Abstract
Immunoconjugates—monoclonal antibodies (mAbs) coupled to highly toxic agents, including radioisotopes and toxic drugs (ineffective when administered systemically alone)—are becoming a significant component of anticancer treatments. By combining the exquisite targeting specificity of mAbs with the enhanced tumor-killing power of toxic effector molecules, immunoconjugates permit sensitive discrimination between target and normal tissue, resulting in fewer toxic side effects than most conventional chemotherapeutic drugs. Two radioimmunoconjugates, ibritumomab tiuxetan (Zevalin) and tositumomab-131I (Bexxar), and one drug conjugate, gemtuzumab ozogamicin (Mylotarg), are now on the market. For the next generation of immunoconjugates, advances in protein engineering will permit greater control of mAb targeting, clearance and pharmacokinetics, resulting in significantly improved delivery to tumors of radioisotopes and potent anticancer drugs. Pretargeting strategies, which separate the two functions of antibody-based localization and delivery or generation of the toxic agent into two steps, also promise to afford superior tumor targeting and therapeutic efficacy. Several challenges in optimizing immunoconjugates remain, however, including poor intratumoral mAb uptake, normal tissue conjugate exposure and issues surrounding drug potency and conditional release from mAb carriers. Nonetheless, highly promising results from preclinical models will continue to drive the clinical development of this therapeutic class.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Hoogenboom, H.R. & Chames, P. Natural and designer binding sites made by phage display technology. Immunol. Today 21, 371–378 (2000).
Feldhaus, M.J. & Siegel, R.W. Yeast display of antibody fragments: a discovery and characterization platform. J. Immunol. Methods 290, 69–80 (2004).
Lipovsek, D. & Pluckthun, A. In-vitro protein evolution by ribosome display and mRNA display. J. Immunol. Methods 290, 51–67 (2004).
Irving, R.A., Coia, G., Roberts, A., Nuttall, S.D. & Hudson, P.J. Ribosome display and affinity maturation: from antibodies to single V-domains and steps towards cancer therapeutics. J. Immunol. Methods 248, 31–45 (2001).
Lonberg, N. et al. Antigen-specific human antibodies from mice comprising four distinct genetic modifications. Nature 368, 856–859 (1994).
Green, L.L. et al. Antigen-specific human monoclonal antibodies from mice engineered with human Ig heavy and light chain YACs. Nat. Genet. 7, 13–21 (1994).
Jain, R.K. Tumor physiology and antibody delivery. Front. Radiat. Ther. Oncol. 24, 32–46; discussion 64–68 (1990).
Saga, T. et al. Targeting cancer micrometastases with monoclonal antibodies: a binding-site barrier. Proc. Natl. Acad. Sci. USA 92, 8999–9003 (1995).
Ruiz-Cabello, F., Cabrera, T., Lopez-Nevot, M.A. & Garrido, F. Impaired surface antigen presentation in tumors: implications for T cell-based immunotherapy. Semin. Cancer Biol. 12, 15–24 (2002).
Wahl, R.L. Tositumomab and (131)I therapy in non-Hodgkin's lymphoma. J. Nucl. Med. 46 (Suppl. 1), 128S–140S (2005).
Borghaei, H. & Schilder, R.J. Safety and efficacy of radioimmunotherapy with yttrium 90 ibritumomab tiuxetan (Zevalin). Semin. Nucl. Med. 34, 4–9 (2004).
Linenberger, M.L. CD33-directed therapy with gemtuzumab ozogamicin in acute myeloid leukemia: progress in understanding cytotoxicity and potential mechanisms of drug resistance. Leukemia 19, 176–182 (2005).
Milenic, D.E., Brady, E.D. & Brechbiel, M.W. Antibody-targeted radiation cancer therapy. Nat. Rev. Drug Discov. 3, 488–499 (2004).
Wu, A.M. & Yazaki, P.J. Designer genes: recombinant antibody fragments for biological imaging. Q. J. Nucl. Med. 44, 268–283 (2000).
Hulahov, A. & Chester, K.A. Engineered single chain antibody fragments for radioimmunotherapy. Q. J. Nucl. Med. Mol. Imaging 48, 279–288 (2004).
Adams, G.P. et al. High affinity restricts the localization and tumor penetration of single-chain Fv antibody molecules. Cancer Res. 61, 4750–4755 (2001).
Yazaki, P.J. et al. Tumor targeting of radiometal labeled anti-CEA recombinant T84.66 diabody and T84.66 minibody: comparison to radioiodinated fragments. Bioconjug. Chem. 12, 220–228 (2001).
Borsi, L. et al. Selective targeting of tumoral vasculature: comparison of different formats of an antibody (L19) to the ED-B domain of fibronectin. Int. J. Cancer 102, 75–85 (2002).
Slavin-Chiorini, D.C. et al. Biological properties of chimeric domain-deleted anticarcinoma immunoglobulins. Cancer Res. (Suppl.) 55, 5957s–5967s (1995).
Kenanova, V. et al. Tailoring the pharmacokinetics and positron emission tomography imaging properties of anti-carcinoembryonic antigen single-chain Fv-Fc antibody fragments. Cancer Res. 65, 622–631 (2005).
Albrecht, H. et al. Production of soluble ScFvs with C-terminal-free thiol for site-specific conjugation or stable dimeric ScFvs on demand. Bioconjug. Chem. 15, 16–26 (2004).
Natarajan, A., Xiong, C.Y., Albrecht, H., DeNardo, G.L. & DeNardo, S.J. Characterization of site-specific ScFv PEGylation for tumor-targeting pharmaceuticals. Bioconjug. Chem. 16, 113–121 (2005).
Li, L. et al. Reduction of kidney uptake in radiometal labeled peptide linkers conjugated to recombinant antibody fragments. Site-specific conjugation of DOTA-peptides to a Cys-diabody. Bioconjug. Chem. 13, 985–995 (2002).
Olafsen, T. et al. Covalent disulfide-linked anti-CEA diabody allows site-specific conjugation and radiolabeling for tumor targeting applications. Protein Eng. Des. Sel. 17, 21–27 (2004).
Waibel, R. et al. Stable one-step technetium-99m labeling of His-tagged recombinant proteins with a novel Tc(I)-carbonyl complex. Nat. Biotechnol. 17, 897–901 (1999).
Corneillie, T.M., Whetstone, P.A., Lee, K.C., Wong, J.P. & Meares, C.F. Converting weak binders into infinite binders. Bioconjug. Chem. 15, 1389–1391 (2004).
Sundaresan, G. et al. (124)I-labeled engineered anti-CEA minibodies and diabodies allow high-contrast, antigen-specific small-animal PET imaging of xenografts in athymic mice. J. Nucl. Med. 44, 1962–1969 (2003).
Robinson, M.K. et al. Quantitative immuno-positron emission tomography imaging of HER2-positive tumor xenografts with an iodine-124 labeled anti-HER2 diabody. Cancer Res. 65, 1471–1478 (2005).
Olafsen, T. et al. Characterization of engineered anti-p185HER-2 (scFv-CH3)2 antibody fragments (minibodies) for tumor targeting. Protein Eng. Des. Sel. 17, 315–323 (2004).
Olafsen, T. et al. Optimizing radiolabeled engineered anti-p185HER2 antibody fragments for in vivo imaging. Cancer Res. 65, 5907–5916 (2005).
Smith-Jones, P.M. et al. Imaging the pharmacodynamics of HER2 degradation in response to Hsp90 inhibitors. Nat. Biotechnol. 22, 701–706 (2004).
Begent, R.H.J. et al. Clinical evidence of efficient tumor targeting based on single-chain Fv antibody selected from a combinatorial library. Nat. Med. 2, 979–984 (1996).
Larson, S.M. et al. Single chain antigen binding protein (sFv CC49)—First human studies in colorectal carcinoma metastatic to liver. Cancer 80 (Suppl.), 2458–2468 (1997).
Santimaria, M. et al. Immunoscintigraphic detection of the ED-B domain of fibronectin, a marker of angiogenesis, in patients with cancer. Clin. Cancer Res. 9, 571–579 (2003).
