Isolation, cloning and analysis of parvovirus-specific canine antibodies from peripheral blood B cells.

ABSTRACT: B-cell cloning methods enable the analysis of antibody responses against target antigens and can be used to reveal the host antibody repertoire, antigenic sites (epitopes), and details of protective immunity against pathogens. Here, we describe improved methods for isolation of canine peripheral blood B cells producing antibodies against canine parvovirus (CPV) capsids by fluorescence-activated cell sorting, followed by cell cloning. We cultured sorted B cells from an immunized dog in vitro and screened for CPV-specific antibody production. Updated canine-specific primer sets were used to amplify and clone the heavy and light chain immunoglobulin sequences directly from the B cells by reverse transcription and PCR. Monoclonal canine IgGs were produced by cloning heavy and light chain seque

SUBMITTER: Fruh SP 

PROVIDER: S-EPMC10542859 | biostudies-literature | 2023 Oct

REPOSITORIES: biostudies-literature