2009 1 1 1 47

Editorial 2 2 In this first editorial for the “first issue” of Avicenna Journal of Medical Biotechnology (AJMB), I thought I should provide our dear readers a little background as well as the reasons used in making decision to publish an English journal in the field of Medical Biotechnology in Iran. Although the field of Medical Biotechnology is still at its infancy in many ways, it has travelled a long journey since its inception in early 1980s, a period parallel to the emergence of DNA recombinant technology and genetic engineering. At that time, the impact of genetic engineering in medicine was predicted to be great and this proved to be correct since the science of Medical Biotechnology expanded later on to include a variety of fields such as molecular medicine, cellular and molecular therapies, applied immunology, nanomedicine, genomics and proteomics, bioinformatics, tissue engineering, biosensors and pharmaceuticals. As a consequence of the great impacts that Medical Biotechnology has had in commercial, legal, ethical and social areas, AJMB is intending to provide a venue for articles discussing these issues in specific areas pertaining to the application of technologies in medicine in general. Since the establishment of biotechnology research centers in Iran, including National Center for Biotechnology and Genetic Engineering more than 15 years ago as well as Avicenna Research Institute, several English language journals have been published in the field of Biotechnology in Iran. AJMB is another English journal in this field but what makes it distinguishable from other journals is its scope of coverage and the comprehensiveness of the fields in medical biotechnology. Regarding the journal’s name, after many hours of deliberations, the scientific staff at Avicenna Research Institute voted for the selection of the name “Avicenna” because of three reasons: 1. Avicenna simply best represents the Iranian-Islamic scientific culture and medicine in history 2. Avicenna is a name that is well recognized and respected in science and medicine throughout the world. 3. Avicenna is a name that has been chosen for a research institute that is focused on the application of biotechnologies in the areas of reproductive biology and cancer. Therefore, it is most appropriate to have Avicenna as the name for a Journal that publishes articles in Medical Biotechnology. As we all know, Avicenna is a Latinized name of Ibn-e-Sina and his contributions to almost all fields of medical sciences and philosophy in the world is well known. Recently, Avicenna’s contributions to science and medicine have been recognized by major international organizations. For example, the United Nations Educational, Scientific and Cultural Organization (UNESCO) has established a virtual campus dedicated to open distance learning in eleven Mediterranean countries under the name of The Avicenna Virtual Campus Project. Also another project with the name of Avicenna Directories of Educational Institutions for the Health Professionals has been initiated under the auspices of the World Health Organization (WHO). I hope the name of “Avicenna” at both Avicenna Research Institute and Avicenna Journal of Medical Biotechnology provides the inspiration for conducting good science and become a place where high quality articles are published. I look forward to receiving your articles in the future and hope the readers of AJMB make this journal, the journal of their choice to publish the results of their research work in the field of Medical Biotechnology. 2 2 Ali M. Ardekani Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Editorial 156.pdf ####

The Place of Avicenna in the History of Medicine 2 2 Avicenna, a Muslim scientist of the tenth and eleventh centuries has an important place in the history of medicine in Iran and the world. Furthermore, the modern medicine is laid upon the infrastructure of his medicine. In this article, the position of Avicenna in the medical history and the scientific influence of his medical works in particular Al-Canon in the development of medical literature and medical educational programs have been studied in a historical approach. In reviewing the position of Avicenna in the history of medicine in the Islamic world and the Europe, it was concluded that during 11th to 17th centuries, the scientific and educational activities of medicine in the world were moving on the pivot of Avicenna medicine or was under its intensive influence. 3 8 Jamal Moosavi Department of History and Civilization of Islamic Nations, University of Tehran Department of History and Civilization of Islamic Nations, University of Tehran Iran AvicennaAvicenna MedicineIslamic MedicineMedical HistoryMedical LiteratureTraditional Medicine 1.pdf Nafisi S. Avicenna in Europe. In: Jashn Nameh Ibn Sina (Vol.2). Tehran: Council of National Works Press;1334 A.H.,300-307. ##Al-Baba M Z. Introduction of Min Moallifat Ibn Sina al-Tibbiya, Al-Maahad al-Torath al-Elmi al-Arabi. Aleppo: Institute of Arabic and Scientific Heritage Press;1984. ##Najmabadi M. History of medicine in Iran, Section 5. Tehran: University of Tehran Press; 1375 A.H., 477-633. ##Azimjan A. A study of Canon of Avicenna in Uzbekistan. In: Jashn Nameh Ibn Sina, Vol.2, pp. 398-402, Council of National Works Press, Tehran, 1334 A.H. ##A bibliography of publication on Islamic and Muslim world since 1906. 8th ed. Brill Academic Publisher; 2006. ##Meshkat SM. Avicenna Angelology. 