Antibody therapies for the prevention and treatment of viral infections

Salazar, Georgina; Zhang, Ningyan; Fu, Tong-Ming; An, Zhiqiang

Deep sequencing of paired antibodies encoding genes from B cells. While cloning mAbs from individual memory or plasma B cells is a robust and efficient approach to isolate viral-specific antibodies, the approach provides only a snapshot and sample of the antibody genetic repertoire of an individual at a given time against a particular virus. Various deep-sequencing approaches can provide an unbiased picture of the B-cell repertoire based on either its heavy chain or light chain information, but the endogenous pairing of heavy and light chain is lost after bulk lysis of B-cell populations. 40 Phylogenetic matching of heavy and light chains from antibody deep-sequencing data can provide an approximation of natural heavy and light chain pairing. 41 Highthroughput sequencing of paired heavy and light chain variable Fig. 3 Overview of isolation, culture, and antibody gene cloning from plasma B cells. This method can be an efficient way of generating antibodies with desired specificity given the right antigen bait and timing. The diagram was generated based on a combination of publications 15, 16, [28] [29] [30] [31] [32] [33] [34] [35] [36] [37] regions at the single-cell level has proven technically challenging. A method that allows the determination of a paired heavy and light chain variable region repertoire from millions of cells with high precision was recently reported. 19, 20, 42 Briefly, flow-focusing is used to encapsulate single cells in emulsions containing magnetic beads for messenger RNA (mRNA) capture. The mRNA transcripts are then reverse-transcribed, physically linked to their partners by overlap extension PCR, and interrogated by highthroughput paired-end sequencing. 19 Once adapted widely by the research community, this method will provide powerful information on antibody response to infection and vaccination, vaccine efficacy, and the cloning of potent viral neutralizing antibodies. 42 mAbs targeting viral infections Table 1 shows mAbs in clinical development for the prevention and treatment of infectious viral diseases. These mAbs have been isolated from a variety of non-immune and immune sources, human and animal donors, using the range of strategies for generation of therapeutic antibodies introduced above.