Cargando…

Back-to-Germline (B2G) Procedure for Antibody Devolution

Bispecific antibodies (bsAbs) with avidity-enhanced specificity can be used to address target cells with increased specificity, ideally binding efficiently to cells that express two cognate antigens, yet not to cells that express only one of those. Building blocks required to generate such bsAbs are...

Descripción completa

Detalles Bibliográficos
Autores principales: Schrade, Anja, Bujotzek, Alexander, Spick, Christian, Wagner, Martina, Goerl, Johannes, Wezler, Xenia, Georges, Guy, Kontermann, Roland E., Brinkmann, Ulrich
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6784197/
https://www.ncbi.nlm.nih.gov/pubmed/31544851
http://dx.doi.org/10.3390/antib8030045
_version_ 1783457714390695936
author Schrade, Anja
Bujotzek, Alexander
Spick, Christian
Wagner, Martina
Goerl, Johannes
Wezler, Xenia
Georges, Guy
Kontermann, Roland E.
Brinkmann, Ulrich
author_facet Schrade, Anja
Bujotzek, Alexander
Spick, Christian
Wagner, Martina
Goerl, Johannes
Wezler, Xenia
Georges, Guy
Kontermann, Roland E.
Brinkmann, Ulrich
author_sort Schrade, Anja
collection PubMed
description Bispecific antibodies (bsAbs) with avidity-enhanced specificity can be used to address target cells with increased specificity, ideally binding efficiently to cells that express two cognate antigens, yet not to cells that express only one of those. Building blocks required to generate such bsAbs are binders that recognize the two antigens with high specificity yet with various (including very low monovalent) affinities. The herein described ‘back-to-germline’ (B2G) procedure defines such derivatives. It converts parent antibodies with high specificity to derivatives that retain specificity but modulate affinity. The approach defines mutations to be introduced into antibody complementarity-determining regions (CDRs) regions without requiring structures of antibody-antigen complexes. Instead, it reverses the B-cell maturation process that increases affinities, with preference on CDR residues with high antigen contact probability. Placing germline residues at those positions generates VH and VL domains and Fv-combinations thereof that retain specificities but are ‘de-matured’ to different degrees. De-maturation influences on-rates and off-rates, and can produce entities with extremely low affinity for which binding can only be detected in bivalent formats. A comparison with alanine replacement in CDRs (so far, the most frequently applied technology) indicates that B2G may be more reliable/predictable without introduction of stickiness or poly-reactivity. The applicability for generating sets of affinity-modulated monospecific variants is exemplarily shown for antibodies that bind CD138, Her2/neu, and EGFR.
format Online
Article
Text
id pubmed-6784197
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-67841972019-10-16 Back-to-Germline (B2G) Procedure for Antibody Devolution Schrade, Anja Bujotzek, Alexander Spick, Christian Wagner, Martina Goerl, Johannes Wezler, Xenia Georges, Guy Kontermann, Roland E. Brinkmann, Ulrich Antibodies (Basel) Article Bispecific antibodies (bsAbs) with avidity-enhanced specificity can be used to address target cells with increased specificity, ideally binding efficiently to cells that express two cognate antigens, yet not to cells that express only one of those. Building blocks required to generate such bsAbs are binders that recognize the two antigens with high specificity yet with various (including very low monovalent) affinities. The herein described ‘back-to-germline’ (B2G) procedure defines such derivatives. It converts parent antibodies with high specificity to derivatives that retain specificity but modulate affinity. The approach defines mutations to be introduced into antibody complementarity-determining regions (CDRs) regions without requiring structures of antibody-antigen complexes. Instead, it reverses the B-cell maturation process that increases affinities, with preference on CDR residues with high antigen contact probability. Placing germline residues at those positions generates VH and VL domains and Fv-combinations thereof that retain specificities but are ‘de-matured’ to different degrees. De-maturation influences on-rates and off-rates, and can produce entities with extremely low affinity for which binding can only be detected in bivalent formats. A comparison with alanine replacement in CDRs (so far, the most frequently applied technology) indicates that B2G may be more reliable/predictable without introduction of stickiness or poly-reactivity. The applicability for generating sets of affinity-modulated monospecific variants is exemplarily shown for antibodies that bind CD138, Her2/neu, and EGFR. MDPI 2019-08-26 /pmc/articles/PMC6784197/ /pubmed/31544851 http://dx.doi.org/10.3390/antib8030045 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Schrade, Anja
Bujotzek, Alexander
Spick, Christian
Wagner, Martina
Goerl, Johannes
Wezler, Xenia
Georges, Guy
Kontermann, Roland E.
Brinkmann, Ulrich
Back-to-Germline (B2G) Procedure for Antibody Devolution
title Back-to-Germline (B2G) Procedure for Antibody Devolution
title_full Back-to-Germline (B2G) Procedure for Antibody Devolution
title_fullStr Back-to-Germline (B2G) Procedure for Antibody Devolution
title_full_unstemmed Back-to-Germline (B2G) Procedure for Antibody Devolution
title_short Back-to-Germline (B2G) Procedure for Antibody Devolution
title_sort back-to-germline (b2g) procedure for antibody devolution
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6784197/
https://www.ncbi.nlm.nih.gov/pubmed/31544851
http://dx.doi.org/10.3390/antib8030045
work_keys_str_mv AT schradeanja backtogermlineb2gprocedureforantibodydevolution
AT bujotzekalexander backtogermlineb2gprocedureforantibodydevolution
AT spickchristian backtogermlineb2gprocedureforantibodydevolution
AT wagnermartina backtogermlineb2gprocedureforantibodydevolution
AT goerljohannes backtogermlineb2gprocedureforantibodydevolution
AT wezlerxenia backtogermlineb2gprocedureforantibodydevolution
AT georgesguy backtogermlineb2gprocedureforantibodydevolution
AT kontermannrolande backtogermlineb2gprocedureforantibodydevolution
AT brinkmannulrich backtogermlineb2gprocedureforantibodydevolution