Wong, J.Y. et al. Pilot trial evaluating an 123I-labeled 80-kilodalton engineered anticarcinoembryonic antigen antibody fragment (cT84.66 minibody) in patients with colorectal cancer. Clin. Cancer Res. 10, 5014–5021 (2004).
Adams, G. et al. Delivery of the α-emitting radioisotope bismuth-213 to solid tumors via single-chain Fv and diabody molecules. Nucl. Med. Biol. 27, 339–346 (2000).
Adams, G.P. et al. A single treatment of yttrium-90-labeled CHX-A''-C6.5 diabody inhibits the growth of established human tumor xenografts in immunodeficient mice. Cancer Res. 64, 6200–6206 (2004).
Sharkey, R.M. & Goldenberg, D.M. Perspectives on cancer therapy with radiolabeled monoclonal antibodies. J. Nucl. Med. 46 (Suppl. 1), 115S–127S (2005).
Jhanwar, Y.S. & Divgi, C. Current status of therapy of solid tumors. J. Nucl. Med. 46 (Suppl. 1), 141S–150S (2005).
Wong, J.Y. et al. A phase I trial of 90Y-anti-carcinoembryonic antigen chimeric T84.66 radioimmunotherapy with 5-fluorouracil in patients with metastatic colorectal cancer. Clin. Cancer Res. 9, 5842–5852 (2003).
Sharkey, R.M. et al. A phase I trial combining high-dose 90Y-labeled humanized anti-CEA monoclonal antibody with doxorubicin and peripheral blood stem cell rescue in advanced medullary thyroid cancer. J. Nucl. Med. 46, 620–633 (2005).
Dubowchik, G.M. & Walker, M.A. Receptor-mediated and enzyme-dependent targeting of cytotoxic anticancer drugs. Pharmacol. Ther. 83, 67–123 (1999).
Payne, G. Progress in immunoconjugate cancer therapeutics. Cancer Cell 3, 207–212 (2003).
Trail, P.A. et al. Cure of xenografted human carcinomas by BR96-doxorubicin immunoconjugates. Science 261, 212–215 (1993).
Mosure, K.W., Henderson, A.J., Klunk, L.J. & Knipe, J.O. Disposition of conjugate-bound and free doxorubicin in tumor-bearing mice following administration of a BR96-doxorubicin immunoconjugate (BMS 182248). Cancer Chemother. Pharmacol. 40, 251–258 (1997).
Saleh, M.N. et al. Phase I trial of the anti-Lewis Y drug immunoconjugate BR96-doxorubicin in patients with Lewis Y-expressing epithelial tumors. J. Clin. Oncol. 18, 2282–2292 (2000).
Tolcher, A.W. et al. Cantuzumab mertansine, a maytansinoid immunoconjugate directed to the CanAg antigen: a phase I, pharmacokinetic, and biologic correlative study. J. Clin. Oncol. 21, 211–222 (2003).
Griffiths, G.L. et al. Cure of SCID mice bearing human B-lymphoma xenografts by an anti-CD74 antibody-anthracycline drug conjugate. Clin. Cancer Res. 9, 6567–6571 (2003).
Damle, N.K. & Frost, P. Antibody-targeted chemotherapy with immunoconjugates of calicheamicin. Curr. Opin. Pharmacol. 3, 386–390 (2003).
Damle, N.K. Tumour-targeted chemotherapy with immunoconjugates of calicheamicin. Expert Opin. Biol. Ther. 4, 1445–1452 (2004).
Hamann, P.R. et al. An anti-CD33 antibody-calicheamicin conjugate for treatment of acute myeloid leukemia. Choice of linker. Bioconjug. Chem. 13, 40–46 (2002).
Hamann, P.R. et al. Gemtuzumab ozogamicin, a potent and selective anti-CD33 antibody-calicheamicin conjugate for treatment of acute myeloid leukemia. Bioconjug. Chem. 13, 47–58 (2002).
Bross, P.F. et al. Approval summary: gemtuzumab ozogamicin in relapsed acute myeloid leukemia. Clin. Cancer Res. 7, 1490–1496 (2001).
