Abstract: Methods and kits for structurally analyzing carbohydrate molecules are taught. Carbohydrates for analysis are derivatized (preferably methylated) and then hydrolyzed into constituent monosaccharides. The derivatized monosaccharides are then labeled by a fluorophore and separated from one another by electrophoresis. The identity of derivatized monosaccharides is established by comparison with identification standards. The electrophoresis separation patterns may be visualized by a charged coupled device camera or photographically.

Abstract: The subject invention provides methods and kits for conveniently diagnosing various physiological conditions that produce altered levels of specific carbohydrates, known as diagnostic carbohydrates. Measurement of the levels of the diagnostic carbohydrates is performed by fluorophore assisted carbohydrate electrophoresis. Physiological conditions of particular interest that may be ascertained by the subject invention include carbohydrate metabolism diseases, autoimmune diseases, neoplasia, toxic chemical exposure and microbial infections. Fluorophore assisted carbohydrate electrophoresis diagnosis may be applied to various patient specimens, including blood, urine and skin.

Abstract: The subject invention provides methods and kits for detecting increased bone resorption in patients. These methods and kits use chondroitin sulfate, a glycosaminoglycan, as a marker for bone resorption. Levels of chondroitin sulfate are detected in samples of urine or serum taken from patients. Measurements of chondroitin sulfate levels are made by methods such as fluorophore-assisted carbohydrate electrophoresis (FACE). Knowledge of increased bone resorption rates is useful in diagnosis of bone disorders such as osteoporosis.

Abstract: An apparatus for fluorescence of a biological sample includes a light illumination source, an ambient light-excluding optical chamber, a sample holder, a two-dimensional array charge coupled device ("CCD") sensor providing for on-chip signal integration of each pixel of the array, and an analog/digital converter for digitizing the CCD signal. Signal acquisition time from the array is varied so that the integrated signal from each pixel is below its saturation level. A darkfield value for each pixel is acquired and subtracted from the illuminated signal from that pixel so as to yield a darkfield corrected signal. The fluorescence value for each pixel can be compensated for variations in excitation energy reaching different sample areas. Image readout is through a single channel output.

Abstract: The present invention relates to an electrophoresis media comprising a gel and a compound capable of restricting the diffusion of water within the gel. Diffusion restricting compounds useful in the present invention include but are not limited to polyols, polymeric alcohols, polysaccharides, polyoxyethylene ethers, polyamines, polypeptides, gums, zwiterionic detergents and mixtures thereof. The present method relates to any electrophoresis media and includes but is not limited to polyacrylamide, agarose, starch, cellulose acetate and sepharose. A further embodiment of the present invention relates to a stacking gel wherein a diffusion restricting compound is used to prevent diffusion of water between the stack gel and resolving gel. The addition of a diffusion restricting compound to the stacking gel has the particular advantage of causing the stacking gel to become opaque, thereby facilitating the addition of samples to the gel.

Abstract: Polynucleotide sequences encoding enzymes that possess .alpha.2-3 neuraminidase activity are provided. Of particular interest are polynucleotide sequences encoding an enzyme having .alpha.2-3 neuraminidase activity and naturally produced by the bacteria Streptococcus pneumoniae. Recombinant DNA expression of enzymes possessing .alpha.2-3 specific neuraminidase activity is also described, including methods, recombinant host cells and a genetic construction. The invention also provides a purified enzyme having .alpha.2-3 neuraminidase activity from Streptococcus pneumoniae, wherein the enzyme is isolated from S. pneumoniea cultures. Another aspect of this invention is to provide methods of isolating genes encoding enzymes having neuraminidase activity, particularly .alpha.2-3 neuraminidase activity. The gene isolation methods of the invention comprise the step of labeling a hybridization probe derived from the neuraminidase coding portion of plasmid pND-1.

Abstract: Various assays for the detection and quantitation of carbohydrate-interacting proteins are disclosed. Carbohydrate-interacting proteins include carbohydrate modifying enzymes and carbohydrate binding proteins. The assays employ a fluorophore-labelled carbohydrate substrate that either is capable of being structurally modified by the carbohydrate-modifying enzyme being assayed or changes its electrophoretic gel migration tube upon interaction with a carbohydrate-binding protein. Methods for cloning genes encoding carbohydrate-modifying enzymes are also disclosed.

Abstract: The subject invention provides methods and kits for conveniently diagnosing various physiological conditions that produce altered levels of specific carbohydrates, known as diagnostic carbohydrates. Measurement of the levels of the diagnostic carbohydrates is performed by fluorophore assisted carbohydrate electrophoresis. Physiological conditions of particular interest that may be ascertained by the subject invention include carbohydrate metabolism diseases, autoimmune diseases, neoplasia, toxic chemical exposure and microbial infections. Fluorophore assisted carbohydrate electrophoresis diagnosis may be applied to various patient specimens, including blood, urine and skin.