Radiolabeling of Herceptin with 99mTc as a Her2 tracer
Introduction: Trastuzumab is a monoclonal antibody that is used in treating breast cancer. We labeled this monoclonal antibody with Technetium-99m and performed in vitro and in vivo quality control tests as a first step in the production of a new radiopharmaceutical.
Methods: Trastuzumab was labeled with Technetium-99m using Succinimidyl Hydrazinonicotinamide (HYNIC) as chelator. Radiochemical Purity and stability in buffer and serum were determined. Immunoreactivity and toxicity of the complex were tested on SKBR3, MCF7 and A431 breast cancer cell lines. Biodistribution study was performed in normal mice at 4 and 24 h post injection.
Results: The radiochemical purity of the complex was 95±1.4%. The stabilities in phosphate buffer and in human blood serum at 24 h post preparation were 85±3.5% and 74±1.2%, respectively. The immunoreactivity of the complex was 86±1.4%. The binding of labeled antibody to the surface of SKBR3, MCF7 and A431 cells were increased by increasing Her2 concentration on the cells surface.Conclusions: The findings showed that the new radiopharmaceutical can be a promising candidate as Her2 antigen scanning for human breast cancer.
Metro G, Mottolese M, Fabi A. HER-2-positive metastatic breast cancer: trastuzumab and beyond. Expert Opin Pharmacother. 2008 Oct;9(15):2583-601.
Bange J, Zwick E, Ullrich A. Molecular targets for breast cancer therapy and prevention. Nat Med 2001 May;7(5):548–52.
Agrawal A, Gutteridge E, Gee J, Nicholson R, Robertson JR. Overview of tyrosine kinase inhibitors in clinical breast cancer. Endocr Relat Cancer 2005;12: 135-44.
Nahta R, Esteva F. HER-2-Targeted Therapy – Lessons Learned and Future Directions. Clin Cancer Res. 2003 Nov;9(14):5078 – 48.
Albanell J, Codony J, Rovira A, Mellado B, Gascon P. Mechanism of action of anti-HER2 monoclonal antibodies: scientific update on trastuzumab and 2C4. Adv Exp Med Bio. 2003;532:253–68.
Abrams MJ, Juweid M, Tenkate CI. Technetium-99m human polyclonal IgG radiolabeled via the hydrazino nicotinamide derivative for imaging focal sites of infection in rats. J Nucl Med 1990 Dec;31(12):2022–8.
Larsen SK, Solomon HF, Caldwell G, Abrams MJ. [99mTc]tricine: a useful precursor complex for the radiolabeling of hydrazinonicotinate protein conjugates. Bioconjug Chem. 1995 Sep-Oct;6(5):635-8.
King TP, Zhao SW, Lam T. Preparation of protein conjugates via intra molecular hydrazone linkage. Biochem. 1986 Sep;25(19):5774–9.
Lindmo T, Boven E, Cuttitta F, Fedorko J, and Bunn PJ. Determination of the immunoreactive fraction of radiolabeled monoclonal antibodies by linear extrapolation to binding at infinite antigen excess. J Immunol Methods 1984 Aug;72(1):77-89.
Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983 Dec;65(1-2):55-63.
Menard S, Pupa S, Campiglio M, Tagliabue E. Biologic and therapeutic role of HER2 in cancer. Oncogene 2003 Sep;22(42):6570-8.
Tan A and Swain S. Ongoing adjuvant trials with trastuzumab in breast cancer. Semi Oncol 2003 Oct;30(5 Suppl 16):54-64.
Seidman A, Hudis C, Pierri M, Shak S, Paton V, Ashby M, Stewart SJ, Keefe D. Cardiac dysfunction in the trastuzumab clinical trials experience. J Clin Oncol 2002 Mar;20(5):1215-21.
Salouti M, Rajabi H, Babaei H, Rasaee MJ, Najafi R, Paknejad M, Mohammad Hassan Z, Altarihi TM. 99mTc direct radiolabeling of PR81, a new anti-MUC1 monoclonal antibody for radioimmunoscintigraphy. Iran J Nucl Med 2005;13(1):7–16.
Salouti M, Rajabi H, Babaei H, Rasaee MJ, Najafi R, Paknejad M, Mohammad Hassan Z, Altarihi TM. A new monoclonal antibody radiopharmaceutical for radioimmunoscintigraphy of breast cancer: direct labeling of antibody and its quality control. Daru 2006 Mar;14(1):51-6.
Rajabifar S, Akhlaghi M, Jalilian AR, Bolourinovin F, Maashkar B, Talebimehrdar M, Ghafouri M. Preparation and biodistribution of 99mTc-IgG-HYNIC in normal rats, Nukleonika 2009;54(4):279−284.