Affinity determination of biotinylated antibodies by flow cytometry.
Affinity determination is a crucial step of an antibody characterization. Here, we describe a method for antibody affinity determination by flow cytometry, relying on the unique affinity of biotin for streptavidin for easy and efficient antibody labeling. Several labeling approaches are described and discussed in this chapter, including chemical and enzymatic (in vivo and in vitro) biotinylation. Finally, a procedure for K                 (D) determination by flow cytometry is precisely described.
INSERM U624, Marseille, France.
Klervi
K
Even-Desrumeaux
Patrick
P
Chames
D000906:Antibodies / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology*; D019921:Biotinylation; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007118:Immunoassay; D012044:Regression Analysis
D000906:Antibodies