Bioluminescent Antibodies for Point-of-Care Diagnostics
#MMPMID28510347
Xue L
; Yu Q
; Griss R
; Schena A
; Johnsson K
Angew Chem Int Ed Engl
2017[Jun]; 56
(25
): 7112-7116
PMID28510347
show ga
We introduce a general method to transform antibodies into ratiometric,
bioluminescent sensor proteins for the no-wash quantification of analytes. Our
approach is based on the genetic fusion of antibody fragments to NanoLuc
luciferase and SNAP-tag, the latter being labeled with a synthetic fluorescent
competitor of the antigen. Binding of the antigen, here synthetic drugs, by the
sensor displaces the tethered fluorescent competitor from the antibody and
disrupts bioluminescent resonance energy transfer (BRET) between the luciferase
and fluorophore. The semisynthetic sensors display a tunable response range
(submicromolar to submillimolar) and large dynamic range (?R(max) >500?%), and
they permit the quantification of analytes through spotting of the samples onto
paper followed by analysis with a digital camera.
free
free
free
