Summary
These studies have shown that formulation development of a monoclonal anti-body radio-immunoscintigraphy agent is a challenging task involving a number of issues related to the radiochemistry of labeling as well as the stability of the antibody. Through a systematic approach, as described in this study, a stable and efficacious product of high quality can be developed in a rational and efficient manner. In developing an optimized formulation of Tc-99m—antifibrin Fab’ for use in immuno-scintigraphy of DVT, a number of critical components were examined in regard to technetium-labeling chemistry and the ability of the formulation to support the long-term stability of the product. It was found that the addition of glucarate as a transfer ligand, SnCl2 as a reducing agent, and neutral-to-acidic pH of the solution were essential for optimum radiolabeling of 0.5 mg of antifibrin Fab’ to a desired activity of 25 mCi of Tc-99m. The lyophilization of the final product was also required to further stabilize both the antifibrin Fab’ fragment and the reducing agent. The addition of carbohydrate as bulking agent and lyoprotectant and inclusion of EDTA as a chelating agent further improved the performance of the formulations, resulting in products with long shelf-life.
Many of the principles described in this study are not only useful in developing a technetium-based immunoscintigraphic agent but are also applicable to other immunopharmaceuticals, including products involving delivery of radionuclides, drugs, and toxins for immunotherapy of cancer and other diseases.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Bogard, W., Dean, R., Deo, Y., Fuchs, R., Mattis, J., McLean, A., and Berger, H., 1989, Practical considerations in the production, purification, and formulation of monoclonal antibodies for immunoscintigraphy and immunotherapy, Semin. Nucl. Med. 19:202–220.
Eckelman, W. C., Paik, C. H., and Steigman, J., 1989, Three approaches to radiolabeling antibodies with 99mTc, Nucl. Med. Biol. 16:171–176.
Fischman, A., Rubin, R., Khaw, B. A., Krammer, P. N., Wilkinson, R., Ahamad, M., Nedelman, M., Locke, E., Nossiff, N.D., and Strauss, W. H., 1989, Radionuclide imaging of experimental atherosclerosis with non-specific polyclonal immunoglobulin G, J. Nucl. Med. 30:1095–1100.
Goldenberg, D., Wlodkowski, T, Sharkey, R., Silberstein, E., Serafini, A., Garty, I., Van Heertum, R. L., Higginbolham-ford, E., Kotler, J., Balasubramaniam, N., Swayne, L., Hansen, H., and Pinsky, C., 1993, Colorectal cancer imaging with iodine-123-labeled CEA monoclonal antibody fragments, J. Nucl. Med. 34:61–70.
Hnatowich, D. J., 1991, Recent developments in the radiolabeling of antibodies with iodine, indium, and technetium, Semin. Nucl. Med. 20:80–91.
Khaw, B. A., Strauss, W. A., Moore, R., Fallon, J., Yasuda, T., Gold, H., and Haber, E., 1987, Myocardial damage delineated by indium-111 antimyosin Fab and technetium-99m pyrophosphate, J. Nucl. Med. 28:76–82.
Knight, I., Maurer, A., Ammar, I., Shealy, D., and Mattis, J., 1988, Evaluation of indium-lll-labeled antifibrin antibody for imaging vascular thrombi, J. Nucl. Med. 29:494–502.
Kudryk, B., Rohoza, A., and Ahadi, M., 1983, A monoclonal antibody with ability to distinguish between NH2-terminal fragments derived from fibrinogen and fibrin, Mol. Immunol. 20:1191–1200.
Kudryk, B., Rohoza, A., and Ahadi, M., 1984, Specificity of a monoclonal antibody for the NH-2 terminal region of fibrin, Mol. Immunol. 21:89–94.
Paik, C. H., Eckelnian, W., and Reba, R., 1986, Transchelation of 99mTc from low affinity sites to high affinity sites of antibody, Nucl. Med. Biol. 13:369–362.
Pak, K. Y., Nedelman, M. A., Fogler, W. E., Tarn, S. H., Wilson, E., Van Haarlem, L. J. M., Colognola, R., Warnaar, S. A., and Daddona, P. E., 1991, Evaluation of the 323/A3 monoclonal antibody and the use of technetium-labeled 323/A3 Fab’ for the detection of Pan adenocarcinoma, Nucl. Med. Biol. 18:483–497.
Pak, K. Y, Nedelman, M., Kanke, M., Khaw, B. A., Mattis, J. A., Strauss, W. H., Dean, R., and Berger, H. J., 1992, An instant kit method for labeling antimyosin Fab’ with technetium-99m: Evaluation in an experimental myocardial infarct model, J. Nucl. Med. 33:144–150.
Pietersz, G. A., 1990, The linkage of cytotoxic drugs to monoclonal antibodies for the treatment of canter, Bioconj. Chem. 1:89–95.
Rhodes, B., 1991, Direct labeling of proteins with 99m-Tc, Nucl. Med. Biol. 18: 667–676.
Robyt, J. F, Ackerman, R. J., and Chitrenden, C. G., 1971, Reaction of protein disulfide groups with Ell-man’s reagent: a case study of the number of sulfhydryl and disulfide groups in asporgillus oryzaedamylase, papaine, and lysozyme, Arch. Biochem. Biophys. 147:262–269.
Rosebrough, S., McAfee, J., Grossman, Z., and Schemancik, L., 1989, Immunoreactivity of 111-In and 131-I fibrin specific monoclonal antibody used for thrombus imaging, J. Immunol. Methods 116:123–129.
Rubin, R., Fischman, A. J., Callahan, R., Khaw, B. A., Keech, F., Ahamad, M., Wilkinson, R., and Strauss, W. H., 1989, 111 In-labeled nonspecific immunoglobulin scanning in the detection of focal infection, N. Engl. J. Med. 321:935–940.
Schaible, T., and Alavi, A., 1991, Antifibrin scintigraphy in the diagnostic evaluation of acute deep venous thrombosis, Semin. Nucl. Med. 21:313–324.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2002 Kluwer Academic Publishers
About this chapter
Cite this chapter
Kamat, M.S., Tolman, G.L., Brown, J.M. (2002). Formulation Development of an Antifibrin Monoclonal Antibody Radiopharmaceutical. In: Pearlman, R., Wang, Y.J. (eds) Formulation, Characterization, and Stability of Protein Drugs: Case Histories. Pharmaceutical Biotechnology, vol 9. Springer, Boston, MA. https://doi.org/10.1007/0-306-47452-2_9
Download citation
DOI: https://doi.org/10.1007/0-306-47452-2_9
Publisher Name: Springer, Boston, MA
Print ISBN: 978-0-306-45332-8
Online ISBN: 978-0-306-47452-1
eBook Packages: Springer Book Archive