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.

MENDELEY PAIRING EXPIRED
Your Mendeley pairing has expired. Please reconnect
ACS Publications. Most Trusted. Most Cited. Most Read
My Activity
CONTENT TYPES

Figure 1Loading Img

Preparation and Characterization of Anti-Tenascin Monoclonal Antibody−Streptavidin Conjugates for Pretargeting Applications

View Author Information
Departments of Radiology and Pathology, Duke University Medical Center, DUMC 3808, Durham, North Carolina 27710
Cite this: Bioconjugate Chem. 1999, 10, 5, 867–876
Publication Date (Web):August 3, 1999
https://doi.org/10.1021/bc990040w
Copyright © 1999 American Chemical Society

    Article Views

    382

    Altmetric

    -

    Citations

    LEARN ABOUT THESE METRICS
    Other access options

    Abstract

    Radioimmunopretargeting is based on the separate injection of a modified mAb and the radionuclide and most frequently exploits the very high avidity of biotin for streptavidin (SA). Currently, we are evaluating the therapeutic potential of directly labeled monoclonal antibody (mAb) 81C6, reactive with the extracellular matrix protein tenascin, in surgically created glioma resection cavity patients. To be able to investigate pretargeting in this setting, the synthesis of 81C6 mAb−SA conjugates was required. In the current study, we have evaluated five methods for preparing both murine 81C6 (m81C6) and human/mouse chimeric 81C6 (c81C6) SA conjugates with regard to yield, biotin-binding capacity, immunoreactivity, and molecular weight. The 81C6 mAb and SA were coupled by covalent interaction between sulfhydryl groups generated on the mAb via N-succinimidyl-S-acetylthioacetate, dithiothreitol or 2-iminothiolane (2IT), and maleimido-derivatized SA, prepared via sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC) or N-succinimidyl-3-(2-pyridyldithio)-propionate. A noncovalent approach involving reaction of a biotinylated mAb, prepared using biotin caproate, and SA also was studied. The evaluation criteria were yield of mAb−SA 215 kDa monomer, as well as conjugate biotin-binding capacity and immunoreactive fraction. The optimal procedure involved activation of m81C6 or c81C6 with 30 equiv of 2IT and reaction of SA with 10 equiv of SMCC and yielded a conjugate with excellent biotin-binding capacity and immunoreactivity. The (125I-labeled m81C6)-2IT-SMCC-SA was stable and did not lose biotin-binding capacity after a 72 h incubation in human glioma cyst fluid in vitro. Although the conjugate was stable in murine serum in vivo, its biotin-binding capacity declined rapidly, consistent with high endogenous biotin levels in the mouse. After injection of the radioiodinated conjugate into athymic mice with subcutaneous D-54 MG human glioma xenografts, high tumor uptake (36.0 ± 10.7% ID/g at 3 days) and excellent tumor:normal tissue ratios were observed.

    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.

    Recommended

    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.

    *

     To whom correspondence should be addressed. Phone:  (919) 684-7705. Fax:  (919) 684-7122. E-mail:  [email protected].

    Cited By

    This article is cited by 8 publications.

    1. Huma Mohsin, Fang Jia, Jeffrey N. Bryan, Geethapriya Sivaguru, Cathy S. Cutler, Alan R. Ketring, William H. Miller, Jim Simón, R. Keith Frank, Louis J. Theodore, Don B. Axworthy, Silvia S. Jurisson, and Michael R. Lewis . Comparison of Pretargeted and Conventional CC49 Radioimmunotherapy Using 149Pm, 166Ho, and 177Lu. Bioconjugate Chemistry 2011, 22 (12) , 2444-2452. https://doi.org/10.1021/bc200258x
    2. Wujun Chen, Yudong Wu, Jie Wang, Wanpeng Yu, Xin Shen, Kai Zhao, Bing Liang, Xiaokun Hu, Shuai Wang, Hongfei Jiang, Xinlin Liu, Miao Zhang, Xiaohui Xing, Chao Wang, Dongming Xing. Clinical advances in TNC delivery vectors and their conjugate agents. Pharmacology & Therapeutics 2024, 253 , 108577. https://doi.org/10.1016/j.pharmthera.2023.108577
    3. Michael R. Lewis, Cathy S. Cutler, Silvia S. Jurisson. Targeted Antibodies and Peptides. 2021, 531-546. https://doi.org/10.1016/B978-0-12-816386-3.00022-3
    4. Peilan Xu, Mingyuan Zou, Shengyu Wang, Li Wang, Lanlan Wang, Fanghong Luo, Ting Wu, Jianghua Yan. Preparation of truncated tissue factor antineuropilin-1 monoclonal antibody conjugate and identification of its selective thrombosis in tumor blood vessels. Anti-Cancer Drugs 2019, 30 (5) , 441-450. https://doi.org/10.1097/CAD.0000000000000767
    5. Igor J. Barani, David A. Larson. Radiation Therapy of Glioblastoma. 2015, 49-73. https://doi.org/10.1007/978-3-319-12048-5_4
    6. Sofia HL Frost, Holger Jensen, Sture Lindegren. In vitro evaluation of avidin antibody pretargeting using 211 At‐labeled and biotinylated poly‐L‐lysine as effector molecule. Cancer 2010, 116 (S4) , 1101-1110. https://doi.org/10.1002/cncr.24798
    7. Fang Jia, Tiffani D. Shelton, Michael R. Lewis. Preparation, Characterization, and Biological Evaluation of a Streptavidin-Chimeric T84.66 Conjugate for Antibody Pretargeting. Cancer Biotherapy and Radiopharmaceuticals 2007, 22 (5) , 654-664. https://doi.org/10.1089/cbr.2007.343-S
    8. Michael R. Lewis, Jiuli Zhang, Fang Jia, Nellie K. Owen, Cathy S. Cutler, Mary F. Embree, Jody Schultz, Louis J. Theodore, Alan R. Ketring, Silvia S. Jurisson, Donald B. Axworthy. Biological comparison of 149Pm-, 166Ho-, and 177Lu-DOTA-biotin pretargeted by CC49 scFv-streptavidin fusion protein in xenograft-bearing nude mice. Nuclear Medicine and Biology 2004, 31 (2) , 213-223. https://doi.org/10.1016/j.nucmedbio.2003.08.004