Phosphate → 2-[[2-[(2-Aminoacetyl)amino]acetyl]amino]-5-(diaminomethylideneamino)pentanoic acid

HRID 1607824Notes

Reaction equation

HRID 1607824 reaction equation
[NH2:1][CH2:2][C:3]([NH:5][CH2:6][C:7]([NH:9][C@H:10]([C:18](N[C@H](C(O)=O)CC1C=CC(O)=CC=1)=[O:19])[CH2:11][CH2:12][CH2:13][NH:14][C:15](=[NH:17])[NH2:16])=[O:8])=[O:4].P([O-])([O-])([O-])=[O:34].C([BH3-])#N.[Na+]>>[CH2:12]([CH2:13][N:14]=[C:15]([NH2:16])[NH2:17])[CH2:11][CH:10]([NH:9][C:7]([CH2:6][NH:5][C:3]([CH2:2][NH2:1])=[O:4])=[O:8])[C:18]([OH:19])=[O:34]

Participants

4

Reactant 3

HCID 1061PhosphateHCID 5003444Sodium cyanoborohydrideHCID 178512897HCID 178512897, compound2×

Product 1

HCID 3334032-[[2-[(2-Aminoacetyl)amino]acetyl]amino]-5-(diaminomethylideneamino)pentanoic acid

Procedure

Gly-Gly-Arg-Tyr*, prepared by the method of Section 9.2, was coupled to normal human IgG or LL 1151 or NS 4.1 antibody oxidized by the method of Section 12.1 infra, by incubating the antibody at a 300-fold molar excess of Gly-Gly-Arg-Tyr* in the 0.1 M phosphate buffer, pH 6.0. To reduce the imine, sodium cyanoborohydride (NaCNBH3) was added to a final concentration of 10 mM, and the reaction mixture was maintained at room temperature for 2 hours. Unreacted Gly-Gly-Arg-Tyr* was separated from theantibody conjugate by gel filtration. The sample was passed through a 5 ml Sephadex® G-50 column (Pharmacia Fine Chemicals, Piscataway, N.J.), which had been pre-coated with 1 ml of 10% BSA and run in PBS, pH 7.4. Theprotein fractions were pooled.

Workup

  1. additionwas added to a final concentration of 10 mM
  2. customUnreacted Gly-Gly-Arg-Tyr* was separated from theantibody conjugate by gel filtration

Sources & evidence

ORD record
ord-4e32bc3942c94367bd2fcaead6a28c38
Source dataset
uspto-grants-1989_09
Patent number
US04867973

Reaction notes

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