2nd ed. Hama-dan & Tehran: Bu Ali University Press;1383 A.H. ##Al-Baba MZ. Avicenna Min Moallifat Ibn Sina al-Tibbiya (Kitab daf? al-madarr al-kolya an al-Abdan alensanyya & al-Urjuza fi al-Tibb & al-Adwia alqalbyya). Aleppo: Institute of Arabic and Scientific Heritage Press;1984. ##Elgood CL. A medical history of Persia and the Eastern Caliphate. Cambridge; 1951. ##Ullmann M. Islamic medicine translated into Persian by Fereydoun Badrei. Tehran: Tus Pub-lication; 1383 A.H. ##Rahimov J. About some medical teachings of Avicenna. In: Jashn Nameh Ibn Sina Vol.2. Tehran: Council of National Works Press; 1334 A.H.,112-119. ##Ternovski VN. Influence of medical thought of Avicenna on the Soviet Union physicians. In: Jashn Nameh Ibn Sina, Vol.2. Tehran: Council of National Works Press;1334 A.H.,45-54. ##Tajdbakhsh H. History of veterinary medicine and medicine of Iran (Islamic Era), Vol.2, Chapter 30 & 31, Tehran: University of Tehran Press; 1375 (1996). ##Mostafawi J. Asthma in view point of Avicenna and its comparing with standards of modern medicine. In: Jashn Nameh Ibn Sina, Vol. 2, Tehran: Council of National Works Press;1334 A.H.,91-111. ##Nafisi A. Some biliary herbal drugs which has been mentioned in the second book of Kanun and comparing them with Modern Pharmacology. In: Jashn Nameh Ibn Sina, Vol.2. Tehran: Council of National Works Press;1334 A.H.,55-68. ##Zill al-Rahman HS. Commentators and translators of Kanun, translated into Persian by Abd al-Qadir Hashimi. Tehran: Society for the appreciation of Cultural Works & Dignitaries Press; 1383 (2004). ##Ibn Abi Usaibaa. Uyun al-Anba fi Tabaqat al Atibba, Dar al-Thiqafa. (Vol.3) 4th ed. Beirut: House of Culture Press;1987. ##Brown EG. Arabian medicine. London: 1921. ##Daraie B. Qifti, Tarikh al-Hukama (History of Physicians). Tehran: University of Tehran Press; 1371. ##Samarqandi A. Chahar maqala. Leiden:1909. ##Elgood CL. Safavid medical practice. London: 1970. ##Smith ES. Tibb in the Encyclopedia of Islam, Vol 10. Leiden: Brill Press;2000;452-460. ##Muhaqqeq M. Introduction of the book of Angelology (see: Bib. No. 6). ##Nasr SH. Existential sciences in the history of Iran, Ed. Richard N. Frye. Translated into Persian by Hassan Anoosheh, Vol.4, Section 12. Tehran: Amir Kabir Press;1372 (1993),346-364, ##Ahmadyya A. Method of diagnosis of treatment in view point of Avicenna. In: Jashn Nameh Ibn Sina, Vol.2.Tehran: Council of National Works Press; 1334 A.H., 478-488. ##Hossaini MM. Tuhfa Hakim Momin. Author introduction. 2nd ed. Tehran: Book Store of Mahmoudi Press; 1376(1997). ##Osler SW. Evolution of modern medicine. Oxford: University Press;1923. ##Zajachowski A. The works of Avicenna in Poland. In: Jashn Nameh Ibn Sina, Vol.2.Tehran: Council of National Works Press;1334 A.H.,308-315. ##Mustafa U. Avicenna and medical teachings in Europe in the middle ages. In: Al-Kitab al-Zahabi. Cairo: Golden Book;1952,359-360. ##Sharaf. Ibn Sina. In: Big Encyclopedia of Islam (D.B.A.), Vol.4. Tehran:1370 (1991),1-29. ##

Nutrigenomics and Cancer 2 2 Cancer incidence is projected to increase in the future and an effectual preventive strategy is required to face this challenge. Alteration of dietary habits is potentially an effective approach for reducing cancer risk. Assessment of biological effects of a specific food or bioactive component that is linked to cancer and prediction of individual susceptibility as a function of nutrient-nutrient interactions and genetics is an essential element to evaluate the beneficiaries of dietary interventions. In general, the use of biomarkers to evaluate individuals susceptibilities to cancer must be easily accessible and reliable. However, the response of individuals to bioactive food components depends not only on the effective concentration of the bioactive food components, but also on the target tissues. This fact makes the response of individuals to food components vary from one individual to another. Nutrigenomics focuses on the understanding of interactions between genes and diet in an individual and how the response to bioactive food components is influenced by an individual’s genes. Nutrients have shown to affect gene expression and to induce changes in DNA and protein molecules. Nutrigenomic approaches provide an opportunity to study how gene expression is regulated by nutrients and how nutrition affects gene variations and epigenetic events. Finding the components involved in interactions between genes and diet in an individual can potentially help identify target molecules important in preventing and/or reducing the symptoms of cancer. 9 17 Ali M. Ardekani Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Sepideh Jabbari Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Bioactive food componentsBiomarkerCancer preventionNutrigeneticsNutrigenomics 2.pdf Davis CD, Milner J. Frontiers in nutrigenomics, proteomics, metabolomics and cancer prevention. Mutat Res 2004;551(1-2):51-64. ##Davis CD. Nutritional Interactions: Credentialing of molecular targets for cancer prevention. Exp Biol Med 2007;232(2):176-183. ##Surh YJ. Cancer chemoprevention with dietary phyto-chemicals. Nat Rev Cancer 2003;3(10):768-780. ##Johaning GL, Heimburger DC, Piyathilake CJ. DNA methylation and diet in cancer. J Nutr 2002; 132(12):3814S-3818S. ##McCarty MF. Targeting multiple signaling path-ways as a strategy for managing prostate cancer: multifocal signal modulation therapy. Integr Can-cer Ther 2004;3(4):349-380. ##Hanahan D, Weinberg RA. The hallmarks of can-cer. Cell 2000;100(1):57-70. ##Manson MM. Inhibition