Isolation of single HIV-1 Envelope specific B cells and antibody cloning from immunized rhesus macaques.

ABSTRACT: Antibody cloning from single B cells is an essential tool for characterizing humoral immune responses and obtaining valuable therapeutic and analytical reagents. Antibody cloning from individuals with high serologic titers to HIV-1, Influenza, Malaria and ZIKV has led to new insights that inform vaccine design efforts. In contrast to humans and mice, less is known about antibody cloning from single B cells in macaques. Here, we describe a protocol to identify and purify single antigen-specific macaque B cells, and subsequently clone and produce macaque monoclonal antibodies. The sorting strategy requires the use of a combination of fluorochrome labeled antigens and omission of anti-IgG antibodies that can interfere with antigen binding and vice versa. Optimized methods for macaque antibody

SUBMITTER: Wang Z 

PROVIDER: S-EPMC6961706 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature