Investigation of the feasibility of SPR as a method for detection of Troponin-I by antibody binding for whole blood analysis.
Author
Poulsen, Morten
Term
4. term
Education
Publication year
2010
Submitted on
2010-06-30
Pages
32
Abstract
Dette projekt giver en introduktion til Surface Plasmon Resonans (SPR), en optisk metode, der kan måle, når molekyler binder sig til en sensoroverflade ved at registrere ændringer i reflekteret lys. Der er udviklet et Matlab-program til en teoretisk model af en flerlaget SPR-overflade, som bruges til at vurdere, hvor godt antistoffer kan fastgøres (immobiliseres) på overfladen. Projektet beskriver også en måde at fremstille SPR-overflader ud fra en poleret glaswafer samt en procedure til at immobilisere antistoffer på disse overflader. Denne procedure anvendes til at bygge en biosensor mod hjertemarkøren Troponin I (cTnI), og sensoren testes. En kommercielt tilgængelig SPR-chip (Xantec SPR sensor chip CMD500l) testes også og giver mere reproducerbare data. Når sensoren testes på cTnI fortyndet i bovint serumalbumin, er resultaterne dog svage. Der konkluderes, at der er behov for flere forsøg med forskellige pH-værdier og forskellige overfladebelægninger.
This project introduces Surface Plasmon Resonance (SPR), an optical technique that detects when molecules bind to a sensor surface by measuring changes in reflected light. A Matlab program was developed to simulate a multilayer SPR surface, and used to assess how well antibodies can be immobilized on the sensor. The project also presents a method for making SPR surfaces from a polished glass wafer and a procedure for immobilizing antibodies on these surfaces. This procedure was used to build a biosensor for cardiac Troponin I (cTnI), and the sensor was tested. A commercially available SPR chip (Xantec SPR sensor chip CMD500l) was also tested and produced more reproducible data. However, when tested on cTnI diluted in bovine serum albumin, the sensor performed poorly. The study concludes that further experiments with different pH values and surface coatings are needed.
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