of survival signalling by dietary polyphenols and indole-3-carbinol. Eur J Cancer 2005;41(13):1842-1853. ##Suganuma M, Okabe S, Kai Y, Sueoka N, Sueoka E, Fujiki H. Synergistic effects of (epigallocatechin gallate with epicatechin, sulindac, or tamoxifen on cancer preventive activity in the human lung cancer cell line PC-9. Cancer Res 1999;59(1):44-47. ##Boileau TW, Liao Z, Kim S, Lemeshow S, Erdman JW Jr, Clinton SK. Prostate carcinogenesis in N-methyl-N-nitrosourea (NMU)- testosterone- treated rats fed tomato powder, lycopene, or energy restricted diets. J Natl Cancer Inst 2003;95(21): 1578-1586. ##Lu QJ, Huang CY, Yao SX, Wang RS, Wu XN. Effects of fat soluble extracts from vegetable powder and beta carotene on proliferation and apoptosis of lung cancer cell YTMLC-90. Biomed Environ Sci 2003;16(3):237-245. ##Thiagarajan DG, Bennink MR, Bourquin LD, Kavas FA. Prevention of precancerous colonic lesions in rats by soy flakes, soy flour, genistein, and calcium. Am J Clin Nutr 1998;68(Suppl 6): 1394S-1399S. ##Roberts MA, Mutch DM, German JB. Genomics: food and nutrition. Curr Opin Biotechnol 2001;12 (5):516-522. ##Peregrin T. The new frontier of nutrition science: nu-trigenomics. J Am Diet Assoc 2001;101(11):1306. ##Elliott R, Ong TJ. Nutritional genomics. BMJ 2002;324(7351):1438-1442. ##Daniel H. Genomics and proteomics: importance for the future of nutrition research. Br J Nutr 2002; 87(Suppl 2):S305-311. ##van Ommen B, Stierum R. Nutrigenomics: exploi-ting systems biology in the nutrition and health arena. Curr Opin Biotechnol 2002;13(5):517-521. ##Watkins SM, Reifsnyder PR, Pan HJ, German JB, Leiter EH. Lipid metabolome-wide effects of the PPAR-? agonist rosiglitazone. J Lipid Res 2002;43 (11):1809-1817. ##MacBeath G. Protein microarrays and proteomics. Nature Genet 2002;32(Suppl):526-532. ##Grody WW. Molecular genetic risk screening. Annu Rev Med 2003;54:473-490. ##Bailey LB, Gregory JF. Polymorphisms of methyl-enetetrahydrofolate reductase and other enzymes: metabolic significance, risks and impact on folate requirement. J Nutr 1999;129(5):919-922. ##Qi L, Cho YA. Gene environment interaction and obesity. Nutr Rev 2008;66(12):684-694. ##Dahlman I, Vaxillaire M, Nilsson M, Lecoeur C, Gu HF, Cavalcanti Proença C, et al. Estrogen receptor alpha gene variants associate with type 2 diabetes and fasting plasma glucose. Pharmaco-genet Genomics 2008;18(11):967-975. ##Ordovas JM, Shen J. Gene-environment inter actions and susceptibility to metabolic syndrome and other chronic diseases. J Periodontol 2008;79 (Suppl 8):1508-1513. ##Caprioli J, Mele C, Mossali C, Gallizioli L, Giachetti G, Noris M. Polymorphisms of EDNRB, ATG, and ACE genes in salt-sensitive hyper-tension. Can J Physiol Pharmacol 2008;86:(8): 505-510. ##Omer RE, Verhoef L, Van't Veer P, Idris MO, Kadaru AM, Kampman E, et al.. Peanut butter intake, GSTM1 genotype and hepatocellular carci-noma: a case-control study in Sudan. Cancer Causes Control 2001;12(1):23-32. ##Sachidanandam R, Weissman D, Schmidt SC, Kakol JM, Stein LD, Marth G, et al. A map of human genome sequence variation containing 1.42 million single nucleotide polymorphisms. Nature 2001;409(6822):928-933. ##Potter JD. At the interfaces of epidemiology, genetics and genomics. Nat Rev Genet 2001;2(2): 142-147. ##Kong A, Gudbjartsson DF, Sainz J, Jonsdottir GM, Gudjonsson SA, Richardsson B, et al. A high resolution recombination map of the human genome. Nat Genet 2002;31(3):241-247. ##Boomsma D, Busjahn A, Peltonen L. Classical twin studies and beyond. Nat Rev Genet 2002;3 (11):872-882. ##Davis CD, Milner JA. Diet and cancer prevention. In: Temple NJ, Wilson T, Jacobs DV, (eds). Nutritional Health: Strategies for disease preven-tion. Totowa: Humana Press; 2006,151-171. ##Surh YJ. Cancer chemoprevention with dietary phytochemicals. Nat Rev Cancer 2003;3(10):768-780. ##Zhou C, Verma S, Blumberg B. The steroid and xenobiotic receptor (SXR), beyond xenobiotic metabolism. Nucl Recept Signal 2009;7:1-21. ##Takeshita A, Koibuchi N, Oka J, Taguchi M, Shishiba Y, Ozawa Y. Bisphenol-A, an environ-mental estrogen, activates the human orphan nuclear receptor, steroid and xenobiotic receptor mediated transcription. Eur J Endocrinol 2001;145 (4):513-517. ##LeCluyse EL. Pregnane X receptor: molecular basis for species differences in CYP3A induction by xenobiotics. Chem Biol Interact 2001;134(3): 283-289. ##Lee JS, Surh YJ. Nrf2 as a novel molecular target for chemoprevention. Cancer Lett 2005;224(2): 171-184. ##Keum YS, Jeong WS, Kong AN. Chemoprevention by isothiocyanates and their underlying molecular signaling mechanisms. Mutat Res 2004;555(1-2): 191-202. ##Milner JA. A historical perspective on garlic and cancer. J Nutr 2001;131(3s):1027S-1031S. ##Bartsch H. DNA adducts in human carcinogenesis: etiological relevance and structure activity relation-ship. Mutat Res 1996;340(2-3):67-79. ##Chao EC, Lipkin SM. Molecular models for the tissue specificity of DNA mismatch repair deficient carcino-genesis. Nucleic Acids Res 2006;34(3):840-852. ##Cooke MS, Evans MD, Dove R, Rozalski R, Gack-owski D, Siomek A, et al. DNA repair is respon-sible for the presence of oxidatively damaged DNA lesions in urine. Mutat Res 2005; 574(1-2):58-66. ##Sancar A, Lindsey-Boltz LA, Unsal-Kacmaz K, Linn S. Molecular mechanisms of mammalian DNA repair and the DNA damage checkpoints. Annu Rev Biochem 2004;73:39-85. ##Frisco S, Choi SW. Gene-nutrient interactions in one carbon metabolism. Curr Drug Metab 2005;6 (1):37-46. ##Astley SB, Elliott RM, Archer