Beyond Single Clones: High-Throughput Sequencing in Antibody Discovery

Ahmed S Fahad; Matías F Gutiérrez-Gonzalez; Bharat Madan; Brandon J DeKosky

doi:10.1101/pdb.top107772

Beyond Single Clones: High-Throughput Sequencing in Antibody Discovery

Cold Spring Harb Protoc. 2025 Jan 2;2025(1):pdb.top107772. doi: 10.1101/pdb.top107772.

Authors

Ahmed S Fahad^#¹, Matías F Gutiérrez-Gonzalez^#¹, Bharat Madan¹, Brandon J DeKosky^{2

3}

Affiliations

¹ The Ragon Institute of Massachusetts General Hospital, Massachusetts Institute of Technology and Harvard University, Cambridge, Massachusetts 02139, USA.
² The Ragon Institute of Massachusetts General Hospital, Massachusetts Institute of Technology and Harvard University, Cambridge, Massachusetts 02139, USA dekosky@mit.edu.
³ Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.

^# Contributed equally.

PMID: 39586681
DOI: 10.1101/pdb.top107772

Abstract

Antibody repertoire sequencing and display library screening are powerful approaches for antibody discovery and engineering that can connect DNA sequence with antibody function. Antibody display and screening studies have made a tremendous impact on immunology and biotechnology over the last decade, accelerated by technological advances in high-throughput DNA sequencing techniques. Indeed, bioinformatic analysis of antibody DNA library data has now taken a central role in modern antibody drug discovery, and is also critical for many ongoing studies of human immune development. Here, we describe current trends in antibody DNA library screening and analysis, and introduce a selection of protocols describing fundamental bioinformatic techniques to enable scientists to efficiently study antibody DNA libraries.

MeSH terms

Antibodies* / genetics
Computational Biology / methods
Gene Library*
High-Throughput Nucleotide Sequencing* / methods
Humans

Substances

Antibodies