title: An improved method for conjugating monoclonal antibodies with N-hydroxysulfosuccinimidyl DOTA.
abstract: A simple, water-soluble procedure for conjugation of monoclonal antibodies to 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid (DOTA) has been improved by optimizing pH, buffer, and temperature conditions for the preparation of N-hydroxysulfosuccinimidyl DOTA and its conjugation to the human/murine chimeric anti-carcinoembryonic antigen antibody cT84.66. This improved method results in a 6-fold increase in conjugation efficiency, a 3-7-fold decrease in antibody cross-linking, a more homogeneous population of conjugate species, and a 5-fold decrease in the quantities of reagents needed for conjugation. The cT84.66-DOTA conjugate was labeled to high specific activity with 111In, 90Y, 88Y, 64Cu, and 67Cu, affording near-quantitative incorporation of the majority of these radiometals. This improved conjugation procedure facilitates large-scale production and radiometal labeling of cT84.66-DOTA for clinical radioimmunotherapy trials.
authors: City of Hope Graduate Program in Biological Sciences, Department of Radioimmunotherapy, City of Hope National Medical Center, Duarte, California 91010, USA.
authors: M R
authors: MR
authors: Lewis
authors: J Y
authors: JY
authors: Kao
authors: A L
authors: AL
authors: Anderson
authors: J E
authors: JE
authors: Shively
authors: A
authors: A
authors: Raubitschek
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000912:Antibodies, Neoplasm / Q000737:chemistry*; D002021:Buffers; D002272:Carcinoembryonic Antigen / Q000276:immunology*; D020740:Chimerin Proteins / Q000737:chemistry; D006573:Heterocyclic Compounds, 1-Ring / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D018796:Immunoconjugates / Q000737:chemistry*; D007525:Isoelectric Focusing; D008670:Metals / Q000737:chemistry; D051379:Mice; D016719:Radioimmunodetection; D011868:Radioisotopes / Q000737:chemistry; D013388:Succinimides / Q000737:chemistry*; D013696:Temperature
chemical_list: D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D002021:Buffers; D002272:Carcinoembryonic Antigen; D020740:Chimerin Proteins; D006573:Heterocyclic Compounds, 1-Ring; D018796:Immunoconjugates; D008670:Metals; D011868:Radioisotopes; D013388:Succinimides; C071349:1,4,7,10-tetraazacyclododecane- 1,4,7,10-tetraacetic acid; C035761:N-hydroxysulfosuccinimide