Single-Cell Droplet Microfluidic Screening for Antibodies Specifically Binding to Target Cells

Cell Reports
Nachiket ShembekarChristoph A Merten

Abstract

Monoclonal antibodies are a main player in modern drug discovery. Many antibody screening formats exist, each with specific advantages and limitations. Nonetheless, it remains challenging to screen antibodies for the binding of cell-surface receptors (the most important class of all drug targets) or for the binding to target cells rather than purified proteins. Here, we present a high-throughput droplet microfluidics approach employing dual-color normalized fluorescence readout to detect antibody binding. This enables us to obtain quantitative data on target cell recognition, using as little as 33 fg of IgG per assay. Starting with an excess of hybridoma cells releasing unspecific antibodies, individual clones secreting specific binders (of target cells co-encapsulated into droplets) could be enriched 220-fold after sorting 80,000 clones in a single experiment. This opens the way for therapeutic antibody discovery, especially since the single-cell approach is in principle also applicable to primary human plasma cells.

Citations

Sep 15, 2018·Nanotechnology·Ziyi HePeng Li
Nov 7, 2018·Lab on a Chip·Aude I SegalinyWeian Zhao
Apr 19, 2019·Angewandte Chemie·Xuewei WangMark E Meyerhoff
Jul 17, 2020·The Journal of Immunology : Official Journal of the American Association of Immunologists·Klaus EyerJean Baudry
Aug 12, 2020·BioTechniques·Jane Ru Choi
Feb 16, 2019·Proceedings of the National Academy of Sciences of the United States of America·Venkata YelleswarapuDavid Issadore
Nov 22, 2018·Frontiers in Immunology·Nidhi SinhaJurjen Tel
May 28, 2020·Scientific Reports·Muhsincan Sesen, Graeme Whyte
Nov 9, 2019·Microsystems & Nanoengineering·Lucas ArmbrechtPetra Stephanie Dittrich
Jun 22, 2018·High-throughput·Clément RegnaultAxel Hochstetter
Aug 14, 2020·Nature Protocols·Yacine BounabChristophe Védrine
Dec 10, 2020·Lab on a Chip·Renchang ZhangLin Gui
Dec 5, 2020·Journal of Controlled Release : Official Journal of the Controlled Release Society·Samuel K LaiRaymond J Pickles
Jan 29, 2021·ACS Applied Materials & Interfaces·Wenshuai WuYing Mu
Jan 3, 2021·Drug Resistance Updates : Reviews and Commentaries in Antimicrobial and Anticancer Chemotherapy·Chuan YangLiwu Fu
Jan 14, 2020·Current Opinion in Biotechnology·Marion ArnaudAlexandre Harari
Jan 1, 2021·Biomicrofluidics·Michael S GerltJürg Dual
May 23, 2020·Acta Biomaterialia·Kaitlyn BaconStefano Menegatti
Mar 19, 2021·Critical Reviews in Biotechnology·Jianhua YangLeilei Zhu
Mar 25, 2021·Biological Chemistry·Bernhard ValldorfStefan Zielonka
Mar 19, 2021·European Journal of Immunology·Olivia T M BucheliKlaus Eyer
Aug 1, 2021·Drug Discovery Today·Boqian WangXianting Ding
Dec 7, 2019·Analytical Chemistry·Malgorzata A WitekSteven A Soper
Sep 11, 2021·Lab on a Chip·Julie Van LentJeroen Lammertyn
Sep 30, 2021·MAbs·Ramona GaaAchim Doerner
Oct 8, 2021·Lab on a Chip·Saurabh VyawahareAndrew Homyk

❮ Previous
Next ❯

Methods Mentioned

BETA
Assay
chip
PCR
Fluorescence
flow cytometry

Software Mentioned

LabVIEW

Related Concepts

Related Feeds