Microcontact printing of antibodies in complex with conjugated polyelectrolytes

University essay from Linköpings universitet/Institutionen för fysik, kemi och biologi

Abstract:

Microcontact printing using elastomeric stamps is a technique used in finding new and efficient ways to produce biodetection chips. Microcontact printed, with poly(dimetylslioxane) (PDMS) stamps, patterns of antibodies have been evaluated using fluorescence microscopy, imaging ellipsometry and atomic force microscopy. Fluorescent conjugated polyelectrolytes form non-covalent molecular complexes with Immunoglobulin-γ type antibodies, antigen binding to the tagged antibody result in spectroscopic shifts. Four different conjugated polyelectrolytes (POWT, POMT, PTT, PTAA) in complex with human serum albumin antibodies (aHSA) have been tested with fluorescence spectroscopy. Complexes of POWT and aHSA gave rise to thelargest wavelength shift when exposed to human serum albumin.

Several types of commercially available fluorescent antibodies and antigens were used to test the specificity of microcontact printed antibodies to different antigen solutions. Using fluorescence microscopy it could not be shown that printed antibody patterns promote specific adsorption of corresponding antigen. It is proposed however that changed surface characteristics of the substrate due to PDMS residues transferred during printing is the main driving force behind antigen adsorption.

POMT - poly (3-[(s)-5-amino-5-methoxylcarboxyl-3-oxapentyl]-2,5-thiophenylenehydrochloride)

POWT - poly (3-(s)-5-amino-5-carboxyl-3-oxapentyl]-2,5-thiophenylenehydrochloride)

PTAA - polytiophene acetic acid

PTT - poly (3-[2,5,8-trioxanonyl] thiophene)

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