Sievers, E.L. et al. Selective ablation of acute myeloid leukemia using antibody-targeted chemotherapy: a phase I study of an anti-CD33 calicheamicin immunoconjugate. Blood 93, 3678–3684 (1999).
Sievers, E.L. & Linenberger, M. Mylotarg: antibody-targeted chemotherapy comes of age. Curr. Opin. Oncol. 13, 522–527 (2001).
Giles, F.J. et al. Mylotarg (gemtuzumab ozogamicin) therapy is associated with hepatic venoocclusive disease in patients who have not received stem cell transplantation. Cancer 92, 406–413 (2001).
DiJoseph, J.F. et al. Antibody-targeted chemotherapy with CMC-544: a CD22-targeted immunoconjugate of calicheamicin for the treatment of B-lymphoid malignancies. Blood 103, 1807–1814 (2004).
DiJoseph, J.F. et al. Antibody-targeted chemotherapy of B-cell lymphoma using calicheamicin conjugated to murine or humanized antibody against CD22. Cancer Immunol. Immunother. 54, 11–24 (2005).
Boghaert, E.R. et al. Antibody-targeted chemotherapy with the calicheamicin conjugate hu3S193-N-acetyl gamma calicheamicin dimethyl hydrazide targets Lewis(y) and eliminates Lewis(y-positive) human carcinoma cells and xenografts. Clin. Cancer Res. 10, 4538–4549 (2004).
Linenberger, M.L. et al. Multidrug-resistance phenotype and clinical responses to gemtuzumab ozogamicin. Blood 98, 988–994 (2001).
Hamann, P.R. et al. An anti-MUC1 antibody-calicheamicin conjugate for treatment of solid tumors. Choice of linker and overcoming drug resistance. Bioconjug. Chem. 16, 346–353 (2005).
Vitetta, E.S., Thorpe, P.E. & Uhr, J.W. Immunotoxins: magic bullets or misguided missiles? Immunol. Today 14, 252–259 (1993).
Thorpe, P.E. et al. Improved antitumor effects of immunotoxins prepared with deglycosylated ricin A-chain and hindered disulfide linkages. Cancer Res. 48, 6396–6403 (1988).
Smith, S.V. Technology evaluation: cantuzumab mertansine, ImmunoGen. Curr. Opin. Mol. Ther. 6, 666–674 (2004).
Xie, H., Audette, C., Hoffee, M., Lambert, J.M. & Blattler, W.A. Pharmacokinetics and biodistribution of the antitumor immunoconjugate, cantuzumab mertansine (huC242–DM1), and its two components in mice. J. Pharmacol. Exp. Ther. 308, 1073–1082 (2004).
Lutz, R.L. et al. HuC242–DM4, an antibody-maytansinoid conjugate with superior preclinical activity in human CanAg-positive tumor xenograft models in SCID mice. Abstract of a paper presented at the 96th annual meeting of the American Association for Cancer Research, Anaheim, CA, 16–20 April 2005.
Fosella, F.V. et al. Phase II trial of BB-10901 (huN901–DM1) given weekly for four consecutive weeks every 6 weeks in patients with relapsed SCLC and CD56-positive small cell carcinoma. Abstract of a paper presented at the 41st annual meeting of the American Society of Clinical Oncology, Orlando, FL, 13–17 May 2005.
Tassone, P. et al. Cytotoxic activity of the maytansinoid immunoconjugate B-B4–DM1 against CD138+ multiple myeloma cells. Blood 104, 3688–3696 (2004).
Henry, M.D. et al. A prostate-specific membrane antigen-targeted monoclonal antibody-chemotherapeutic conjugate designed for the treatment of prostate cancer. Cancer Res. 64, 7995–8001 (2004).
Dubowchik, G.M. & Firestone, R.A. Cathepsin B-sensitive dipeptide prodrugs. 1. A model study of structural requirements for efficient release of doxorubicin. Bioorg. Med. Chem. Lett. 8, 3341–3346 (1998).
Dubowchik, G.M., Mosure, K., Knipe, J.O. & Firestone, R.A. Cathepsin B-sensitive dipeptide prodrugs. 2. Models of anticancer drugs paclitaxel (Taxol), mitomycin C and doxorubicin. Bioorg. Med. Chem. Lett. 8, 3347–3352 (1998).
