Objective: To measure scFv antibody af finity using non-competitive enzyme-linked immunosorbent assay (ELISA). Methods:Using serial dilutions of antigen (, ) and antibody (anti-MAGE-A1 scFv antibody), the affinity co...Objective: To measure scFv antibody af finity using non-competitive enzyme-linked immunosorbent assay (ELISA). Methods:Using serial dilutions of antigen (, ) and antibody (anti-MAGE-A1 scFv antibody), the affinity constant (K aff) was measured by non-competitive ELISA. Two sigmoid curves of optical density (OD) v ersus logarithms of antibody concentrations were estimated by SPSS 10.0 software . The maximum value of OD (OD-100) of each curve was computed respectively and OD-50, half of OD-100, was obtained. Then the concentrations of antibody corre sponding to OD-50 on the curves, named t and t were calculated . For =1/2 , K aff=1/{2 t- t}. Results: The affinity constant of scFv antibody was about 1.432×1 06 L/mol. Conclusion: Based on the Law of Mass Action, n on-competitive ELISA method for measurement of antibody-antigen affinity const ant is simple, rapid and reliable.展开更多
基金Science Fund of Department of Health of Jiangsu Province(H200103)
文摘Objective: To measure scFv antibody af finity using non-competitive enzyme-linked immunosorbent assay (ELISA). Methods:Using serial dilutions of antigen (, ) and antibody (anti-MAGE-A1 scFv antibody), the affinity constant (K aff) was measured by non-competitive ELISA. Two sigmoid curves of optical density (OD) v ersus logarithms of antibody concentrations were estimated by SPSS 10.0 software . The maximum value of OD (OD-100) of each curve was computed respectively and OD-50, half of OD-100, was obtained. Then the concentrations of antibody corre sponding to OD-50 on the curves, named t and t were calculated . For =1/2 , K aff=1/{2 t- t}. Results: The affinity constant of scFv antibody was about 1.432×1 06 L/mol. Conclusion: Based on the Law of Mass Action, n on-competitive ELISA method for measurement of antibody-antigen affinity const ant is simple, rapid and reliable.
