Development of GFP-based biosensors possessing the binding properties of antibodies.
Proc Natl Acad Sci U S A
; 106(29): 11895-900, 2009 Jul 21.
Article
em En
| MEDLINE
| ID: mdl-19574456
ABSTRACT
Proteins that can bind specifically to targets that also have an intrinsic property allowing for easy detection could facilitate a multitude of applications. While the widely used green fluorescent protein (GFP) allows for easy detection, attempts to insert multiple binding loops into GFP to impart affinity for a specific target have been met with limited success because of the structural sensitivity of the GFP chromophore. In this study, directed evolution using a surrogate loop approach and yeast surface display yielded a family of GFP scaffolds capable of accommodating 2 proximal, randomized binding loops. The library of potential GFP-based binders or ''GFAbs'' was subsequently mined for GFAbs capable of binding to protein targets. Identified GFAbs bound with nanomolar affinity and required binding contributions from both loops indicating the advantage of a dual loop GFAb platform. Finally, GFAbs were solubly produced and used as fluorescence detection reagents to demonstrate their utility.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Técnicas Biossensoriais
/
Proteínas de Fluorescência Verde
/
Anticorpos
Tipo de estudo:
Clinical_trials
Idioma:
En
Ano de publicação:
2009
Tipo de documento:
Article