Dubowchik, G.M. et al. Cathepsin B-labile dipeptide linkers for lysosomal release of doxorubicin from internalizing immunoconjugates: model studies of enzymatic drug release and antigen-specific in vitro anticancer activity. Bioconjug. Chem. 13, 855–869 (2002).
Walker, M.A., Dubowchik, G.M., Hofstead, S.J., Trail, P.A. & Firestone, R.A. Synthesis of an immunoconjugate of camptothecin. Bioorg. Med. Chem. Lett. 12, 217–219 (2002).
Walker, M.A. et al. Monoclonal antibody mediated intracellular targeting of tallysomycin S(10b). Bioorg. Med. Chem. Lett. 14, 4323–4327 (2004).
Doronina, S.O. et al. Development of potent monoclonal antibody auristatin conjugates for cancer therapy. Nat. Biotechnol. 21, 778–784 (2003).
Francisco, J.A. et al. cAC10-vcMMAE, an anti-CD30-monomethyl auristatin E conjugate with potent and selective antitumor activity. Blood 102, 1458–1465 (2003).
Saad, E.D. et al. Phase II study of dolastatin-10 as first-line treatment for advanced colorectal cancer. Am. J. Clin. Oncol. 25, 451–453 (2002).
Hamblett, K.J. et al. Effects of drug loading on the antitumor activity of a monoclonal antibody drug conjugate. Clin. Cancer Res. 10, 7063–7070 (2004).
Law, C.L. et al. Efficient elimination of B-lineage lymphomas by anti-CD20-auristatin conjugates. Clin. Cancer Res. 10, 7842–7851 (2004).
Beeson, C. et al. Conditionally cleavable radioimmunoconjugates: a novel approach for the release of radioisotopes from radioimmunoconjugates. Bioconjug. Chem. 14, 927–933 (2003).
Arano, Y. Strategies to reduce renal radioactivity levels of antibody fragments. Q. J. Nucl. Med. 42, 262–270 (1998).
Studer, M., Kroger, L.A., DeNardo, S.J., Kukis, D.L. & Meares, C.F. Influence of a peptide linker on biodistribution and metabolism of antibody-conjugated benzyl-EDTA. Comparison of enzymatic digestion in vitro and in vivo. Bioconjug. Chem. 3, 424–429 (1992).
Wang, Z. et al. Biotinylation, pharmacokinetics, and extracorporeal adsorption of humanized MAb 111In-MN14 using an avidin-affinity column in rats. Cancer Biother. Radiopharm. 18, 365–375 (2003).
Goodwin, D.A., Meares, C.F., McCall, M.J., McTigue, M. & Chaovapong, W. Pre-targeted immunoscintigraphy of murine tumors with indium-111-labeled bifunctional haptens. J. Nucl. Med. 29, 226–234 (1988).
Sharkey, R.M. et al. Improving the delivery of radionuclides for imaging and therapy of cancer using pretargeting methods. Clin. Cancer Res. (in the press) (2005).
Weiden, P.L. et al. Pretargeted radioimmunotherapy (PRITTM) for treatment of non-Hodgkin's lymphoma (NHL): initial phase I/II study results. Canc. Biother. Radiopharm. 15, 15–29 (2000).
Le Doussal, J.M., Martin, M., Gautherot, E., Delaage, M. & Barbet, J. In vitro and in vivo targeting of radiolabeled monovalent and divalent haptens with dual specificity monoclonal antibody conjugates: enhanced divalent hapten affinity for cell-bound antibody conjugate. J. Nucl. Med. 30, 1358–1366 (1989).
Wang, Y. et al. Pretargeting with amplification using polymeric peptide nucleic acid. Bioconjug. Chem. 12, 807–816 (2001).
He, J. et al. Amplification targeting: a modified pretargeting approach with potential for signal amplification—proof of a concept. J. Nucl. Med. 45, 1087–1095 (2004).
Dübel, S. et al. Bifunctional and multimeric complexes of streptavidin fused to single chain antibodies (scFv). J. Immunol. Methods 178, 201–209 (1995).
