High-Throughput Monoclonal Antibody Discovery from Phage Libraries: Challenging the Current Preclinical Pipeline to Keep the Pace with the Increasing mAb Demand.

Nicola Zambrano,Marco J Morelli,Margherita Passariello,Alfredo Nicosia,Emanuele Sasso,Guendalina Froechlich,Claudia De Lorenzo,Dejan Lazarevic

doi:10.3390/cancers14051325

Nicola Zambrano, Marco J Morelli + Show 6 more

Open Access

https://doi.org/10.3390/cancers14051325

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Abstract

Simple SummaryMonoclonal antibodies are increasingly used for a broad range of diseases. Rising demand must face with time time-consuming and laborious processes to isolate novel monoclonal antibodies. Next-generation sequencing coupled to phage display provides timely and sustainable high throughput selection strategy to rapidly access novel target. Here, we describe the current NGS-guided strategies to identify potential binders from enriched sub-libraires by applying a user-friendly informatic pipeline to identify and discard false positive clones. Rescue step and strategies to boost mAb yield are also discussed to improve the limiting selection and screening steps.Monoclonal antibodies are among the most powerful therapeutics in modern medicine. Since the approval of the first therapeutic antibody in 1986, monoclonal antibodies keep holding great expectations for application in a range of clinical indications, highlighting the need to provide timely and sustainable access to powerful screening options. However, their application in the past has been limited by time-consuming and expensive steps of discovery and production. The screening of antibody repertoires is a laborious step; however, the implementation of next-generation sequencing-guided screening of single-chain antibody fragments has now largely overcome this issue. This review provides a detailed overview of the current strategies for the identification of monoclonal antibodies from phage display-based libraries. We also discuss the challenges and the possible solutions to improve the limiting selection and screening steps, in order to keep pace with the increasing demand for monoclonal antibodies.

Highlights

In the current era of targeted therapy, monoclonal antibodies represent a powerful therapeutic tool to treat a wide range of diseases
With the approval by the FDA in April 2021 of GlaxoSmithKline’s PD-1/PD-L1 blocker Jemperli, the 100th monoclonal antibody is available for clinical use in the USA
We summarize the next-generation sequencing (NGS)-based technologies applied to phage display and novel approaches to speed up the identification of large numbers of candidate binders, and to improve the yields of recombinant monoclonal antibodies

Summary

Introduction

In the current era of targeted therapy, monoclonal antibodies (mAbs) represent a powerful therapeutic tool to treat a wide range of diseases. The implementation of long accurate reads and the recent introduction of sample multiplexing by barcoding renders the third-generation sequencing platforms very useful to obtain quantitative information of full-length antibody fragments (scFv, Fab) from library screenings. To date, these technologies are still very rarely applied to the screening of antibody libraries, but the recent improvements and a more widespread availability of instruments are paving the way to their more diffused application (Figure 2) [66–68]

Filtering of Potential Binders

Identification of Break Point Selection Cycle

The Issue of False-Positive Clones

Findings

A Computational Strategy for Rapid Discovery of Potential scFv Binders