Conjugation reactions

摘要

We describe methods that allow either carbodiimides or other carboxyl-reactive substances to be mixed with solutions of carboxylic acids or phosphates or amines or combinations thereof, so as to form a homogeneous mixture which is then dried, preferably in a freeze drying process. The mixture is then contacted with an entity, which preferably involves the dissolution of the mixture with a buffered solution of the entity, so as to initiate a conjugation reaction between the entity and a component in the mixture.

主权项

1. A method of producing a reagent for use in a conjugation reaction comprising
a) forming a homogeneous solution or suspension of
i) a carboxyl-activating or phosphate-activating non-enzymatic substance comprising 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), N-Cyclohexyl-N′-(β-[N-methylmorpholino]ethyl)carbodiimide (CMC), 1-ethyl-3-(4-azonia-4,4-dimethylpentyl)carbodiimide (EAC), 2,2-dichloro-5-(2-phenylethyl)-4-(trimethylsilyl)-3-furanone (DPTF), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMT-MM), or N-ethyl-5-phenylisoxazolium-3′-sulphonate (Woodwards reagent K (WRK)), wherein the substance is capable of acting on a carboxyl group- or phosphate group-containing moiety to facilitate formation of (1) an amide bond between a carboxyl group and an amine group or (2) a phosphoramidate bond between a phosphate group and an amine group, andii) a first reactant containing a carboxyl group or a phosphate group, or a second reactant containing an amine group; and b) drying the solution or suspension to produce the reagent comprising a homogeneous mixture of i) and ii).