Preparation Method of (DOTA)n-[Dendrimer PAMAM]- (Trastuzumab)m Conjugate as Radiopharmacutical Compound for Radioimmunotherapy of (177Lu-DOTA)n-[Dendrimer PAMAM]-(Trastuzumab)m
Radioimmunotherapy is one of modalities means of treatment for cancer that exploits the antibody monoclonal specificity to bind to its receptor and the ability of alfa or beta radiation in destroying cancer cells. Trastuzumab is humanized IgG1 monoclonal antibodies which selectively bind to extracellular domain of Human Epidermal Growth Factor Receptor-2 (HER-2). HER-2 has now became a target receptor for treatment of human breast cancer due to their overexpression in the human breast cancer cell surface. This study was aimed in developing a method for preparation of (DOTA) n-[Dendrimer PAMAM]-(trastuzumabm conjugate, a precursor of (177Lu-DOTA)n-PAMAM-(trastuzumab) m radioimmunoconjugate, which is expected to be potential for breast cancer radioimmunotherapy. The conjugate was prepared through several conjugation steps. Characterization of (DOTA)n-[Dendrimer PAMAM]-(trastuzumab)m which was carried out by using a HPLC equipped size exclusion column (SEC) gave a clean peak with retention time (tR) of 10.53 mins which was slower compared to tR of its precursor unconjugated trastuzumab (10.70 mins). This result indicated that molecular weight of (DOTA)n-[Dendrimer PAMAM]-(trastuzumab)m was bigger than its precursor (unconjugated trastuzumab) and pure. The purity of (DOTA)n-[Dendrimer PAMAM]- (trastuzumab)m was verified by measuring the Rf of 177Lu-radiolabeled-(DOTA)n-[Dendrimer PAMAM]-(trastuzumab)m using instance thin layer chromatography-silica gel (ITLC-SG). The ITLC-SG of (DOTA)n-[Dendrimer PAMAM]-(trastuzumab)m which was radiolabeled with 177Lu- gave a clean peak with Rf of <0.3, while the precursors which were regards as impurities, (free 177Lu in form 177Lu-EDTA, 177Lu-DOTA, and 177Lu- dendrimer PAMAM which was incubated as non specific binding), all of them gave Rf of >0.6. These ITLC results indicated that (DOTA)n-[Dendrimer PAMAM]-(trastuzumab)m radiolabeled with 177Lu was pure.
2. Suzanna E, Sirait T, Rahayu PS, Shalmont G, Anwar E, Andalusia R, Harjati, et al. Registrasi kanker berbasis rumah sakit kanker “Dharmais”-Pusat Kanker Nasional, 1993-2007. Ind J of Cancer. 2012.6(4):181- 205.
3. DCE Ng, MBBS, MRCP, FAMS. Radioimunotherapy: A brief review. Biomed Imaging Interv J. 2005.2(3):e23.
4. Rasaneh S, Rajabi H, Babaei MH, Daha FJ. 177Lu labeling of herceptin and preclinical validation as a new radiopharmaceutical for radioimunotherapy of breast cancer. Nucl Med and Biol. 2010.37:949-55.
5. Schrohl A, Pedersen HC, Jensen SS, Nielsen SL, Brünner N. Human epidermal growth factor receptor-2 (HER-2) immunoreactivity: Specificity of three pharmacodiagnostic antibodies. Histopathology. 2011.59(5):975-83.
6. Hermanto S, Haryuni RD, Ramli M, Mutalib A, & Hudiyono S. Preparation of F(ab’)2 trastuzumab fragment for radioimmunoconjugate synthesis of 177Lu-DOTA-F(ab’)2-trastuzumab. IOSR J Pharm. 2012.2(6):12-8.
7. Hermanson GT. Bioconjugate technique. 2nd Ed. London: Elsevier; 2008. 67-9, 283-4.
8. Humani T, Ramli M, Rustendi CT Subur M. Preparasi dan uji stabilitas 177Lu-DOTA-Nimotuzumab sebagai radiofarmaka terapi kanker. 2010. Prosiding Seminar Nasional VI SDM Teknologi Nuklir, Yogyakarta 29 November, 2010: 663-70.
9. Kobayashi H, Sato N, Saga T, Nakamoto Y, Ishimori T, Toyama S, et al. Monoclonal antibody-dendrimer conjugates enable radiolabelling of antibody with markedly high specific activity with minimal loss of imunoreactivity. Eur J Nucl Med. 2000.27(9):1334-9
10. Anonim. Lutetium-177.diambil dari. http:// shop.perkinelmer.com/content/manuals/gde_ lutetium177features.pdf.diakses 14 Februari, 2013 11. Wang AZ, Gu F, Zhang L, Chan JM, Radovic-Moreno A, Shaikh MR, et al. Biofunctionalized targeted nanoparticles for therapeutic applications. Expert Opin. Biol. Ther. 2008.8(8):1063-70.
12. Menjoge AR, Kannan RM, Tomalia DA. Dendrimer- based drug and imaging conjugates: Design considerations for nanomedical applications. Drug Discovery Today. 2010.15:171-84.
13. Zhang Z, Rong F, Niu S, Xie Y, Wang Y, Yang H, et al. Investigation the effects of nano golds on the fluorescence properties of the sectorial poly(amidoamine) (PAMAM) dendrimers. Appl Surf Sci J. 2010.256:7194-9.
14. Bhalgat MK and Roberts JC. Molecular modelling of PAMAM StarburstTM dendrimer. Eur Polym J. 2000.36:647-51.
15. Ramli M, Hidayat B, Ardiyatno CN, Aguswarini S, Karyadi, et al. Preclinical study of 177Lu-DOTA- trastuzumab, a potential radiopharmaceutical for therapy of breast cancer positive HER-2. Proceedings of the 2nd International Seminar on Chemistry 2011, Jatinangor 24-25 November, 2011: 71-8.
16. Anonim. Got Protein?™ kit catalog number 166- 2900EDU testing protein content of common foods using the Quick Start™ Bradford. 2013; [1 tayangan]. diambil dari: URL:http://explorer.bio-rad.com. diakses 5 Februari, 2013.
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