Biotin immobilization on the EGOFET surface Sample Clauses

Biotin immobilization on the EGOFET surface. In strategy #1, the commercially available poly(3-carboxypropylthiophene) (P3CPT) was activated by N-hydroxy-succinimide and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (NHS/EDC) chemistry at VTT and both biotin and maleimide groups were bound to the layers to enable a binding of avidin. A cysteine modified chimeric avidin (ChiAvd-Cys) was also bound through maleimide to the surface. Avidin with monolayer surface coverage (430 - 470 ± 30 ngcm-2) could be bound to the P3CPT- COOH activated surface. Additionally, VTT studied the possibility to immobilize ChiAvd-Cys directly by adsorption onto poly(3-hexylthiophene) (P3HT) and blocking with the polymer N-[tris(hydroxy- methyl)methyl]acrylamide - lipoic acid conjugate (pTHMMAA). Also, a new method based on photobiotin was studied by ACREO and evaluated. These two new approaches would be more compatible with the demands for printing of the avidin-antibody layer, by reducing the amount of immobilisation steps. An activation of the surface by NHS/EDC chemistry is too complicated when layers are to be printed on the semiconductor surface. In order to enable avidin coupling to the semiconductor surface, new functionalized polythiophenes that allow the grafting of biotin was synthesised at UPD (strategy #2). The biotin groups were, however, not available for a specific binding to the surface, but avidin was non-specifically adsorbed on the surface. The most promising result was obtained by a combination with strategy #1, an additional treatment of the co-polythiophene surface with a short hydrophilic blocking molecule. Biotinylated phospholipids were covalently anchored to P3HT-COOH and avidin were coupled corresponding to 150 ± 20 ng cm-2 were coupled to the layers (UNIBA - strategy #3).
Sample 1
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