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Microbiol Spectr. 2014 Oct;2(5). doi: 10.1128/microbiolspec.AID-0018-2014.

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

Author information

1
Humabs BioMed SA, 6500 Bellinzona, Switzerland.
2
Institute for Research in Biomedicine, 6500 Bellinzona, Switzerland.

Abstract

In this article, we highlight the advantages of isolating human monoclonal antibodies from the human memory B cells and plasma cell repertoires by using high-throughput cellular screens. Memory B cells are immortalized with high efficiency using Epstein-Barr virus (EBV) in the presence of a toll-like receptor (TLR) agonist, while plasma cells are maintained in single-cell cultures by using interleukin 6 (IL-6) or stromal cells. In both cases, multiple parallel assays, including functional assays, can be used to identify rare cells that produce antibodies with unique properties. Using these methods, we have isolated potent and broadly neutralizing antibodies against a variety of viruses, in particular, a pan-influenza-A-neutralizing antibody and an antibody that neutralizes four different paramyxoviruses. Given the high throughput and the possibility of directly screening for function (rather than just binding), these methods are instrumental to implement a target-agnostic approach to identify the most effective antibodies and, consequently, the most promising targets for vaccine design. This approach is exemplified by the identification of unusually potent cytomegalovirus-neutralizing antibodies that led to the identification of the target, a pentameric complex that we are developing as a candidate vaccine.

[Indexed for MEDLINE]

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