Measuring the sequence-affinity landscape of antibodies with massively parallel titration curves
Despite the central role that antibodies play in the adaptive immune system and in biotechnology, much remains unknown about the quantitative relationship between an antibody’s amino acid sequence and its antigen binding affinity. Here we describe a new experimental approach, called Tite-Seq, that i...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2016-12-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/23156 |
| _version_ | 1818018832877355008 |
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| author | Rhys M Adams Thierry Mora Aleksandra M Walczak Justin B Kinney |
| author_facet | Rhys M Adams Thierry Mora Aleksandra M Walczak Justin B Kinney |
| author_sort | Rhys M Adams |
| collection | DOAJ |
| description | Despite the central role that antibodies play in the adaptive immune system and in biotechnology, much remains unknown about the quantitative relationship between an antibody’s amino acid sequence and its antigen binding affinity. Here we describe a new experimental approach, called Tite-Seq, that is capable of measuring binding titration curves and corresponding affinities for thousands of variant antibodies in parallel. The measurement of titration curves eliminates the confounding effects of antibody expression and stability that arise in standard deep mutational scanning assays. We demonstrate Tite-Seq on the CDR1H and CDR3H regions of a well-studied scFv antibody. Our data shed light on the structural basis for antigen binding affinity and suggests a role for secondary CDR loops in establishing antibody stability. Tite-Seq fills a large gap in the ability to measure critical aspects of the adaptive immune system, and can be readily used for studying sequence-affinity landscapes in other protein systems. |
| first_indexed | 2024-04-14T07:45:01Z |
| format | Article |
| id | doaj.art-262b64699e594da586da48dd49c772e1 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-14T07:45:01Z |
| publishDate | 2016-12-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-262b64699e594da586da48dd49c772e12022-12-22T02:05:22ZengeLife Sciences Publications LtdeLife2050-084X2016-12-01510.7554/eLife.23156Measuring the sequence-affinity landscape of antibodies with massively parallel titration curvesRhys M Adams0Thierry Mora1https://orcid.org/0000-0002-5456-9361Aleksandra M Walczak2https://orcid.org/0000-0002-2686-5702Justin B Kinney3https://orcid.org/0000-0003-1897-3778Laboratoire de Physique Théorique, UMR8549, CNRS, École Normale Supérieure, Paris, France; Simons Center for Quantitative Biology, Cold Spring Harbor Laboratory, Cold Spring Harbor, United StatesLaboratoire de Physique Statistique, UMR8550, CNRS, École Normale Supérieure, Paris, FranceLaboratoire de Physique Théorique, UMR8549, CNRS, École Normale Supérieure, Paris, FranceSimons Center for Quantitative Biology, Cold Spring Harbor Laboratory, Cold Spring Harbor, United StatesDespite the central role that antibodies play in the adaptive immune system and in biotechnology, much remains unknown about the quantitative relationship between an antibody’s amino acid sequence and its antigen binding affinity. Here we describe a new experimental approach, called Tite-Seq, that is capable of measuring binding titration curves and corresponding affinities for thousands of variant antibodies in parallel. The measurement of titration curves eliminates the confounding effects of antibody expression and stability that arise in standard deep mutational scanning assays. We demonstrate Tite-Seq on the CDR1H and CDR3H regions of a well-studied scFv antibody. Our data shed light on the structural basis for antigen binding affinity and suggests a role for secondary CDR loops in establishing antibody stability. Tite-Seq fills a large gap in the ability to measure critical aspects of the adaptive immune system, and can be readily used for studying sequence-affinity landscapes in other protein systems.https://elifesciences.org/articles/23156antibodyaffinitydeep mutational scantitration curvedissociation constant |
| spellingShingle | Rhys M Adams Thierry Mora Aleksandra M Walczak Justin B Kinney Measuring the sequence-affinity landscape of antibodies with massively parallel titration curves eLife antibody affinity deep mutational scan titration curve dissociation constant |
| title | Measuring the sequence-affinity landscape of antibodies with massively parallel titration curves |
| title_full | Measuring the sequence-affinity landscape of antibodies with massively parallel titration curves |
| title_fullStr | Measuring the sequence-affinity landscape of antibodies with massively parallel titration curves |
| title_full_unstemmed | Measuring the sequence-affinity landscape of antibodies with massively parallel titration curves |
| title_short | Measuring the sequence-affinity landscape of antibodies with massively parallel titration curves |
| title_sort | measuring the sequence affinity landscape of antibodies with massively parallel titration curves |
| topic | antibody affinity deep mutational scan titration curve dissociation constant |
| url | https://elifesciences.org/articles/23156 |
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