Schultz, J. et al. A tetravalent single-chain antibody-streptavidin fusion protein for pretargeted lymphoma therapy. Cancer Res. 60, 6663–6669 (2000).
Goshorn, S. et al. Preclinical evaluation of a humanized NR-LU-10 antibody-streptavidin fusion protein for pretargeted cancer therapy. Cancer Biother. Radiopharm. 16, 109–123 (2001).
Forero, A. et al. Phase 1 trial of a novel anti-CD20 fusion protein in pretargeted radioimmunotherapy for B-cell non-Hodgkin lymphoma. Blood 104, 227–236 (2004).
Rossi, E.A. et al. Pretargeting of CEA-expressing cancers with a trivalent bispecific fusion protein produced in myeloma cells. Clin. Cancer Res. (in the press) (2005).
Senter, P.D. & Springer, C.J. Selective activation of anticancer prodrugs by monoclonal antibody-enzyme conjugates. Adv. Drug Deliv. Rev. 53, 247–264 (2001).
Sharma, S.K., Bagshawe, K.D. & Begent, R.H. Advances in antibody-directed enzyme prodrug therapy. Curr. Opin. Investig. Drugs 6, 611–615 (2005).
Siemers, N.O. et al. Construction, expression, and activities of L49-sFv-beta-lactamase, a single-chain antibody fusion protein for anticancer prodrug activation. Bioconjug. Chem. 8, 510–519 (1997).
Cortez-Retamozo, V. et al. Efficient cancer therapy with a nanobody-based conjugate. Canc. Res. 64, 2853–2857 (2004)
Bhatia, J. et al. Catalytic activity of an in vivo tumor targeted anti-CEA scFv::carboxypeptidase G2 fusion protein. Int. J. Cancer 85, 571–577 (2000).
Francis, R.J. et al. A phase I trial of antibody directed enzyme prodrug therapy (ADEPT) in patients with advanced colorectal carcinoma or other CEA producing tumours. Br. J. Cancer 87, 600–607 (2002).
Mayer, A. et al. Modifying an immunogenic epitope on a therapeutic protein: a step towards an improved system for antibody-directed enzyme prodrug therapy (ADEPT). Br. J. Cancer 90, 2402–2410 (2004).
Torgov, M.Y., Alley, S.C., Cerveny, C.G., Farquhar, D. & Senter, P.D. Generation of an intensely potent anthracycline by a monoclonal antibody-beta-galactosidase conjugate. Bioconjug. Chem. 16, 717–721 (2005).
Acknowledgements
The authors are grateful to their many colleagues and collaborators who have contributed over the years to the fields of antibody engineering and immunoconjugates. Apologies are extended to any authors whose work has not been included as a result of lack of space. Work in Anna Wu's laboratory was supported by NIH CA43904, CA86306, CA92131, DAMD 17-00-1-150 and DAMD 17-00-1-203. Work in Peter Senter's laboratory was partially supported by NIH CA088583.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Rights and permissions
About this article
Cite this article
Wu, A., Senter, P. Arming antibodies: prospects and challenges for immunoconjugates. Nat Biotechnol 23, 1137–1146 (2005). https://doi.org/10.1038/nbt1141
Published:
Issue Date:
DOI: https://doi.org/10.1038/nbt1141
This article is cited by
-
Antibody-drug conjugates in cancer therapy: innovations, challenges, and future directions
Archives of Pharmacal Research (2024)
-
Highlighting the Undetectable — Fluorescence Molecular Imaging in Gastrointestinal Endoscopy
Molecular Imaging and Biology (2023)
-
Imaging in Tumor Immunology
Nuclear Medicine and Molecular Imaging (2021)
-
Assessment of Cellular Uptake Efficiency According to Multiple Inhibitors of Fe3O4-Au Core-Shell Nanoparticles: Possibility to Control Specific Endocytosis in Colorectal Cancer Cells
Nanoscale Research Letters (2020)
-
Development of 177Lu-scFvD2B as a Potential Immunotheranostic Agent for Tumors Overexpressing the Prostate Specific Membrane Antigen
Scientific Reports (2020)