DB, Southon S. Evidence that dietary supplementation with caro-tenoids and carotenoid rich food modulate the DNA damage: repair balance in human lympho-cytes. Br J Nutr 2004;91(1):63-72. ##Moller P, Loft S. Interventions with anti oxidants and nutrients in relation to oxidative DNA damage and repair. Mutat Res 2004;551(1-2):79-89. ##Jacks T, Weinberg RA. Taking the study of cancer cell survival to a new dimension. Cell 2002;111 (7):923-925. ##Chen C, Kong AN. Dietary cancer chemo preven-tive compounds: from signaling and gene expres-sion to pharmacological effects. Trends Pharmacol Sci 2005;26(6):318-326. ##Weinstein IB. Disorders in cell circuitry during multistage carcinogenesis: the role of homeostasis. Carcinogenesis 2000;21(5):857-864. ##Agarwal R. Cell signaling and regulators of cell cycle as molecular targets for prostate cancer prevention by dietary agents. Biochem Pharmacol 2000;60(8):1051-1059. ##Poli G, Leonarduzzi G, Biasi F, Chiarpotto E. Oxidative stress and cell signaling. Curr Med Chem 2004;11(9):1163-1182. ##Martin KR. Targeting apoptosis with dietary bio-active agents. Exp Biol Med 2006;231(2):117-129. ##Hu R, Kong AN. Activation of MAP kinases, apoptosis and nutrigenomics of gene expression elicited by dietary cancer-prevention compounds. Nutrition 2004;20(1):83-88. ##Krammer PH. CD95's deadly mission in the immune system. Nature 2000;407(6805):789-795. ##Watson RW, Fitzpatrick JM. Targeting apoptosis in prostate cancer: focus on caspases and inhibitors of apoptosis proteins. BJU Int 2005;96(Suppl 2): 30-34. ##Thornberry NA, Lazebnik Y. Caspases: enemies within. Science 1998;281(5381):1312-1316. ##Hofseth LJ, Ying L. Identifying and defusing weapons of mass inflammation in carcinogenesis. Biochem Biophys Acta 2006;1765(1):74-84. ##Hussain SP, Hofseth LJ, Harris CC. Radical causes of cancer. Nat Rev Cancer 2003;3(4):276-285. ##Storz P. Reactive oxygen species in tumor progres-sion. Front Biosci 2005;10:1881-1896. ##Poli G, Leonarduzzi G, Biasi F, Chiarpotto E. Oxidative stress and cell signaling. Curr Med Chem 2004;11(9):1163-82. ##Shany S, Levy Y, Lahav-Cohen M. The effects of 1alpha, 24 (S)-dihydroxyvitamin D (2) analog on cancer cell proliferation and cytokine expression. Steroids 2001;66(3-5):319-325. ##Chan MM. Inhibition of tumor necrosis factor by curcumin, a phytochemical. Biochem Pharmacol 1995;49(11):1551-1556. ##Xagorari A, Papapetropoulos A, Mauromatis A, Economou M, Fotsis T, Roussos C. Luteolin inhibits an endotoxin-stimulated phosphorylation cascade and proinflammatory cytokine production in macrophages. J Pharmacol Exp Ther 2001;296 (1):181-187. ##Davis JN, Kucuk O, Djuric Z, Sarkar FH. Soy isoflavone supplementation in healthy men prevents NF-kappaB activation by TNF-alpha in blood lymphocytes. Free Radic Biol Med 2001;30 (11):1293-1302. ##Manna SK, Aggarwal BB. All-trans-retinoic acid up regulates TNF receptors and potentiates TNF-induced activation of nuclear factors kappa B, activated protein-1 and apoptosis in human lung cancer cells. Oncogene 2000;19(17):2110-2119. ##Fayette J, Soria JC, Armand JP. Use of angio-genesis inhibitors in tumor treatment. Eur J Cancer 2005;41(8):1109-1116. ##Presta M, Dell’Era P, Mitola S, Moroni E, Ronca R, Rusnati M. Fibroblast growth factor/ fibroblast growth factor receptor system in angiogenesis. Cytokine Growth Factor Rev 2005;16(2):159-178. ##Albini A, Tosetti F, Benelli R, Noonan DM. Tumor inflammatory angiogenesis and its chemo preven-tion. Cancer Res 2005;65(23):10637-10641. ##Pfeffer U, Ferrari N, Morini M, Benelli R, Noonan DM, Albini A. Antiangiogenic activity of chemo preventive drugs. Int J Biol Markers 2003;18(1): 70-74. ##Cockett MI, Murphy G, Birch ML, O’Connell JP, Crabbe T, Millican AT, et al. Matrix metalloprotei-nases and metastatic cancer. Biochem Soc Symp 1998;63:295-313. ##Ii M, Yamamoto H, Adachi Y, Maruyama Y, Shinomura Y. Role of matrix metalloproteinase-7 (matrilysin) in human cancer invasion, apoptosis, growth, and angiogenesis. Exp Biol Med 2006;231 (1):20-27. ##Rose DP, Connolly JM. Regulation of tumor angiogenesis by dietary fatty acids and eico-sanoids. Nutr Cancer 2000;37(2):119-127. ##Cao Y, Cao R, Brakenhielm E. Antiangiogenic mechanisms of diet-derived polyphenols. J Nutr Biochem 2002;13(7):380-390. ##Dulak J. Nutraceuticals as anti angiogenic agents: hope and reality. J Physiol Pharmacol 2005;56 (Suppl 1):51-67. ##Oak MH, El Bedoui J, Schini-Kerth VB. Anti angiogenic properties of natural polyphenols from red wine and green tea. J Nutr Biochem 2005;16 (1):1-8. ##

Generation and Characterization of Mouse Hybridomas Secreting Monoclonal Antibodies Specific for Human IgG3 2 2 Mammalians express several subclasses of the IgG molecule. In human being there are four homologous IgG subclasses, each of which is structurally unique and has different functions. Quantification of IgG subclasses is fundamental to clinical assessment and diagnosis of many diseases as such assessments depends on the availability of subclassspecific antibodies (Abs), particularly monoclonal antibodies (MAbs). In the present study, we produced and characterized two murine MAbs specific for human IgG3 molecule. These MAbs were obtained by the fusion of myeloma cells with splenocytes from Balb/c mice immunized with heavy chain of a human IgG3 myeloma protein. Fused cells were selected in hypoxanthine, aminopterine and thymidine (HAT) medium and cloned by limiting dilution assay. Ab-secreting cells were screened by enzyme-linked immunosorbent assay (ELISA) and the specificity of secreted MAbs was further analyzed, using a panel of purified myeloma proteins by ELISA and immunoblotting. Two stable hybridomas designated 1F18G7 and 1F18A11 were obtained secreting MAbs specific for Fc fragment of human IgG3. None of these MAbs showed cross-reactivity with other immunoglobulin isotypes derived from human and nine other animals, except 1F18A11 which displayed a weak cross-reactivity with only dog serum. Immunoblotting results indicate that these MAbs react with linear epitope(s) located in the heavy chain of human IgG3 molecules. The affinity constant of 1F18G7 and 1F18A11 MAbs was found to be 0.81×109 Mol –1 and 0.71×109 Mol –1, respectively, as measured by ELISA. These two MAbs with relatively high affinity can be useful tools for quantification of IgG3 subclass levels in human serum. 