[Measurement of affinity constant of monoclonal antibody by competitive antibody binding to antigen].
OBJECTIVE
To develop a more reliable and stable method for determining monoclonal antibody (mAb) affinity constant based on competitive antibody/antigen binding.


METHODS
The Kd value was calculated based on the relationship between the binding proportion of the antigen and the original concentration of the mAb that competed for the binding site of the antigen.


RESULTS
The Kd values measured with this improved method under two different conditions were 2.61x10(-12) mol/L and 2.39x10(-12) mol/L, and those with Friguent method in these two conditions were 5.57x10(-10) mol/L and 1.41x10(-10) mol/L, respectively.


CONCLUSIONS
Compared with Friguent method, the Kd values measured with this improved method are closer to the actual value, and the measurement results under different experiment conditions are more stable.
Guo
Jie-biao
JB
Zodolabs Biotechnology Limited Company, Nanchang 330029, China. Efjzlq@tom.com
Guo
Rui
R
Liu
Yan-hua
YH
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology*; D000936:Antigen-Antibody Complex / Q000276:immunology; D001667:Binding, Competitive; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex