Disulfide bond-targeted radiolabeling

Tumor specificity of a streptavidin-biotinylated monoclonal antibody complex

Renato B. Del Rosario, Richard L. Wahl

Research output: Contribution to journalArticle

Abstract

A site-specific labeling method was developed in which sulfhydryl groups of a murine IgG2a anti-ovarian monoclonal antibody, 5G6.4, were biotinylated with N-iodoacetyl-N′-biotinylhexylenediamine (Compound 1) following partial reduction of disulfide bonds with dithiothreitol. Reaction of 1-alkylated 5G6.4 with 125I-streptavidin gave immunoreactive streptavidin-1-biotinylated complexes. Radio-fast protein liquid chromatography data were consistent with the formation of a stable monovalent streptavidin-half-antibody complex as the major species. In vivo specific localization of these radioantibody conjugates to human tumor xenografts of ovarian carcinoma was confirmed by a comparative biodistribution study in nude mice using as a control the nonspecific 125I-streptavidin-1-alkylated UPC-10 (an irrelevant IgG2a monoclonal antibody) complex prepared analogously as described above. Tumor uptake for radiolabeled 5G6.4 [0.279±0.041 % (SE) kg injection dose/g) was significantly greater [P <0.025] than for UPC-10 [0.165±0.027% kg injection dose/g]. The tumor:blood ratio (738±1.285) for 5G6.4 was ≈3 times that for UPC-10 (2.48 ± 0.708, P <0.01). This sulfhydryl site-directed approach demonstrated that reduced disulfides of monoclonal antibodies are viable sites for attaching labels without significant loss of in vitro and in vivo immunoreactivity.

Original languageEnglish (US)
JournalCancer Research
Volume50
Issue number3 SUPPL.
StatePublished - 1990
Externally publishedYes

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Streptavidin
Disulfides
Monoclonal Antibodies
Neoplasms
Injections
Dithiothreitol
Radio
Heterografts
Nude Mice
Liquid Chromatography
Carcinoma
Antibodies
Proteins

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Disulfide bond-targeted radiolabeling : Tumor specificity of a streptavidin-biotinylated monoclonal antibody complex. / Del Rosario, Renato B.; Wahl, Richard L.

In: Cancer Research, Vol. 50, No. 3 SUPPL., 1990.

Research output: Contribution to journalArticle

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abstract = "A site-specific labeling method was developed in which sulfhydryl groups of a murine IgG2a anti-ovarian monoclonal antibody, 5G6.4, were biotinylated with N-iodoacetyl-N′-biotinylhexylenediamine (Compound 1) following partial reduction of disulfide bonds with dithiothreitol. Reaction of 1-alkylated 5G6.4 with 125I-streptavidin gave immunoreactive streptavidin-1-biotinylated complexes. Radio-fast protein liquid chromatography data were consistent with the formation of a stable monovalent streptavidin-half-antibody complex as the major species. In vivo specific localization of these radioantibody conjugates to human tumor xenografts of ovarian carcinoma was confirmed by a comparative biodistribution study in nude mice using as a control the nonspecific 125I-streptavidin-1-alkylated UPC-10 (an irrelevant IgG2a monoclonal antibody) complex prepared analogously as described above. Tumor uptake for radiolabeled 5G6.4 [0.279±0.041 {\%} (SE) kg injection dose/g) was significantly greater [P <0.025] than for UPC-10 [0.165±0.027{\%} kg injection dose/g]. The tumor:blood ratio (738±1.285) for 5G6.4 was ≈3 times that for UPC-10 (2.48 ± 0.708, P <0.01). This sulfhydryl site-directed approach demonstrated that reduced disulfides of monoclonal antibodies are viable sites for attaching labels without significant loss of in vitro and in vivo immunoreactivity.",
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