99mTc-labeled divalent and tetravalent CC49 single-chain Fv's

Novel imaging agents for rapid in vivo localization of human colon carcinoma

A. Goel, J. Baranowska-Kortylewicz, S. H. Hinrichs, J. Wisecarver, G. Pavlinkova, Samuel C. Augustine, D. Colcher, B. J M Booth, S. K. Batra

Research output: Contribution to journalArticle

66 Citations (Scopus)

Abstract

Radioimmunopharmaceutical agents enabling rapid high-resolution imaging, high tumor-to-background ratios, and minimal immunogenicity are being sought for cancer diagnosis and imaging. Genetic engineering techniques have allowed the design of single-chain Fv's (scFv's) of monoclonal antibodies (mAbs) recognizing tumor-associated antigens. These scFv's show good tumor targeting and biodistribution properties in vivo, indicating their potential as imaging agents when labeled with a suitable radionuclide. Methods: Divalent (sc(Fv)2) and tetravalent ([sc(Fv)2]2) scFv's of mAb CC49 were evaluated for radioimmunolocalization of LS-174T colon carcinoma xenografts in athymic mice. scFv's were radiolabeled with 99mTc by way of the bifunctional chelator succinimidyl-6-hydrazinonicotinate hydrochloride using tricine as the transchelator. The immunoreactivity and in vitro stability of the scFv's were analyzed after radiolabeling. Biodistribution and pharmacokinetic studies were performed to determine the tumor-targeting potential of the radiolabeled scFv's. Whole-mouse autoradiography illustrated the possible application of these 99mTc-labeled multivalent scFv's for imaging. Results: The radiolabeling procedure gave ≥95% radiometal incorporation, with a specific activity of >74 MBq/mg scFv. In solid-phase radioimmunoassay, both sc(Fv)2 and [sc(Fv)2]2 exhibited 75%-85% immunoreactivity, with nonspecific binding between 0.8% and 1.2%. Size-exclusion high-performance liquid chromatography showed sc(Fv)2 as a 60-kDa protein and [sc(Fv)2]2 as a 120-kDa protein. Blood clearance studies showed the elimination half-life of 99mTc-labeled sc(Fv)2 as 144 min and that of [sc(Fv)2]2 as 307 min. Whole-body clearance studies confirmed the rapid elimination of scFv's, with half-lives of 184±19 min for sc(Fv)2 and 265±39 min for [sc(Fv)2]2 (P99mTc-sc(Fv)2. 99mTc-[sc(Fv)2]2 showed a tumor uptake of 19.1±1.1%lD/g at the same time; the amount of radioactivity in the tumors was 4-fold higher than in the spleen and kidneys and 2-fold higher than in the liver. Macroautoradiography performed at 6 and 16 h after administration clearly detected the tumor with both scFv's. Conclusion: 99mTc-labeled multivalent scFv's show good tumor-targeting characteristics and high radiolocalization indices (tumor-to-back-ground ratio). These reagents, therefore, have the potential for use in clinical imaging studies of cancer in the field of nuclear medicine.

Original languageEnglish
Pages (from-to)1519-1527
Number of pages9
JournalJournal of Nuclear Medicine
Volume42
Issue number10
StatePublished - 2001
Externally publishedYes

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Single-Chain Antibodies
Colon
Carcinoma
Neoplasms
Genetic Techniques
Genetic Engineering
Nuclear Medicine
Neoplasm Antigens
Chelating Agents
Autoradiography
Heterografts
Nude Mice
Radioisotopes
Radioactivity
Radioimmunoassay
Half-Life
Proteins
Spleen
Pharmacokinetics
High Pressure Liquid Chromatography

All Science Journal Classification (ASJC) codes

  • Radiological and Ultrasound Technology

Cite this

Goel, A., Baranowska-Kortylewicz, J., Hinrichs, S. H., Wisecarver, J., Pavlinkova, G., Augustine, S. C., ... Batra, S. K. (2001). 99mTc-labeled divalent and tetravalent CC49 single-chain Fv's: Novel imaging agents for rapid in vivo localization of human colon carcinoma. Journal of Nuclear Medicine, 42(10), 1519-1527.

99mTc-labeled divalent and tetravalent CC49 single-chain Fv's : Novel imaging agents for rapid in vivo localization of human colon carcinoma. / Goel, A.; Baranowska-Kortylewicz, J.; Hinrichs, S. H.; Wisecarver, J.; Pavlinkova, G.; Augustine, Samuel C.; Colcher, D.; Booth, B. J M; Batra, S. K.

In: Journal of Nuclear Medicine, Vol. 42, No. 10, 2001, p. 1519-1527.

