A method for preparing erythropoietin (EPO) to which a compound represented by Formula 1 is coupled only to a tyrosine of EPO, comprising reacting a compound represented by Formula 1 and EPO in an aqueous solution in the presence of 1,1,3,3-tetramethylguanidine (TMG):
- wherein A is nonsubstituted or substituted C6-14 arylene or 5-20 membered nonsubstituted or substituted heteroarylene, wherein the substituted arylene and heteroarylene can be substituted with one or more selected from the group consisting of halogen, straight or branched C1-5 alkyl, straight or branched C1-5 alkoxy, ═O and —OH;
- L is any one selected from the group consisting of S, O, NR3, —NHC(═O)—, —C(═O)NH— and straight or branched C1-10 alkylene, or a combination thereof, and R3 is hydrogen or straight or branched C1-3 alkyl;
- and
- B is a biocompatible polymer, organic or inorganic fluorescent substance, or an antibody.