دورية أكاديمية

Measuring the sequence-affinity landscape of antibodies with massively parallel titration curves

التفاصيل البيبلوغرافية
العنوان: Measuring the sequence-affinity landscape of antibodies with massively parallel titration curves
المؤلفون: Rhys M Adams, Thierry Mora, Aleksandra M Walczak, Justin B Kinney
المصدر: eLife, Vol 5 (2016)
بيانات النشر: eLife Sciences Publications Ltd, 2016.
سنة النشر: 2016
المجموعة: LCC:Medicine
LCC:Science
LCC:Biology (General)
مصطلحات موضوعية: antibody, affinity, deep mutational scan, titration curve, dissociation constant, Medicine, Science, Biology (General), QH301-705.5
الوصف: 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.
نوع الوثيقة: article
وصف الملف: electronic resource
اللغة: English
تدمد: 2050-084X
Relation: https://elifesciences.org/articles/23156; https://doaj.org/toc/2050-084X
DOI: 10.7554/eLife.23156
URL الوصول: https://doaj.org/article/262b64699e594da586da48dd49c772e1
رقم الأكسشن: edsdoj.262b64699e594da586da48dd49c772e1
قاعدة البيانات: Directory of Open Access Journals
الوصف
تدمد:2050084X
DOI:10.7554/eLife.23156