Research output: Contribution to journalArticle

Goel, A, Baranowska-Kortylewicz, J, Hinrichs, SH, Wisecarver, J, Pavlinkova, G, Augustine, SC, Colcher, D, Booth, BJM & Batra, SK 2001, '99mTc-labeled divalent and tetravalent CC49 single-chain Fv's: Novel imaging agents for rapid in vivo localization of human colon carcinoma', Journal of Nuclear Medicine, vol. 42, no. 10, pp. 1519-1527.
Goel, A. ; Baranowska-Kortylewicz, J. ; Hinrichs, S. H. ; Wisecarver, J. ; Pavlinkova, G. ; Augustine, Samuel C. ; Colcher, D. ; Booth, B. J M ; Batra, S. K. / 99mTc-labeled divalent and tetravalent CC49 single-chain Fv's : Novel imaging agents for rapid in vivo localization of human colon carcinoma. In: Journal of Nuclear Medicine. 2001 ; Vol. 42, No. 10. pp. 1519-1527.
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title = "99mTc-labeled divalent and tetravalent CC49 single-chain Fv's: Novel imaging agents for rapid in vivo localization of human colon carcinoma",
abstract = "Radioimmunopharmaceutical agents enabling rapid high-resolution imaging, high tumor-to-background ratios, and minimal immunogenicity are being sought for cancer diagnosis and imaging. Genetic engineering techniques have allowed the design of single-chain Fv's (scFv's) of monoclonal antibodies (mAbs) recognizing tumor-associated antigens. These scFv's show good tumor targeting and biodistribution properties in vivo, indicating their potential as imaging agents when labeled with a suitable radionuclide. Methods: Divalent (sc(Fv)2) and tetravalent ([sc(Fv)2]2) scFv's of mAb CC49 were evaluated for radioimmunolocalization of LS-174T colon carcinoma xenografts in athymic mice. scFv's were radiolabeled with 99mTc by way of the bifunctional chelator succinimidyl-6-hydrazinonicotinate hydrochloride using tricine as the transchelator. The immunoreactivity and in vitro stability of the scFv's were analyzed after radiolabeling. Biodistribution and pharmacokinetic studies were performed to determine the tumor-targeting potential of the radiolabeled scFv's. Whole-mouse autoradiography illustrated the possible application of these 99mTc-labeled multivalent scFv's for imaging. Results: The radiolabeling procedure gave ≥95{\%} radiometal incorporation, with a specific activity of >74 MBq/mg scFv. In solid-phase radioimmunoassay, both sc(Fv)2 and [sc(Fv)2]2 exhibited 75{\%}-85{\%} immunoreactivity, with nonspecific binding between 0.8{\%} and 1.2{\%}. Size-exclusion high-performance liquid chromatography showed sc(Fv)2 as a 60-kDa protein and [sc(Fv)2]2 as a 120-kDa protein. Blood clearance studies showed the elimination half-life of 99mTc-labeled sc(Fv)2 as 144 min and that of [sc(Fv)2]2 as 307 min. Whole-body clearance studies confirmed the rapid elimination of scFv's, with half-lives of 184±19 min for sc(Fv)2 and 265±39 min for [sc(Fv)2]2 (P99mTc-sc(Fv)2. 99mTc-[sc(Fv)2]2 showed a tumor uptake of 19.1±1.1{\%}lD/g at the same time; the amount of radioactivity in the tumors was 4-fold higher than in the spleen and kidneys and 2-fold higher than in the liver. Macroautoradiography performed at 6 and 16 h after administration clearly detected the tumor with both scFv's. Conclusion: 99mTc-labeled multivalent scFv's show good tumor-targeting characteristics and high radiolocalization indices (tumor-to-back-ground ratio). These reagents, therefore, have the potential for use in clinical imaging studies of cancer in the field of nuclear medicine.",
author = "A. Goel and J. Baranowska-Kortylewicz and Hinrichs, {S. H.} and J. Wisecarver and G. Pavlinkova and Augustine, {Samuel C.} and D. Colcher and Booth, {B. J M} and Batra, {S. K.}",
year = "2001",
language = "English",
volume = "42",
pages = "1519--1527",
journal = "Journal of Nuclear Medicine",
issn = "0161-5505",
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}

TY - JOUR

T1 - 99mTc-labeled divalent and tetravalent CC49 single-chain Fv's

T2 - Novel imaging agents for rapid in vivo localization of human colon carcinoma

AU - Goel, A.

AU - Baranowska-Kortylewicz, J.

AU - Hinrichs, S. H.

AU - Wisecarver, J.

AU - Pavlinkova, G.

AU - Augustine, Samuel C.

AU - Colcher, D.

AU - Booth, B. J M

AU - Batra, S. K.

