplayonprogenitorbcells AT bannwartheschermariechristine novelantibodydrugconjugatestargetingtumorassociatedreceptortyrosinekinaseror2byfunctionalscreeningoffullyhumanantibodylibrariesusingtranspomabdisplayonprogenitorbcells AT maslovakseniya novelantibodydrugconjugatestargetingtumorassociatedreceptortyrosinekinaseror2byfunctionalscreeningoffullyhumanantibodylibrariesusingtranspomabdisplayonprogenitorbcells AT wolterfabiani novelantibodydrugconjugatestargetingtumorassociatedreceptortyrosinekinaseror2byfunctionalscreeningoffullyhumanantibodylibrariesusingtranspomabdisplayonprogenitorbcells AT grawunderulf novelantibodydrugconjugatestargetingtumorassociatedreceptortyrosinekinaseror2byfunctionalscreeningoffullyhumanantibodylibrariesusingtranspomabdisplayonprogenitorbcells AT beerlirogerr novelantibodydrugconjugatestargetingtumorassociatedreceptortyrosinekinaseror2byfunctionalscreeningoffullyhumanantibodylibrariesusingtranspomabdisplayonprogenitorbcells |