High-throughput strategies for monoclonal antibody screening: advances and challenges

Abstract

Antibodies characterized by high affinity and specificity, developed through high-throughput screening and rapid preparation, are crucial to contemporary biomedical industry. Traditional antibody preparation via the hybridoma strategy faces challenges like low efficiency, long manufacturing cycles, batch variability and labor intensity. Advances in molecular biology and gene editing technologies offer revolutionary improvements in antibody production. New high-throughput technologies like antibody library display, single B cell antibody technologies, and single-cell sequencing have significantly cut costs and boosted the efficiency of antibody development. These innovations accelerate commercial applications of antibodies, meeting the biopharmaceutical industry's evolving demands. This review explores recent advancements in high-throughput development of antibody, highlighting their potential advantages over traditional methods and their promising future.

Keywords: Antibody preparation; Display system for antibody libraries; High-throughput; Single B cell antibody; Single-cell sequencing.

Fig. 1

The high-throughput antibody production scheme of the antibody library display technology. Antibody libraries from immunized animals and humans are displayed on a variety of vectors (e.g. phage, yeast, ribosomes, etc.) to construct diverse antibody libraries, and high-affinity binding vectors are isolated through multiple rounds of iterative screening. The gene sequences of the variable regions of the antibodies were determined by ELISA identification and sequencing analysis. The gene sequences of the variable regions are integrated into the designated expression vectors for antibody production, followed by downstream characterization and analysis

Fig. 2

The high-throughput antibody production scheme of the single B cell antibody technology. In the context of isolating PBMCs or splenocytes from immunized animals, as well as PBMCs from convalescent patients, various techniques such as MACS can be employed to enhance the concentration of target B cells. Subsequently, antigen-specific single B cells be isolated using methods such as FACS and microfluidics. The antibody variable region genes can be cloned through RT-PCR to facilitate antibody expression and subsequent analysis

Fig. 3

The high-throughput antibody production scheme of the single-cell sequencing. In the context of isolating PBMCs or splenocytes from immunized animals, as well as PBMCs from convalescent patients. Target B cells were enriched using MACS or FACS. Single-cell sequencing was conducted on the enriched cells to obtain 5'-end transcriptome data and immune repertoire data. Bioinformatics analysis was then employed to identify high-affinity antibody gene sequences from a vast amount of single-cell sequencing data. Specific vectors were subsequently incorporated for antibody expression and downstream characterization