PY - 2001

Y1 - 2001

N2 - Radioimmunopharmaceutical agents enabling rapid high-resolution imaging, high tumor-to-background ratios, and minimal immunogenicity are being sought for cancer diagnosis and imaging. Genetic engineering techniques have allowed the design of single-chain Fv's (scFv's) of monoclonal antibodies (mAbs) recognizing tumor-associated antigens. These scFv's show good tumor targeting and biodistribution properties in vivo, indicating their potential as imaging agents when labeled with a suitable radionuclide. Methods: Divalent (sc(Fv)2) and tetravalent ([sc(Fv)2]2) scFv's of mAb CC49 were evaluated for radioimmunolocalization of LS-174T colon carcinoma xenografts in athymic mice. scFv's were radiolabeled with 99mTc by way of the bifunctional chelator succinimidyl-6-hydrazinonicotinate hydrochloride using tricine as the transchelator. The immunoreactivity and in vitro stability of the scFv's were analyzed after radiolabeling. Biodistribution and pharmacokinetic studies were performed to determine the tumor-targeting potential of the radiolabeled scFv's. Whole-mouse autoradiography illustrated the possible application of these 99mTc-labeled multivalent scFv's for imaging. Results: The radiolabeling procedure gave ≥95% radiometal incorporation, with a specific activity of >74 MBq/mg scFv. In solid-phase radioimmunoassay, both sc(Fv)2 and [sc(Fv)2]2 exhibited 75%-85% immunoreactivity, with nonspecific binding between 0.8% and 1.2%. Size-exclusion high-performance liquid chromatography showed sc(Fv)2 as a 60-kDa protein and [sc(Fv)2]2 as a 120-kDa protein. Blood clearance studies showed the elimination half-life of 99mTc-labeled sc(Fv)2 as 144 min and that of [sc(Fv)2]2 as 307 min. Whole-body clearance studies confirmed the rapid elimination of scFv's, with half-lives of 184±19 min for sc(Fv)2 and 265±39 min for [sc(Fv)2]2 (P99mTc-sc(Fv)2. 99mTc-[sc(Fv)2]2 showed a tumor uptake of 19.1±1.1%lD/g at the same time; the amount of radioactivity in the tumors was 4-fold higher than in the spleen and kidneys and 2-fold higher than in the liver. Macroautoradiography performed at 6 and 16 h after administration clearly detected the tumor with both scFv's. Conclusion: 99mTc-labeled multivalent scFv's show good tumor-targeting characteristics and high radiolocalization indices (tumor-to-back-ground ratio). These reagents, therefore, have the potential for use in clinical imaging studies of cancer in the field of nuclear medicine.

AB - Radioimmunopharmaceutical agents enabling rapid high-resolution imaging, high tumor-to-background ratios, and minimal immunogenicity are being sought for cancer diagnosis and imaging. Genetic engineering techniques have allowed the design of single-chain Fv's (scFv's) of monoclonal antibodies (mAbs) recognizing tumor-associated antigens. These scFv's show good tumor targeting and biodistribution properties in vivo, indicating their potential as imaging agents when labeled with a suitable radionuclide. Methods: Divalent (sc(Fv)2) and tetravalent ([sc(Fv)2]2) scFv's of mAb CC49 were evaluated for radioimmunolocalization of LS-174T colon carcinoma xenografts in athymic mice. scFv's were radiolabeled with 99mTc by way of the bifunctional chelator succinimidyl-6-hydrazinonicotinate hydrochloride using tricine as the transchelator. The immunoreactivity and in vitro stability of the scFv's were analyzed after radiolabeling. Biodistribution and pharmacokinetic studies were performed to determine the tumor-targeting potential of the radiolabeled scFv's. Whole-mouse autoradiography illustrated the possible application of these 99mTc-labeled multivalent scFv's for imaging. Results: The radiolabeling procedure gave ≥95% radiometal incorporation, with a specific activity of >74 MBq/mg scFv. In solid-phase radioimmunoassay, both sc(Fv)2 and [sc(Fv)2]2 exhibited 75%-85% immunoreactivity, with nonspecific binding between 0.8% and 1.2%. Size-exclusion high-performance liquid chromatography showed sc(Fv)2 as a 60-kDa protein and [sc(Fv)2]2 as a 120-kDa protein. Blood clearance studies showed the elimination half-life of 99mTc-labeled sc(Fv)2 as 144 min and that of [sc(Fv)2]2 as 307 min. Whole-body clearance studies confirmed the rapid elimination of scFv's, with half-lives of 184±19 min for sc(Fv)2 and 265±39 min for [sc(Fv)2]2 (P99mTc-sc(Fv)2. 99mTc-[sc(Fv)2]2 showed a tumor uptake of 19.1±1.1%lD/g at the same time; the amount of radioactivity in the tumors was 4-fold higher than in the spleen and kidneys and 2-fold higher than in the liver. Macroautoradiography performed at 6 and 16 h after administration clearly detected the tumor with both scFv's. Conclusion: 99mTc-labeled multivalent scFv's show good tumor-targeting characteristics and high radiolocalization indices (tumor-to-back-ground ratio). These reagents, therefore, have the potential for use in clinical imaging studies of cancer in the field of nuclear medicine.

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