Efficient conjugation of DTPA to an IgM monoclonal antibody in ascites fluid

Berma M. Kinsey, Roger M. Macklis, James M. Ferrara, Warren W. Layne, Steven J. Burakoff, Amin I. Kassis

Research output: Contribution to journalArticlepeer-review

8 Scopus citations


We have developed a simple, rapid, and reproducible method for conjugating the bifunctional metal chelator diethylenetriaminepentaacetic acid (DTPA) to an IgM monoclonal antibody (MoAb) without first isolating the MoAb from the ascites fluid. Treatment of the protein mixture in the ascites fluid with cyclic DTPA anhydride (cDTPAA) followed by HPLC purification on a size exclusion column allowed isolation of the DTPA-IgM conjugate which could then be labeled with 111 In ≥80% yield. Over the range of total protein concentrations used (11-44 mg/mL), the number of DTPA molecules per molecule of IgM was approximately one-half the molar ratio of cDTPAA to total protein. We have used this method to prepare an 111In labeled anti-Thy 1.2 IgM, a MoAb with specificity for a murine cell-surface antigen found on normal and malignant T cells and neuroectodertmal tissues. Analysis of the DTPA-IgM conjugate prior to and after 111In labeling using indirect immunofluorescence flow cytometry and a target-cell binding assay showed that the antigen specificity of this anti-Thy 1.2 MoAb is not substantially altered by the presence of up to 8 DTPA molecules per IgM molecule.

Original languageEnglish
Pages (from-to)285-292
Number of pages8
JournalNuclear Medicine and Biology
Issue number3
StatePublished - 1988
Externally publishedYes


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