In:
The Journal of Immunology, The American Association of Immunologists, Vol. 200, No. 1_Supplement ( 2018-05-01), p. 174.2-174.2
Abstract:
Next-Generation sequencing has become an essential tool in the analysis of antibody responses in the settings of health, vaccination, and disease. However, immune receptors comprise two chains encoded by separate mRNA strands, and conventional NextGen sequencing fails to identify the native pairings encoded by individual lymphocytes. To overcome this limitation we have applied recent technical advances in high-throughput sequencing and functional analysis of complete antibodies (i.e., paired heavy and light chain sequencing) to generate a comprehensive understanding of the antibody response to vaccination and natural infection. Here we present a new technology to screen natively-paired human antibody repertoires from millions of B cells. Libraries of natively-paired variable region heavy and light (VH:VL) amplicons were expressed in a yeast display platform that was optimized for human Fab surface expression, and the resulting libraries were interrogated for binding to viral vaccine antigens via FACS paired with next generation sequencing. Using our method we identified HIV-1 broadly neutralizing antibodies (bNAbs) from an HIV-1 slow progressor and high-affinity neutralizing antibodies responding to an Ebola virus glycoprotein vaccination. These next-generation approaches are providing detailed molecular feedback on immune receptor responses and are informing the design and discovery of new vaccines and therapeutics.
Type of Medium:
Online Resource
ISSN:
0022-1767
,
1550-6606
DOI:
10.4049/jimmunol.200.Supp.174.2
Language:
English
Publisher:
The American Association of Immunologists
Publication Date:
2018
detail.hit.zdb_id:
1475085-5
