ACS Publications. Most Trusted. Most Cited. Most Read
My Activity

Figure 1Loading Img

Chemical Modification to Reduce Renal Uptake of Disulfide-Bonded Variable Region Fragment of Anti-Tac Monoclonal Antibody Labeled with 99mTc

View Author Information
Nuclear Medicine Department, Warren G. Magnuson Clinical Center, Building 21, Room 136, and Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892
Cite this: Bioconjugate Chem. 1999, 10, 3, 447–453
Publication Date (Web):March 20, 1999
Copyright © Not subject to U.S. Copyright. Published 1999 American Chemical Society

    Article Views





    Other access options


    The anti-Tac disulfide-bonded variable region fragment (dsFv) is a genetically engineered, 25 kDa, murine monoclonal antibody fragment that recognizes the α subunit of the interleukin-2 receptor (IL-2Rα). The dsFv radiolabeled with the tetrafluorophenyl ester (TFP) of [99mTc]mercaptoacetyltriglycine ([99mTc]MAG3-TFP) showed rapid tumor uptake and fast blood clearance in mice, resulting in high tumor-to-nontumor background ratios. However, its high renal uptake was a problem. In this study, we tested the effect of lowering the isoelectric point (pI) of dsFv to <9.3 on renal and tumor uptake. To lower the pI, dsFv was acylated simultaneously with both [99mTc]MAG3-TFP and TFP-glycolate. The acylation of dsFv decreased its pI and its immunoreactivity inversely proportional to the molar ratio of TFP-glycolate to dsFv, whereas the conjugation of [99mTc]MAG3-TFP alone did not. When biodistribution studies were performed in nude mice, the effect of the lowered pI was reflected primarily in decreased kidney uptake and whole-body retention, with its highest effect seen at the earliest time point (15 min) after injection. In tumor-bearing nude mice, glycolated [99mTc]MAG3-dsFv with a pI range of 4.9 to 6.5 accumulated selectively into IL-2 receptor-positive SP2/Tac tumor similar to that of the control [125I]dsFv labeled by the Iodo-Gen method, whereas its renal uptake was 25% of [125I]dsFv at 15 min. At 90 min, the ratios of tumor to receptor-negative SP2/0 tumor, liver, kidney, stomach, and blood had peaked at 10.9, 8.5, 0.3, 5.0, and 6.2, respectively, for the glycolated [99mTc]MAG3-dsFv. The corresponding ratios for [125I]dsFv were 3.7, 5.0, 0.1, 1.5, and 2.1, respectively.

    Read this article

    To access this article, please review the available access options below.

    Get instant access

    Purchase Access

    Read this article for 48 hours. Check out below using your ACS ID or as a guest.


    Access through Your Institution

    You may have access to this article through your institution.

    Your institution does not have access to this content. You can change your affiliated institution below.

     Nuclear Medicine Department.

     Laboratory of Molecular Biology.


     To whom correspondence should be addressed. Phone:  (301) 496-1426. Fax:  (301) 402-4548. E-mail:  [email protected].

    Cited By

    This article is cited by 17 publications.

