Real-Time, label-free monitoring of tumor antigen and serum antibody interactions.
Conventional techniques for the detection of biomolecular interactions can be limited by the need for exogenous labels, time- and labor-intensive protocols, as well as by poor sensitivity levels. A refractometer instrument has been reconfigured to detect biomolecular interactions through changes in surface plasmon resonance (SPR). The binding kinetics and affinity values of anti-NY-ESO-1 monoclonal antibody, ES121, to the cancer-testis antigen NY-ESO-1 were determined according to the surface heterogeneity model and resulted in K(D) values of 1.3x10(-9) and 2.1x10(-10) M. The reconfigured instrument was then used to measure the interaction between tumor antigens and serum antibodies against these antigens in preselected cancer patient sera samples. The tumor antigens assayed included NY-ESO-1, SSX2 and p53, all used as recombinant proteins containing polyhistidine tags. These results demonstrated that the instrument is capable of detecting the binding of serum antibodies from cancer patient sera to immobilized tumor antigens, consistent with those observed previously in ELISA-based experiments. These results demonstrate the potential of SPR technology for the rapid diagnosis and monitoring immune responses.
Campagnolo
Christine
C
Field of Microbiology, Cornell University, Ithaca, NY 14853, USA.
Meyers
Katherine J
KJ
Ryan
Thomas
T
Atkinson
Robert C
RC
Chen
Yao-Tseng
YT
Scanlan
Matt J
MJ
Ritter
Gerd
G
Old
Lloyd J
LJ
Batt
Carl A
CA
D000911:Antibodies, Monoclonal / Q000097:blood* / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D000951:Antigens, Neoplasm / Q000097:blood* / Q000276:immunology*; D001774:Blood Chemical Analysis / Q000295:instrumentation / Q000379:methods; D003199:Computer Systems; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods*; D008297:Male; D008565:Membrane Proteins / Q000097:blood* / Q000276:immunology*; D012031:Refractometry / Q000295:instrumentation / Q000379:methods*; D013194:Staining and Labeling; D020349:Surface Plasmon Resonance / Q000379:methods*; D013736:Testicular Neoplasms / Q000097:blood / Q000175:diagnosis / Q000276:immunology
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; C104948:CTAG1B protein, human; D008565:Membrane Proteins