Fabrication and characterisation of spin coated oxidised PMMA to provide a robust surface for on-chip assays.

A method to prepare a highly stable carboxylic acid functional surface on various substrates for use in bioassays is reported. A thin layer of a poly(methylmethacrylate) (PMMA) is achieved by spin coating dissolved PMMA onto a variety of underlying substrates at various thicknesses in a range of c. 5-27 nm. Varying the PMMA concentration, time and spinning speed controls the thickness of the spin coated layer. The root-mean-squared roughness values of the spin coated PMMA are less than 1.5 nm, resulting in smooth and uniform layer. Substrate functionalisation is carried out by either ultraviolet/ozone (UV/O3) or oxygen plasma oxidation. Both techniques result in initially stable, highly functional films as demonstrated by the covalent attachment of amino-modified oligonucleotides, however longevity studies comparing the stability of films attached following oxidative activation show better stability for UV/O3 activated substrates when compared to oxygen plasma activated substrates. PMMA films activated by UV/O3 yield highly stable (for up to 24 days) functional surfaces that retain immobilised biomolecules after several extended wash steps. In contrast, films attached to surfaces pre-treated with oxygen plasma discharge lose their functionality within 5 days of oxidation. Direct DNA and sandwich antibody assays were successfully demonstrated on the UV/O3 functionalised surfaces, showing a low level of non-specific binding. Furthermore, the quenching of fluorescently labelled biomolecules bound to PMMA-coated gold-coated slides is shown to be dependent on the PMMA thickness, indicating potential usage in surface-plasmon resonance-based assays.

Crystal structure and solid state 13C NMR analysis of N-(methyl 3,4,6-tri-O-acetyl-alpha, and beta-D-glucopyranosid-2-yl)-oxamide derivative of p-chloroaniline, N,N-diethylamine, N-methylaniline and N-ethylaniline.

The X-ray diffraction analysis of N-(methyl 3,4,6-tri-O-acetyl-alpha-D-glucopyranosid-2-yl)-N'-p-chlorophenyloxamide (1), N-(methyl 3,4,6-tri-O-acetyl-alpha-D-glucopyranosid-2-yl)-N',N'-diethyloxamide (2), N-acetyl, N-(methyl 3,4,6-tri-O-acetyl-beta-D-glucopyranosid-2-yl), N'-methyl, N'-phenyloxamide (3), N-acetyl, N-(methyl 3,4,6-tri-O-acetyl-beta-D-glucopyranosid-2-yl), N'-ethyl, N'-phenyloxamide (4) was performed. It was found that the oxamide group in compounds 1-4 can be characterized as two structurally independent amides because there is no pi conjugation across the oxalyl OC-CO bond. Only the oxamide group of 1 is planar and adopts trans conformation stabilized as two intramolecular N-H...O hydrogen bonds.