Government of the United States of America, As Represented by the Secretary of Commerce

Other

National Institute of Standards and Technology

Michael J Tarlov

David Judson Ross

Laurie E Locascio

Karin M Balss

Tonya M Herne

Todd A Morris

Keith H McKenney

Germarie Sanchez-Pomales

Rebecca A Zangmeister

Susan L R Barker

A method of characterizing glycans attached to glycoproteins is disclosed herein. The method comprises a first step of immobilizing the glycoproteins on colloidal particles forming glycoprotein/colloidal particles. The glycans on the glycoproteins may then be characterized, for example the composition and/or structure of glycans may be characterized or the glycans attached to proteins may be identified. Characterization may be accomplished by either binding the glycoprotein/colloidal particles with one or more binding agents and assessing the aggregation of the glycoprotein/colloidal particles or by cleaving glycans from the glycoprotein/colloidal particles with a cleaving agent and analyzing the glycans.

Method of characterizing glycans attached to glycoproteins

A method is provided for observing mixing interactions and reactions of two materials in a fluid. The method in one form provides for concentrating by balancing electrophoretic velocities of a material against the bulk flow of fluid in the presence of a temperature gradient. Using an appropriate fluid, the temperature gradient can generate a corresponding gradient in the electrophoretic velocity of the material so that the electrophoretic and bulk velocities sum to zero at a unique position and the material will be focused at that position. A second material can then be introduced into the fluid and allowed to move through and interact with the focused band of the first material. Products of the interaction can then be detected as they are focused at a different position along the gradient. The method can be adapted to study the temperature dependence of the molecular interaction.

Mixing reactions by temperature gradient focusing

A microchannel device is provided with a plastic substrate having a microchannel formed therein. Polyelectrolyte multilayers are disposed along selected surfaces of the microchannel. The polyelectrolyte layers comprise alternating net positively charged layers and net negatively charged layers. A microchannel lid has a surface facing the microchannel. In making the microchannel device, selected surfaces of the microchannel are alternatively exposed to solutions comprising positively charged polyelectrolytes and negatively charged polyelectrolytes to form the desired number of polyelectrolyte layers.

Polyelectrolyte derivatization of microfluidic devices

In one embodiment, the present invention provides a biopolymer-containing monolayer comprising: thiol-derivatized biopolymers and organic thiols bound to a metal substrate. In another embodiment, the present invention provides a method for forming this biopolymer-containing monolayer. Preferably, the biopolymers are single-stranded DNA probes.

Surface immobilization of biopolymers

A process for creating a two dimensional spacial distribution pattern of ferent thiolate molecules on a substrate by illuminating a surface of a self-assembled monolayer of a first thiolate compound in the presence of oxygen with high frequency electromagnetic radiation distributed according to a desired pattern, and subsequently immersing the illuminated substrate in a solution of a second thiolate compound so that molecules of the first thiolate compound in illuminated areas of the monolayer are exchanged for molecules of said second thiolate compound; and a patterned biomolecular composite formed of a substrate which forms a self-assembled thiolate monolayer when immersed in a solution of a thiolate forming compound, a thiolate monolayer deposited on the substrate and composed of patterned areas of first and second thiolate compounds, respectively, the first thiolate compound having an affinity for specifically or nonspecifically adsorbing a biological molecule, and the second thiolate compound having essentially no affinity for the biological molecule, and at least one biological material adsorbed in a corresponding pattern on the patterned areas of the first thiolate compound in the thiolate monolayer.

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Bethesda, MD, United States of America

Michael J Tarlov