Chembiomed, Ltd.

Alberta Research Council Incorporated

Glyko, Inc.

M Abdul Mazid

Mohammed A Kashem

Andre P Venot

Nai-Hong Li

John C Klock

Frank M Unger

Paul Bird

Disclosed are methods for the enzymatic synthesis of alpha-sialylated oligosaccharide glycosides. Specifically, in the disclosed methods, sialyltransferase is activated to transfer an analogue of sialic acid, employed as its CMP-nucleotide derivative, to an oligosaccharide glycoside. The analogue of sialic acid and the oligosaccharide employed in this method are selected to be compatible with the sialyltransferase employed.

Methods for the enzymatic synthesis of alpha-sialylated oligosaccharide

The subject invention provides novel methods and apparatus for synthesizing carbohydrates of a desired structure. The subject synthesis methods include using carbohydrage-modifying enzymes to perform individual synthesis reaction steps and electrophoresis to purify the reaction products for possible further use as substrates in additional synthesis reactions. The subject invention also provides novel methods and apparatus for detecting carbohydrates of interest. The methods and apparatus for synthesizing or detecting carbohydrates involve the use of carbohydrate-modifying enzymes immobilized in a reaction matrix gel and fluorophore labelled substrates that may be removed from the reaction matrix gel by electrophoresis and subsequently analyzed or transferred to another reaction gel matrix. The subject invention also provides methods for detecting and quantitating specific carbohydrate-modifying enzymes that might be present in a sample for analysis.

Rapid synthesis and analysis of carbohydrates

A method of coating chromatographic particulate supports to provide a biocompatible outer layer of synthetic membrane-type film which prevents the release of fines but permits the adsorption of components to an affinity ligand is described. The matrix is provided with a membrane-type coating, which prevents leaching of fines, with a pore size of at least 20 angstroms. The coating is applied to a solid particulate under conditions where an integral membrane coat will be formed. It may also be necessary to control the size and number of pores of the membranes by treating a suspension of the solid support in a solvent which contains 0.1-1% of the support weight of a biocompatible polymer along with 0.5-5% of the weight of said polymer of a dissolved compatible pore-controlling component. The solvent is then removed from the suspension and the membrane-coated material used in extracorporeal treatment of body fluids or in other chromatographic techniques. The coating process can be conducted before or after the particulate support is functionalized and/or derivatized.

Affinity supports for hemoperfusion

A method to prepare uniform populations of macroscopic beads containing pores of at least 0.1 microns diameter is described. The beads consist essentially of a copolymer of a monoethylenically unsaturated monomer and a crosslinking polyethylenically unsaturated monomer, said resulting copolymer optionally being derivatized with functional groups, linking moieties, and/or affinity ligands. The macroporous beads are obtained by utilizing, as porogen in the polymerization reaction, porous inorganic particles which have been preabsorbed with a blowing agent. The blowing agent is not activated until after the polymerization takes place; when the blowing agent is activated, the organic polymer which isolates the inorganic porogen particles from each other in the bead is destroyed. Upon subsequent dissolution of the inorganic porogen, a network of pores throughout the organic copolymer-formed particle is formed. The resulting beads are useful in a variety of chromatographic, analytical and biomedical techniques.

Macroporous particles as biocompatible chromatographic supports

Specific sialyl transferases (ST) are important enzymes in the manufacture of carbohydrate moieties useful for pharmacological purposes and for diagnostic and clinical procedures. The invention offers simplified methods to isolate highly specific forms of these ST enzymes by taking advantage of the affinity of the ST for its acceptor substrate in the presence of the sialyl-CMP analogs, such as CDP. Specifically exemplified are isolation of the alpha 2,3-ST, specific for Le.sup.c or LacNAc, and alpha 2,6-ST specific for LacNAc.

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Edmonton, Canada

M Abdul Mazid