| 摘要 |
A method of producing a reagent for use in a conjugation reaction between a first reactant containing a carboxyl group or a phosphate group and a second reactant containing an amine group, comprises: a) forming a solution or suspension of:i) a carboxyl-activating or phosphate-activating non-enzymatic substance capable of acting on a carboxyl group- or phosphate group-containing moiety to facilitate the formation of an amide bond between a carboxyl group and an amine group or a phosphoramidate bond between a phosphate group and an amine group, andii) a first reactant or a second reactant; andb) drying the solution or suspension. The resulting reagent may be useful in conjugation reactions and preferably includes no NHS (N-Hydroxysuccinimide). The activating substance may be a carbodiimide (e.g. 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide (EDC, EDAC) or N-cyclohexyl-N'-(P-[N-methylmorpholino]ethyl)carbodiimide (CMC)) or Woodward's reagent K (WRK). Step (b) may comprise freeze drying. For example: dextran-coated latex microparticles are mixed with EDC, buffer and trehalose and lyophilized; upon reconstitution, conjugation to goat anti-rabbit IgG is performed. Conjugations involving Q dots, fluorescent dyes, proteins and gold particles are exemplified. |
