Determination of affinities and antigenic epitopes of bovine cardiac troponin I (cTnI) with monoclonal antibodies by surface plasmon resonance biosensor.
A surface plasmon resonance (SPR) biosensor based on wavelength modulation was used for real-time detection of the interaction of three monoclonal antibodies and antigens of bovine cardiac troponin I (cTnI). In order to recognize antigenic epitopes of bovine cTnI, two experimental modes were applied. In the first experimental mode, three monoclonal antibodies were divided into three groups and three experiments were performed on biosensor surfaces prepared with protein A. In the second experimental mode, antigen was immobilized on the biosensor surface prepared by the amine-coupling method and three monoclonal antibodies were detected in turn. The results obtained by the two modes are consistent. In addition, the affinities of the monoclonal antibodies for the antigen were also determined by the association rate and the disassociation rate in real-time. These results validate the biosensor technology and illustrate how biosensors based on wavelength modulation can be used to study the interaction of monoclonal antibodies and antigens in real time.
Liu
Xia
X
College of Chemistry, Jilin University, Changchun 130023, People's Republic of China.
Wei
Jingyan
J
Song
Daqian
D
Zhang
Ziwei
Z
Zhang
Hanqi
H
Luo
Guimin
G
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001666:Binding Sites, Antibody / Q000276:immunology; D015374:Biosensing Techniques / Q000379:methods*; D002417:Cattle; D000939:Epitopes / Q000032:analysis* / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000737:chemistry / Q000276:immunology; D007700:Kinetics; D018448:Models, Immunological; D008970:Molecular Weight; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000276:immunology; D020349:Surface Plasmon Resonance / Q000379:methods*; D013997:Time Factors; D019210:Troponin I / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007128:Immunoglobulin Fragments; D013205:Staphylococcal Protein A; D019210:Troponin I