19 26 Fatemeh Hajighasemi Department of Immunology, School of Public Health, Tehran University of Medical Sciences Department of Immunology, School of Medicine, Shahed University Department of Immunology, School of Public Health, Tehran University of Medical Sciences Department of Immunology, School of Medicine, Shahed University IranIran Fazel Shokri Department of Immunology, School of Public Health, Tehran University of Medical Sciences Monoclonal Antibody Research Center, Avecinna Research Center ACECR Department of Immunology, School of Public Health, Tehran University of Medical Sciences Monoclonal Antibody Research Center, Avecinna Research Center ACECR IranIran Heavy chainHuman IgG3ImmunoglobulinIsotypeLight chainMonoclonal antibody 3.pdf Stanisic DI, Richards JS, McCallum FJ, Michon P, King CL, Schoepflin S. IgG subclassspecific responses against Plasmodium falciparum merozoite antigens are associated with control of parasitemia and protection from symptomatic illness. Infect Immun 2009;77(3):1165-1174. ##Murphy SL, Li H, Mingozzi F, Sabatino DE, Hui DJ, Edmonson SA. Diverse IgG subclass responses to adeno-associated virus infection and vector administration. J Med Virol 2009;81(1): 65-74. ##Candore G, Campagna AM, Cuppari I, Di Carlo D, Mineo C, Caruso C. Genetic control of im-mune response in carriers of the 8.1 ancestral haplotype: correlation with levels of IgG subclasses: its relevance in the pathogenesis of autoimmune diseases. Ann N Y Acad Sci 2007; 1110:151-158. ##Towfighi F, Gharagozlou S, Sharifian RA, Kazemnejad A, Esmailzadeh K, Managhchi MR. Comparative measurement of anti factor VIII antibody by Bethesda assay and ELISA reveals restricted isotype profile and epitope specificity. Acta Haematol 2005;114(2):84-90. ##Izad M, Jefferis R, Shokri F. IgG subclass analysis of affinity purified anti-HBs antibodies isolated from normal individuals vaccinated with recombinant HBsAg or naturally infected with hepatitis B virus. Ir J Med Sci 2000;25:42-49. ##Shokrgozar MA, Shokri F. Subtype specificity of anti-HBs antibodies produced by human B-cell lines isolated from normal individuals vaccinated with recombinant hepatitis B vaccine. Vaccine 2002;20(17-18):2215-2220. ##Pasha RP, Bahrami ZS, Niroomanesh S, Ramzi F, Razavi AR, Shokri F. Specificity and isotype of Rh specific antibodies produced by human B-cell lines established from alloimmunized Rh negative women. Transfus Apher Sci 2005;33(2):119-127. ##Wu Q, Zhang Y, Liu M, Wang B, Liu S, He C. Correlation of FcgammaRIIa (CD32) Polymor-phism and IgG antibody subclasses in hemolytic disease of newborn. Neonatology 2009;96(1):1-5. ##Windle HJ, Ang YS, Athie-Morales V, McManus R, Kelleher D. Human peripheral and gastric lymphocyte responses to Helicobacter pylori NapA and AphC differ in infected and uninfected individuals. Gut 2005;54(1):25-32. ##Morais CG, Soares IS, Carvalho LH, Fontes CJ, Krettli AU, Braga EM. IgG isotype to C terminal 19 kD of Plasmodium vivax merozoite surface protein 1 among subjects with different levels of exposure to malaria in Brazil. Parasitol Res 2005;95(6):420-426. ##Burton DR, Gregory L, Jefferis R. Aspects of the molecular structure of IgG subclasses. Monogr Allergy 1986;19:7-35. ##Van de Winkel JG, Anderson CL. Biology of human immunoglobulin G Fc receptors. J Leukoc Biol 1991;49(5):511-524. ##Xu Y, Oomen R, Klein MH. Residue at position 331 in the IgG1 and IgG4 CH2 domains contributes to their differential ability to bind and activate complement. J Biol Chem 1994;269(5): 3469-3474. ##Brekke OH, Michaelsen TE, Sandlie I. The structural requirements for complement activation by IgG: does it hinge on the hinge? Immunol Today 1995;16(2):85-90. ##Lu Y, Harding SE, Michaelsen TE, Longman E, Davis KG, Ortega A. Solution conformation of wild-type and mutant IgG3 and IgG4 immuno-globulins using crystallohydrodynamics: possible implications for complement activation. Biophys J 2007;93(11):3733-3744. ##McLean GR, Torres M, Trotter B, Noseda M, Bryson S, Pai EF. A point mutation in the Ch3 domain of human IgG3 inhibits antibody secre-tion without affecting antigen specificity. Mol Immunol 2005;42(9):1111-1119. ##Edelman GM, Marchalonis JJ. Methods used in studies of the structure of immunoglobulins. In: Williams CH, Chase MW, (eds). Methods of Immunology and Immunochemistry. New York: Academic Press;1967,405-424. ##Loirat MJ, Gourbil A, Frioux Y, Muller JY, Blan-chard D. A murine monoclonal antibody directed against the Gerbich 3 blood group antigen. Vox Sang 1992;62(1):45-48. ##Hajighasemi F, Saboor-Yaraghi AA, Shokri F. Measurement of affinity constant of antihuman IgG monoclonal antibodies by an Elisa based method. Iranian J Immunol 