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Contenido archivado el 2024-04-30

Multianalyte Diagnosis at the level of single Biomolecules in real time using atomic force microscopy: a new diagnostic concept

Objetivo



Summary Atomic Force Microscopy (AFM) is a new, fairly inexpensive technic, `which allows visualization of single biomolecules under physiological conditions. It also allows visualization of the interaction of biomolecules in real time. In addition it allows nanomanipulation while its theoretical limit of resolution is at the atomic scale.
Many groups in the USA and a few in Europe are presently exploring its ibiomedical capabilities. The first results are fully in accordance with the expectations based upon the above described properties of AFM, and strengthen the impression that this technic will revolutionise biomedical research in the years to come.
The target of this project is to take advantage of the resolution and 'the speed provided by AFM to a design a method to test single sera against a very large number of antigens in a very short time (minutes, probably seconds). Multiple antigens will be covalently coupled to a small area of ultra flat polyethylene or polypropylene. Antibodies can than be bound and subsequently removed without affecting the covalently bound antigens.
With the appropriate antibodies the exact position of each separate antigen will be determined using AFM. When all antigens are located on .this "antigenic surface", an antiserum to be tested is then contacted with this surface (in a flowthrough cuvette) and binding of antibodies is observed in real time at exact positions. Because the antigen at each position is known, the position at which antibodies bind will directly reveal their specificity. After the determination is done (probably a matter of seconds) antibodies are removed from the "antigenic surface" , and the next serum can be tested. At present no such multi-analyte method exist or can be envisaged otherwise. Although this project is not stretching the limits of AFM, it will nevertheless provide new diagnostic procedures for laboratories, hospitals as well as at the physicians offices.
Finally the proposed method will be the first step towards other exciting possibilities, for instance new ways to design drugs based upon diversity libraries of ultra small size or to read and write data at the molecular level. The project combines two SME's one which is a producer of AFM machines in Europe, the other is an emerging biotech company with a longstanding experience in the diagnostic field, a participant from a research institute devoted to animal disease and diagnostics in the veterinary field is included, while the last participant has pioneered the use of solid supports coupled peptides to map systematically antigenic sites of antibodies, T-cells and recently receptors, and is heavily involved in multi- analyte diagnosis using more conventional methods.

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AGRICULTURAL RESEARCH DEPARTMENT
Aportación de la UE
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15,Edelhertweg 15
8200 AB LELYSTAD
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