Efficient Methods To Isolate Human Monoclonal Antibodies from Memory B Cells and Plasma Cells

Area of Science:

• Immunology
• Virology
• Vaccine Development

Background:

• Human monoclonal antibodies are crucial for therapeutic and vaccine development.
• Identifying potent, broadly neutralizing antibodies requires efficient screening methods.
• Current methods may not fully capture the diversity of antibody repertoires.

Purpose of the Study:

• To present an optimized method for isolating human monoclonal antibodies from memory B and plasma cells.
• To demonstrate the utility of high-throughput cellular screening for identifying functional antibodies.
• To showcase the application of this approach in discovering novel vaccine targets.

Main Methods:

• Immortalization of memory B cells using Epstein-Barr virus (EBV) and toll-like receptor (TLR) agonists.
• Maintenance of plasma cells in single-cell cultures with interleukin 6 (IL-6) or stromal cells.
• Application of parallel functional assays for high-throughput antibody screening.

Main Results:

• Isolation of potent and broadly neutralizing human monoclonal antibodies against various viruses.
• Identification of a pan-influenza-A-neutralizing antibody and a multi-paramyxovirus-neutralizing antibody.
• Discovery of highly effective cytomegalovirus-neutralizing antibodies targeting a pentameric complex.

Conclusions:

• High-throughput cellular screening of B cell repertoires is instrumental for identifying potent antibodies.
• This target-agnostic approach facilitates the discovery of promising vaccine candidates.
• The identified cytomegalovirus pentameric complex represents a novel vaccine target.