Abstract
We report a new generic device suitable for the investigation of biological interactions by means of Fourier Transform Infrared (FTIR) spectroscopy. The research is focused on multi reflected evanescent optical radiation through a chemically modified surface of the attenuated total reflection (ATR) element (silicon or germanium). Using a wet chemistry approach, the original method is based on grafting of a bifunctional binding molecule (N-hydroxysuccinimidyl ester forms) at the surface of the Si or the Ge crystals. The functionalized surface permits then the foundation of different types of self-assembled phospholipid membranes. The obtained sensors allow the detection, in infrared spectral domain, of any perceptible molecular interaction or structural changes. The key experimental result concerns the coagulation factor VIII(FVIII). The principle of the diagnostic is related to the ability of FFVIIImolecules to bind specifically to phosphatidylserine (PS) membrane.
Highlights
Fourier Transform Infrared (FTIR) (Fourier transform infrared) spectroscopy is one of the most powerful techniques for structural and spectral direct recognition of biological materials and their interactions [1]
Well-ordered assemblies of phospholipid films are important for the success of our binding studies of factor VIII (FVIII)
Concerning the sensors using Ge, our results show a preferential binding of the FVIII molecules on the PS membranes but the quality of the control experiment is not as good as expected on the basis of the results obtained on Si
Summary
FTIR (Fourier transform infrared) spectroscopy is one of the most powerful techniques for structural and spectral direct recognition of biological materials and their interactions [1]. The biological molecules are primarily studied in aqueous solutions. In order to analyse the FTIR spectra, the main problem one should overcome is the strong water absorption. In combination with the attenuated total reflection (ATR) method, FTIR spectroscopy eases this effect and strengthens the signal to noise ratio [2]. The Internal Reflection Element (IRE) plays a major function in the ATR system. The IRE allows the multiplication of the number of reflections, and on the other, a portion of the IR light penetrates the IRE surface and probes the response of the highly concentrated molecules lying close to the interface
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