A general approach for receptor and antibody-targeted detection of native proteins utilizing split-luciferase reassembly.

ABSTRACT: The direct detection of native proteins in heterogeneous solutions remains a challenging problem. Standard methodologies rely on a separation step to circumvent nonspecific signal generation. We hypothesized that a simple and general method for the detection of native proteins in solution could be achieved through ternary complexation, where the conditional signal generation afforded by split-protein reporters could be married to the specificity afforded by either native receptors or specific antibodies. Toward this goal, we describe a solution phase split-luciferase assay for native protein detection, where we fused fragmented halves of firefly luciferase to separate receptor fragments or single-chain antibodies, allowing for conditional luciferase complementation in the presence of sever

SUBMITTER: Stains CI 

PROVIDER: S-EPMC2955838 | biostudies-literature | 2010 Oct

REPOSITORIES: biostudies-literature