The preparation of DTPA-coupled antibodies radiolabeled with metallic radionuclides: an improved method.
Isotopes of iodine are often employed as radiolabels for antibodies used in radioimmunodetection studies in which tumor localization is determined by external imaging. Because of drawbacks associated with the use of these isotopes, alternative labeling methods have been considered; such as covalently attaching strong chelators so that the coupled protein may be radiolabeled with metallic radionuclides by chelation. We have developed a method of coupling the strong chelator diethylenetriaminepentaacetic acid (DTPA) which is simple, efficient, and superior to reported methods. Using the cyclic anhydride, coupling to IgG antibody is about 75% efficient and is completed in less than 1 min at neutral pH. Because the concentration of hydrolytic products is small, the coupled protein is rapidly purified for use or storage. Labeling of the protein is also accomplished rapidly and the labeled product has been shown to be stable both in vitro and in vivo.
D000812:Anhydrides; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002272:Carcinoembryonic Antigen / Q000276:immunology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000032:analysis / Q000378:metabolism*; D007204:Indium; D007553:Isotope Labeling / Q000379:methods* / Q000592:standards; D046911:Macromolecular Substances; D051379:Mice; D008815:Mice, Inbred Strains; D004369:Pentetic Acid; D011868:Radioisotopes; D012709:Serum Albumin / Q000276:immunology; D013997:Time Factors
D000812:Anhydrides; D000911:Antibodies, Monoclonal; D002272:Carcinoembryonic Antigen; D007074:Immunoglobulin G; D046911:Macromolecular Substances; D011868:Radioisotopes; D012709:Serum Albumin; D007204:Indium; D004369:Pentetic Acid