2004;1(3):154-161. ##Beatty JD, Beatty BG, Vlahos WG. Measurement of monoclonal antibody affinity by non-competi-tive enzyme immunoassay. J Immunol Meth 1987;100(1-2):173-179. ##Hadji-Ghasemi F, Gharagozlou S, Ghods R, Roohi A, Khoshnoodi J, Shokri F. Generation and characterization of a mouse monoclonal antibody with specificity similar to staphylococcal protein A (SPA). Hybrid Hybridomics 2003;22(1):33-39. ##Reimer CB, Phillips DJ, Aloisio CH, Moore DD, Galland GG, Wells TW. Evaluation of thirty one mouse monoclonal antibodies to human IgG epitopes. Hybridoma 1984;3(3):263-275. ##Jefferis R, Reimer CB, Skvaril F, de Lange G, Ling NR, Lowe J. Evaluation of monoclonal antibodies having specificity for human IgG subclasses: Results of an IUIS/WHO collabora-tive study. Immunol Lett 1985:10(3-4):223-252. ##Jefferis R, Reimer CB, Skvaril F, de Lange GG, Goodall DM, Bentley Tl, et al. Evaluation of monoclonal antibodies having specificity for human IgG subclasses: results of the 2nd IUIS/WHO collaborative study. Immunol Lett 1992;31(2):143-168. ##Hasnaoui M, Blanchard D, Willem C, Loirat MJ, Lambin P. Production and properties of mono-clonal antibodies against human IgG isotypes. Hybridoma 1996;15(5):351-358. ##Hamilton RG, Morrison SL. Epitope mapping of human immunoglobulin-specific murine mono-clonal antibodies with domain- switched, deleted and point-mutated chimeric antibodies. J Im-munol Meth 1993;158(1):107-122. ##Jefferis R, Reimer C, Skvaril F, De Lange GG. WHO/IUIS program for the standardization of immunological reagents: availability of mouse monoclonal antibodies (MAb) to Human IgG subclasses. Immunology 1988;63:171. ##Recht B, Frangione B, Franklin E, Van LE. Struc-tural studies of a human ?3 myeloma protein (Goe) that binds Staphylococcal protein A. J Immunol 1981;127(3):917-923 . ##Phillips DJ, Wells TW, Reimer CB. Estimation of association constants of 42 monoclonal antibodies to human IgG epitopes using a fluorescent sequential saturation assay. Immunol Lett 1988; 17(2):159-168. ##Mazza G, Whiting AH, Day MJ, Duffus WP. Development of an enzyme-linked immunosor-bent assay for the detection of IgG subclasses in the serum of normal and diseased dogs. Res Vet Sci 1994;57(2):133-139. ##Mazza G, Whiting AH, Day MJ, Duffus WP. Preparation of monoclonal antibodies specific for the subclasses of canine IgG. Res Vet Sci 1994;57 (2):140-145. ##

Conjugation of Monoclonal Antibodies to Super Paramagnetic Iron Oxide Nanoparticles for Detection of her2/neu Antigen on Breast Cancer Cell Lines 2 2 Conjugation of monoclonal antibodies to super paramagnetic nanoparticles is an effective method for cancer diagnosis and treatment. In this study the humanized anti her2/neu monoclonal antibody- Herceptin- was conjugated to super paramagnetic iron oxide (SPIO) nanoparticles using EDC method. The concentration of the conjugated antibodies was measured by Bradford assay. The antibody-nanoparticle conjugates were incubated with SKBR-3 and T47D human breast carcinoma cell lines and the presence of the conjugates on cell surface was confirmed by Prussian blue iron staining method. Conjugation of Herceptin to SPIO resulted in a precipitate-free conjugate containing 20µg antibody/mg SPIO. Prussian blue iron-staining of cells showed successful binding of the conjugates to the cell surfaces. Conjugation of monoclonal antibodies to SPIO may be a useful method for detection of tumor cells, especially by MRI techniques. 27 31 Fereshteh Shamsipour Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Amir-Hassan Zarnani Department of Immunology, Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Department of Immunology, Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Roya Ghods Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Mahmood Chamankhah Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Iran Flora Forouzesh Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Iran Sedigheh Vafaei Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Ali Ahmad Bayat Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Mohammad Mehdi Akhondi Department of Immunology, Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Department of Immunology, Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Mohammad Ali Oghabian Research Centre for Science and Technology in Medicine, Tehran University of Medical Sciences Research Centre for Science and Technology in Medicine, Tehran University of Medical Sciences Iran Mahmood Jeddi-Tehrani Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Breast cancerConjugationHerceptinNanomagnetic particles 4.pdf Santra S, Zhang P, Wang K, Tapec R, Tan W. Conjugation of biomolecules with luminophore-doped silica nanoparticles for photostable biomarkers. Anal Chem 2001;73(20):4988-4993. ##Gruettner C, Teller J, Westphal F, Ivkov R. Magnetic nanoparticle compositions and methods related thereto. Patent application publication 2005. Pub No: US 2005 /0271745 A1 Pub Date: Dec 8, 2005. ##Nagasaki Y, Kobayashi H, Katsuyama Y, Jomura T, Sakura T. Enhanced immunoresponse of antibody mixed-PEG coimmobilized surface construction of high performance immuno-magnetic ELISA system. J Colloid Interface Sci 2007;309(2):524-530. ##Baio G, Fabbi M, d.Totero D, Ferrini S, Cilli M, Derchi LE, Neumaier CE. Magnetic resonance imaging at 1.5T with immunospecific contrast agent in vitro and in vivo in a xeno-transplant model. Mag Reson Mater Phy 2006;19:313-320. ##Itoa A, Kugaa Y, Hondaa H, Kikkawab H, Horiuchib A, Watanabeb Y, et al. Magnetite nanoparticle-loaded