HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH RSS TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Figures Only Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Cheung, N.-K. V. Articles by Larson, S. M. Search for Related Content PubMed PubMed Citation Articles by Cheung, N.-K. V. Articles by Larson, S. M.

Single-Chain Fv-Streptavidin Substantially Improved Therapeutic Index in Multistep Targeting Directed at Disialoganglioside GD2

¹ Department of Pediatrics, Memorial Sloan-Kettering Cancer Center, New York, New York
² NeoRx Corporation, Seattle, Washington
³ Department of Nuclear Medicine, Memorial Sloan-Kettering Cancer Center, New York, New York

Multistep targeting can improve the therapeutic index of antibody-based targeting, particularly relevant to pediatric tumors where acute toxicity and late effects of treatment are major concerns. Neuroblastoma is uniquely suited for such investigations because of its abundance of surface ganglioside GD2. Methods: 5F11scFv (scFv = single-chain variable fragment) was constructed from the variable regions of the heavy (V_H) and -light (V_L) chain complementary DNA (cDNA) of anti-GD2 IgM hybridoma 5F11 and ligated to full-length streptavidin cDNA for expression in Escherichia coli. Purified 5F11-scFv-streptavidin (5F11-scFv-SA) was a homotetramer and showed comparable avidity to 5F11 IgM and a 30-fold improvement over monomeric scFv. Biodistribution of 5F11-scFv-SA was studied in nude mice xenografted with neuroblastoma LAN-1. Twenty-four hours after intravenous injection of 300–900 µg 5F11-scFv-SA, 150–450 µg of a thiogalactoside-containing clearing agent, (Gal-NAc)₁₆--S-C₅H₁₀-NH-LC-N-Me-biotin (molecular weight, 8,652), were administered intravenously, followed by 2.5 µg (1.85–3.7 MBq) ¹¹¹In-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid-biotin (¹¹¹In-DOTA-biotin) intravenously 4 h later and clocked as time 0. Results: Tumor uptake (percentage of injected dose per gram [%ID/g]) at 2 h was 7 %ID/g and decayed with a half-life of 72 h, whereas blood %ID/g rapidly decreased to <1/500 of that of tumor after the first 24 h. The tumor-to-nontumor (T/NT) ratio at 72 h was high (median, 106; range, 3.4 [kidney] to 1,660 [blood]). When the area under the radioactivity curve was computed, the T/NT organ ratio was favorable (4.8 for kidney and 162 for blood). When human and murine tumors were surveyed, the T/NT ratio of ¹¹¹In-DOTA-biotin uptake correlated with their levels of GD2 expression as assayed by flow cytometry. Biotinylated polypeptides (bovine serum albumin and vasointestinal peptides) achieved selective tumor targeting when the multistep strategy was applied. Conclusion: Improvement in the T/NT ratio using pretargeting strategy may increase the efficacy and safety of scFv-based approaches in cancer therapy. Additionally, since biotinylated polypeptides can be rendered tumor selective, a large repertoire of agents can potentially be explored.

Key Words: single-chain variable fragment • scFv-streptavidin • multistep targeting • dosimetry • biodistribution

This article has been cited by other articles:

				J. Hu, X. Huang, C.-C. Ling, D. R. Bundle, and N.-K. V. Cheung Reducing Epitope Spread during Affinity Maturation of an Anti-Ganglioside GD2 Antibody J. Immunol., November 1, 2009; 183(9): 5748 - 5755. [Abstract] [Full Text] [PDF]

				Y. Lv, N.-K. V. Cheung, and B. M. Fu A Pharmacokinetic Model for Radioimmunotherapy Delivered Through Cerebrospinal Fluid for the Treatment of Leptomeningeal Metastases J. Nucl. Med., August 1, 2009; 50(8): 1324 - 1331. [Abstract] [Full Text] [PDF]

				R. M. Sharkey, H. Karacay, S. Litwin, E. A. Rossi, W. J. McBride, C.-H. Chang, and D. M. Goldenberg Improved Therapeutic Results by Pretargeted Radioimmunotherapy of Non-Hodgkin's Lymphoma with a New Recombinant, Trivalent, Anti-CD20, Bispecific Antibody Cancer Res., July 1, 2008; 68(13): 5282 - 5290. [Abstract] [Full Text] [PDF]

				D. M. Goldenberg, R. M. Sharkey, G. Paganelli, J. Barbet, and J.-F. Chatal Antibody Pretargeting Advances Cancer Radioimmunodetection and Radioimmunotherapy J. Clin. Oncol., February 10, 2006; 24(5): 823 - 834. [Abstract] [Full Text] [PDF]

				G. J. Forster, E. B. Santos, P. M. Smith-Jones, P. Zanzonico, and S. M. Larson Pretargeted Radioimmunotherapy with a Single-Chain Antibody/Streptavidin Construct and Radiolabeled DOTA-Biotin: Strategies for Reduction of the Renal Dose J. Nucl. Med., January 1, 2006; 47(1): 140 - 149. [Abstract] [Full Text] [PDF]

				D. J. Buchsbaum, M.B. Khazaeli, D. B. Axworthy, J. Schultz, T. R. Chaudhuri, K. R. Zinn, M. Carpenter, and A. F. LoBuglio Intraperitoneal Pretarget Radioimmunotherapy with CC49 Fusion Protein Clin. Cancer Res., November 15, 2005; 11(22): 8180 - 8185. [Abstract] [Full Text] [PDF]

				S. Modak and N.-K. V. Cheung Antibody-Based Targeted Radiation to Pediatric Tumors J. Nucl. Med., January 1, 2005; 46(1_suppl): 157S - 163S. [Abstract] [Full Text] [PDF]