title: Simplified method for conjugating macrocyclic bifunctional chelating agents to antibodies via 2-iminothiolane.
abstract: A one-step method for conjugating macrocyclic chelators to antibodies using the protein modification reagent 2-iminothiolane controls aggregation, maintains immunoreactivity, and produces consistent chelate/antibody ratios. Conjugation conditions have been investigated with the macrocyclic chelates 6-[p-(bromoacetamido)benzyl]-1,4,8,11-tetraazacyclotetradecane-N,N ',N",N"'-tetraacetic acid and 2-[p-(bromoacetamido)benzyl]-1,4,7,10-tetraazacyclododecane-N,N',N ",N"'-tetraacetic acid, with three different monoclonal antibodies. The bifunctional chelating agents are prepared by bromoacetylation of their amine precursors using a two-phase H2O/CHCl3 system, which improves product purity.
authors.affiliation: Chemistry Department, University of California, Davis 95616.
authors.forename: M J
authors.initials: MJ
authors.lastname: McCall
authors.forename: H
authors.initials: H
authors.lastname: Diril
authors.forename: C F
authors.initials: CF
authors.lastname: Meares
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000915:Antibody Affinity; D002614:Chelating Agents / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D003037:Cobalt Radioisotopes; D003432:Cross-Linking Reagents; D006571:Heterocyclic Compounds / Q000737:chemistry*; D007096:Imidoesters / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D002614:Chelating Agents; D003037:Cobalt Radioisotopes; D003432:Cross-Linking Reagents; D006571:Heterocyclic Compounds; D007096:Imidoesters; C046529:6-(4-bromoacetamidobenzyl)-1,4,8,11-tetraazacyclotetradecane-N,N',N'',N'''-tetraacetic acid; C063627:4-bromoacetamidobenzyl-1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid; C005905:methyl 4-mercaptobutyrimidate