Immunoassays and nucleic acid detection with a biosensor based on surface plasmon resonance.
A technique based on surface plasmon resonance is described which can be used to detect changes of refractive index that occur when one partner of a molecular binding pair diffuses from solution to bind the other partner which is immobilised on a silver surface. Results for the molecular binding pairs; protein-antibody, hapten-antibody and DNA-DNA are described. Instrumentation necessary for implementation of the technique is detailed. Immunoassay of proteins and haptens is possible in less than one minute with a sensitivity of 10(-9) mol/l. Hybridisation of 10 fmoles of a 97 base target sequence on the 1 mm2 area of detection to an immobilised oligonucleotide probe can be detected in less than five minutes. Advantages of the technique include the ability to record the kinetics of binding reactions in "real time" and the lack of labels in this simple assay format. Methods of improving the sensitivity are discussed.
Pollard-Knight
D
D
Amersham International, Pollards Wood Laboratories, Bucks, UK.
Hawkins
E
E
Yeung
D
D
Pashby
D P
DP
Simpson
M
M
McDougall
A
A
Buckle
P
P
Charles
S A
SA
D000818:Animals; D015374:Biosensing Techniques; D004247:DNA / Q000032:analysis*; D006241:Haptens / Q000032:analysis; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods*; D051379:Mice; D009693:Nucleic Acid Hybridization; D011506:Proteins / Q000032:analysis; D012834:Silver
D006241:Haptens; D011506:Proteins; D012834:Silver; D004247:DNA