    1. C. Andrew Boswell, Devin B. Tesar, Kiran Mukhyala, Frank-Peter Theil, Paul J. Fielder, and Leslie A. Khawli . Effects of Charge on Antibody Tissue Distribution and Pharmacokinetics. Bioconjugate Chemistry 2010, 21 (12) , 2153-2163.
    2. Tomoya Uehara, Daisuke Ishii, Tomoe Uemura, Hiroyuki Suzuki, Tomoko Kanei, Kyoko Takagi, Masashi Takama, Masahiro Murakami, Hiromichi Akizawa and Yasushi Arano . γ-Glutamyl PAMAM Dendrimer as Versatile Precursor for Dendrimer-Based Targeting Devices. Bioconjugate Chemistry 2010, 21 (1) , 175-181.
    3. Frank‐Jan Warnders, Marjolijn N. Lub‐de Hooge, Elisabeth G. E. de Vries, Jos G. W. Kosterink. Influence of protein properties and protein modification on biodistribution and tumor uptake of anticancer antibodies, antibody derivatives, and non‐Ig scaffolds. Medicinal Research Reviews 2018, 38 (6) , 1837-1873.
    4. Babak Behnam Azad, Vanessa Rota, Lihai Yu, Rebecca Mcgirr, André H. St. Amant, Ting-Yim Lee, Savita Dhanvantari, Leonard G. Luyt. Synthesis and Evaluation of Optical and PET GLP-1 Peptide Analogues for GLP-1R Imaging. Molecular Imaging 2015, 14 (1) , 1-16.
    5. Antony Constantinou, Chen Chen, Mahendra P. Deonarain. Polysialic Acid and Polysialylation to Modulate Antibody Pharmacokinetics. 2012, 95-115.
    6. Gaurav Malviya, Calogero D'Alessandria, Elena Bonanno, Vladimir Vexler, Roberto Massari, Carlo Trotta, Francesco Scopinaro, Rudi Dierckx, Alberto Signore. Radiolabeled Humanized Anti-CD3 Monoclonal Antibody Visilizumab for Imaging Human T-Lymphocytes. Journal of Nuclear Medicine 2009, 50 (10) , 1683-1691.
    7. Hiromichi Akizawa, Tomoya Uehara, Yasushi Arano. Renal uptake and metabolism of radiopharmaceuticals derived from peptides and proteins. Advanced Drug Delivery Reviews 2008, 60 (12) , 1319-1328.
    9. Surinder K Batra, Maneesh Jain, Uwe A Wittel, Subhash C Chauhan, David Colcher. Pharmacokinetics and biodistribution of genetically engineered antibodies. Current Opinion in Biotechnology 2002, 13 (6) , 603-608.
    10. Insook Kim, Hisataka Kobayashi, Tae M Yoo, Meyong-kon Kim, Nhat Le, Eui-sik Han, Q-C Wang, Ira Pastan, Jorge A Carrasquillo, Chang H Paik. Lowering of pI by acylation improves the renal uptake of 99mTc-labeled anti-Tac dsFv: effect of different acylating reagents. Nuclear Medicine and Biology 2002, 29 (8) , 795-801.
    11. Yubin Miao, Nellie K. Owen, Donna Whitener, Fabio Gallazzi, Timothy J. Hoffman, Thomas P. Quinn. In vivo evaluation of 188 Re‐labeled alpha‐melanocyte stimulating hormone peptide analogs for melanoma therapy. International Journal of Cancer 2002, 101 (5) , 480-487.
    12. Lin Li, Tove Olafsen, Anne-Line Anderson, Anna Wu, Andrew A. Raubitschek, John E. Shively. Reduction of Kidney Uptake in Radiometal Labeled Peptide Linkers Conjugated to Recombinant Antibody Fragments. Site-Specific Conjugation of DOTA-Peptides to a Cys-Diabody. Bioconjugate Chemistry 2002, 13 (5) , 985-995.
    13. Meyoung-kon Kim, Hyeh-Jean Jeong, Chih-Hao K Kao, Zhengsheng Yao, David S Paik, Jae Eun Pie, Hisataka Kobayashi, Thomas A Waldmann, Jorge A Carrasquillo, Chang H Paik. Improved renal clearance and tumor targeting of 99m Tc-labeled anti-Tac monoclonal antibody Fab by chemical modifications. Nuclear Medicine and Biology 2002, 29 (2) , 139-146.
    14. Hiromichi Akizawa, Yasushi Arano, Masaki Mifune, Akimasa Iwado, Yutaka Saito, Takahiro Mukai, Tomoya Uehara, Masahiro Ono, Yasushi Fujioka, Kazuma Ogawa, Yoshiaki Kiso, Hideo Saji. Effect of molecular charges on renal uptake of 111 In-DTPA-conjugated peptides. Nuclear Medicine and Biology 2001, 28 (7) , 761-768.
    15. C. W. Park, N. Sato, B. C. Shin, K. J. Wong, L. S. Park, Z. Yao, J. A. Carrasquillo, C. H. Paik. Dendrimer‐based biotin radiopharmaceutical: Chemical modifications improved in vivo clearance property. Journal of Labelled Compounds and Radiopharmaceuticals 2001, 44 (S1)
    16. Chien-Tsun Kuan, Carol J. Wikstrand, Gary Archer, Richard Beers, Ira Pastan, Michael R. Zalutsky, Darell D. Bigner. Increased binding affinity enhances targeting of glioma xenografts by EGFRvIII-specific scFv. International Journal of Cancer 2000, 88 (6) , 962-969.;962::AID-IJC20&gt;3.0.CO;2-U
    17. Donald J. Buchsbaum. Experimental radioimmunotherapy. Seminars in Radiation Oncology 2000, 10 (2) , 156-167.

    Pair your accounts.

    Export articles to Mendeley

    Get article recommendations from ACS based on references in your Mendeley library.

    Pair your accounts.

    Export articles to Mendeley

    Get article recommendations from ACS based on references in your Mendeley library.

    You’ve supercharged your research process with ACS and Mendeley!

    STEP 1:
    Click to create an ACS ID

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Your Mendeley pairing has expired. Please reconnect