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...
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doaj-a9c44bbc4a8840b5941f96c57d32a5bc2020-11-25T02:32:27ZengMDPI AGAntibodies2073-44682019-08-01834510.3390/antib8030045antib8030045Back-to-Germline (B2G) Procedure for Antibody DevolutionAnja Schrade0Alexander Bujotzek1Christian Spick2Martina Wagner3Johannes Goerl4Xenia Wezler5Guy Georges6Roland E. Kontermann7Ulrich Brinkmann8Roche Pharma Research and Early Development (pRED), Large Molecule Research (LMR), Roche Innovation Center Munich, 82377 Penzberg, GermanyRoche Pharma Research and Early Development (pRED), Large Molecule Research (LMR), Roche Innovation Center Munich, 82377 Penzberg, GermanyRoche Pharma Research and Early Development (pRED), Large Molecule Research (LMR), Roche Innovation Center Munich, 82377 Penzberg, GermanyRoche Pharma Research and Early Development (pRED), Large Molecule Research (LMR), Roche Innovation Center Munich, 82377 Penzberg, GermanyRoche Pharma Research and Early Development (pRED), Large Molecule Research (LMR), Roche Innovation Center Munich, 82377 Penzberg, GermanyRoche Pharma Research and Early Development (pRED), Large Molecule Research (LMR), Roche Innovation Center Munich, 82377 Penzberg, GermanyRoche Pharma Research and Early Development (pRED), Large Molecule Research (LMR), Roche Innovation Center Munich, 82377 Penzberg, GermanyInstitute of Cell Biology & Immunology, Stuttgart University, 70569 Stuttgart, GermanyRoche Pharma Research and Early Development (pRED), Large Molecule Research (LMR), Roche Innovation Center Munich, 82377 Penzberg, GermanyBispecific 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.https://www.mdpi.com/2073-4468/8/3/45protein engineeringantibodymaturationaffinitystructureantigen binding |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Anja Schrade Alexander Bujotzek Christian Spick Martina Wagner Johannes Goerl Xenia Wezler Guy Georges Roland E. Kontermann Ulrich Brinkmann |
spellingShingle |
Anja Schrade Alexander Bujotzek Christian Spick Martina Wagner Johannes Goerl Xenia Wezler Guy Georges Roland E. Kontermann Ulrich Brinkmann Back-to-Germline (B2G) Procedure for Antibody Devolution Antibodies protein engineering antibody maturation affinity structure antigen binding |
author_facet |
Anja Schrade Alexander Bujotzek Christian Spick Martina Wagner Johannes Goerl Xenia Wezler Guy Georges Roland E. Kontermann Ulrich Brinkmann |
author_sort |
Anja Schrade |
title |
Back-to-Germline (B2G) Procedure for Antibody Devolution |
title_short |
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_sort |
back-to-germline (b2g) procedure for antibody devolution |
publisher |
MDPI AG |
series |
Antibodies |
issn |
2073-4468 |
publishDate |
2019-08-01 |
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. |
topic |
protein engineering antibody maturation affinity structure antigen binding |
url |
https://www.mdpi.com/2073-4468/8/3/45 |
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