anti-her2 immunoliposomes for combination of antibody therapy with hyper-thermia. Cancer Lett 2004;212(2):167-175. ##Kawasaki ES, Player A. Nanotechnology, nano-medicine, and the development of new, effective therapies for cancer. Nanomedicine 2005;1(2): 101-109. ##Lawson EL, Clifton JG, Huang F, Li X, Hixson DC, Josic D. Use of magnetic beads with immo-bilized monoclonal antibodies for isolation of highly pure plasma membranes. Electrophoresis 2006;27(13):2747-2758. ##Lee JH, Huh YM, Jun YW, Seo JW, Jang JT, Song HT, et al. Artificially engineered magnetic nanoparticles for ultra-sensitive molecular imaging. Nat Med 2007;13(1):95-99. ##Gruttnera C, Mullera K, Tellera J, Westphala F, Foremanb A, Ivkovb R. Synthesis and antibody conjugation of magnetic nanoparticles with im-proved specific power absorption rates for alter-nating magnetic field cancer therapy. J Magn Mater 2007;311:181-186. ##Denardo SJ, Denardo G, Miers L, Natarajan A, Foreman A, Gruettner C, et al. Development of tumor targeting bio probes (111In-Chimeric L6 Monoclonal Antibody nanoparticles) for alter-nating magnetic field cancer therapy. Clin Cancer Res 2005;11(19 Pt2):7087s-7092s. ##Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248-254. ##Owen CS, Sykes NL. Magnetic labeling and cell sorting. J Immunol Methods 1984;73(1):41-48. ##Funovicsa MA, Kapellerb B, Hoellerc C, Sud HS, Kunstfeldc R, Puiga S, et al. MR imaging of the her2/neu and 9.2.27 tumor antigens using im-munospecific contrast agents. Magn Reson Im-aging 2004;22(6):843-850. ##Bancroft JD, Stevens A. Theory and Practice of Histological Techniques, 4th ed. New York: Chur-chill Livingstone Inc;1996. ##Luna LG. Manual of Histological Staining Methods of the AFIP. 3rd ed. New York: McGraw Hill; 1968. ##Wagner K, Kautz A, Ro¨ der M, Schwalbe M, Pachmann K, Clement JH, et al. Synthesis of oligonucleotide-functionalized magnetic nano-particles and study on their in vitro cell uptake. Appl Organometal Chem 2004;18(10):514-519. ##Prestvik WS, Berge A, Mork PC, Stenstad PM, Ugelstad J. Preparation and application of mono-sized particles in selective cell separation. In: Hafeli U, Schutt W, Teller J, Zborowski M (eds). Scientific and clinical applications of magnetic carriers. New York: Springer;1997,11-35. ##Hilger I, Trost R, Reichenbach JR, et al. MR imaging of her2/neu protein using magnetic nano-particles. Nanotechnology 2007;18(13):135103-135111. ##Zhang C, Jugold M, Woenne EC, Lammers T, Morgenstern B, et al. Specific targeting of tumor angiogenesis by RGD-conjugated ultra-small superparamagnetic Iron oxide particles using a clinical 1.5-T magnetic resonance scanner. Cancer Res 2007;67(4):1555-1562. ##

Producing a Mammalian GFP Expression Vector Containing Neomycin Resistance Gene 2 2 The green fluorescent protein (GFP) was originally isolated from the Jellyfish Aequorea Victoria that fluoresces green when exposed to blue light. GFP protein is composed of 238 amino acids with the molecular mass of 26.9 kD. The GFP gene is frequently used in cellular and molecular biology as a reporter gene. To date, many bacterial, yeast , fungal, plants, fly and mammalian cells, including human, have been created which express GFP. Martin Chalfie, Osamu Shimomura, and Roger Tsien were awarded the 2008 noble prize in chemistry for their discovery and development of GFP. In many studies on mammalian cells, GFP gene is introduced into cells using vector-based systems or a recombinant virus to track the location of a target protein or to study the expression level of the gene of interest, but in these studies there is no selection marker to normalize transfection. According to the importance of neomycin gene as a selection marker in mammalian cells, we aimed to produce a GFP expression vector that contains neomycin gene. GFP gene was separated from pEGFP-N1 vector and was inserted in the back-bone of pCDNA3.1/His/LacZ vector that contained the neomycin gene. The resulted vector contained GFP beside neomycin gene. 33 36 Manizheh Izadi Department of Medical Genetics, Faculty of Medicine, Tehran University of Medical Science Department of Medical Genetics, Faculty of Medicine, Tehran University of Medical Science Iran Maryam Abiri Department of Medical Genetics, Faculty of Medicine, Tehran University of Medical Science Department of Medical Genetics, Faculty of Medicine, Tehran University of Medical Science Iran Mohammad Keramatipour Department of Medical Genetics, Faculty of Medicine, Tehran University of Medical Science Department of Medical Genetics, Faculty of Medicine, Tehran University of Medical Science Iran ExpressionGreen Flourescent ProteinNeomycinResistance geneVector 5.pdf Prendergast F, Mann K. Chemical and physical properties of aequorin and the green fluorescent protein isolated from Aequorea forskalea. Bio-chemistry 1978;17(17):3448-3453. ##Shimomura O, Johnson F, Saiga Y. Extraction, purification and properties of aequorin, a biolumi-nescent protein from the luminous hydromedusan, Aequorea. J Cell Comp Physiol 1962;59:223-239. ##Prasher D, Eckenrode V, Ward W, Prendergast F, Cormier M. Primary structure of the Aequorea victoria green-fluorescent protein. Gene 1992;111 (2):229-233. ##Ormo M, Cubitt A, Kallio K, Gross L, Tsien R, Remington S. Crystal structure of the Aequorea victoria green fluorescent protein. Science 1996; 273(5280):1392-1395. ##Yang F, Moss L, Phillips G. The molecular structure of green fluorescent protein. Nat Bio-technol 1996;14(10):1246-1251. ##Yuste R. Fluorescence microscopy today. Nat Methods 2005;2(12):902-904. ##Phillips G. Green fluorescent protein-a bright idea for the study of bacterial protein localization. FEMS Microbiol Lett 2001;204(1):9-18. ##Tsien R. The green fluorescent protein. Annu Rev Biochem 1998;67:509-544. ##Chudakov D, Lukyanov S, Lukyanov K. Fluorescent proteins as a toolkit for in vivo imaging. Trends Biotechnol 2005;23(12):605-613. ##

