A robust high throughput platform to generate functional recombinant monoclonal antibodies using rabbit B cells from peripheral blood.

ABSTRACT: We have developed a robust platform to generate and functionally characterize rabbit-derived antibodies using B cells from peripheral blood. The rapid high throughput procedure generates a diverse set of antibodies, yet requires only few animals to be immunized without the need to sacrifice them. The workflow includes (i) the identification and isolation of single B cells from rabbit blood expressing IgG antibodies, (ii) an elaborate short term B-cell cultivation to produce sufficient monoclonal antigen specific IgG for comprehensive phenotype screens, (iii) the isolation of VH and VL coding regions via PCR from B-cell clones producing antigen specific and functional antibodies followed by the sequence determination, and (iv) the recombinant expression and purification of IgG antibodies. T

SUBMITTER: Seeber S 

PROVIDER: S-EPMC3913575 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature