title: FACS binding assay for analysing GDNF interactions.
abstract: Glial cell-line derived neurotrophic factor (GDNF) is a secreted protein with great therapeutic potential. However, in order to analyse the interactions between GDNF and its receptors, researchers have been mostly dependent of radioactive binding assays. We developed a FACS-based binding assay for GDNF as an alternative to current methods. We demonstrated that the FACS-based assay using TGW cells allowed readily detection of GDNF binding and displacement to endogenous receptors. The dissociation constant and half maximal inhibitory concentration obtained were comparable to other studies using standard binding assays. Overall, this FACS-based, simple to perform and adaptable to high throughput setup, provides a safer and reliable alternative to radioactive methods.
authors.affiliation: CNS Gene Therapy, Department of Experimental Medical Sciences, Faculty of Medicine, Lund University, Lund, Sweden.
authors.forename: Luís
authors.initials: L
authors.lastname: Quintino
authors.forename: Aurélie
authors.initials: A
authors.lastname: Baudet
authors.forename: Jonas
authors.initials: J
authors.lastname: Larsson
authors.forename: Cecilia
authors.initials: C
authors.lastname: Lundberg
mesh_terms: D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods*; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000032:analysis* / Q000378:metabolism*; D006801:Humans; D009474:Neurons / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology
chemical_list: D051100:Glial Cell Line-Derived Neurotrophic Factor
keywords: 293T cells expressing GFRα1; 293T-GFR1; Binding assay; DOL; Degree of labelling; FACS; Fluorescent-activated cell sorting; G-CSF; GDNF; GDNF Family receptor alpha; GDNF conjugated with Alexa 488; GDNF(488); GFRα; Glial cell-line derived neurotrophic factor; granulocyte-colony stimulating factor