Construction of a High Efficiency PCR Products Cloning T Vector Using pGEM-5zf (+) 2 2 A highly efficient cloning vector was constructed for cloning PCR products by inserting an 80 bp DNA fragment into pGEM-5zf (+) vector. The Xcm I digestion of this vector gave rise to a 3’ overhanging deoxythymidine offering the possibility of cloning PCR products with 3' adenosine overhang created by Taq DNA polymerase. Furthermore, two EcoR I sites were added to the construct for identification of recombinant plasmids using a single restriction enzyme. Taken together, the more efficient cloning performance and the lower cost of this vector as compared to the commercial T vector, suggests that it may be one of the best T vectors for cloning of PCR products. 37 39 Yaofeng Zhao Division of Clinical Immunology, Department of Laboratory Medicine, Karolinska University Hospital Huddinge, SE-141 86 Division of Clinical Immunology, Department of Laboratory Medicine, Karolinska University Hospital Huddinge, SE-141 86 Sweden Zhancai Liu Department of Physics, Chemistry and Biology, Jiaozuo Teachers College, Jiaozuo, 454001 P. R. China Shuyang Yu Division of Clinical Immunology, Department of Laboratory Medicine, Karolinska University Hospital Huddinge, SE-141 86 Division of Clinical Immunology, Department of Laboratory Medicine, Karolinska University Hospital Huddinge, SE-141 86 Sweden Sicheng Wen Division of Clinical Immunology, Department of Laboratory Medicine, Karolinska University Hospital Huddinge, SE-141 86 Division of Clinical Immunology, Department of Laboratory Medicine, Karolinska University Hospital Huddinge, SE-141 86 Sweden Lennart Hammarstrom Division of Clinical Immunology, Department of Laboratory Medicine, Karolinska University Hospital Huddinge, SE-141 86 Division of Clinical Immunology, Department of Laboratory Medicine, Karolinska University Hospital Huddinge, SE-141 86 Sweden Hodjattallah Rabbani Immune and Gene Therapy Laboratory, CCK, Karolinska University Hospital Solna Department of Antigen and Antibody Engineering, Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Immune and Gene Therapy Laboratory, CCK, Karolinska University Hospital Solna Department of Antigen and Antibody Engineering, Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR SwedenIran CloningPCR products pGEM-5zf(+)T vector 6.pdf Clark JM. Novel non-templated nucleotide addition reactions catalyzed by procaryotic and eucaryotic DNA polymerases. Nucleic Acids Res 1988;16(20):9677-9686. ##Holton TA, Graham MW. A simple and efficient method for direct cloning of PCR products using ddT-tailed vectors. Nucleic Acids Res 1991;19(5): 1156. ##Borovkov AY, Rivkin MI. XcmI-containing vector for direct cloning of PCR products. Biotechniques 1997;22(5):812-814. ##Cha J, Bishai W, Chandrasegaran S. New vectors for direct cloning of PCR products. Gene 1993; 136(1-2):369-370. ##Harrison J, Molloy PL,Clark SJ. Direct cloning of polymerase chain reaction products in an XcmI T- vector. Anal Biochem 1994;216(1):235-236. ##Jo C, Jo SA. A simple method to construct T-vec-tors using Xcm I cassettes amplified by nonspe-cific PCR. Plasmid 2001;45(1):37-40. ##Kovalic DJ, Kwak H, Weisblum B. General method for direct cloning of DNA fragments generated by the polymerase chain reaction. Nucleic Acids Res 1991;19(16):4560. ##Mead DA, Pey NK, Herrnstadt C, Marcil RA, Smith LM. A universal method for the direct cloning of PCR amplified nucleic acid. Biotech-nology 1991;9(7):657-663. ##

Assessment of Thyroglobulin Expression in Reproductive Organs at Different Stages of Mouse Estrous Cycle 2 2 Prevalence of abortion is higher in women with autoimmune thyroid disease. In the majority of cases, however, no abnormality of thyroid function is detected despite the high levels of antithyroid antibodies. The direct influence of such harmful autoantibodies in female reproductive organs may serve a role in pregnancy loss. In this study, expression of thyroglobulin in the reproductive tissues of cycling mice has been evaluated. Stages of estrous cycle were determined by cellular morphology and ratio of epithelial cells to leukocytes in vaginal smear of Balb/C mice. At each phase, the mice were sacrificed and their uterus, ovary and fallopian tubes were removed. Expression of thyroglobulin-specific transcript in endometrium was investigated by two sets of primers using reverse transcriptase-polymerase chain reaction (RT-PCR). In addition, expression of thyroglobulin in reproductive tissues was assessed by immunohistochemistry and dot blot analysis. The results showed that thyroglobulin mRNA is not expressed in endometrial tissue of Balb/C mice at any stage of estrous cycle. Immunohistochemical analysis also confirmed that thyroglobulin or its cross reactive-antigens are not expressed at the protein level in the female reproductive organs. The results showed that thyroglobulin was not expressed in the reproductive organs of female mice. It is plausible that antithyroglobulin antibodies could interact with newly-generated antigens during placentation and pregnancy. 41 46 Ali Moravvej Department of Immunology, Faculty of Medicine, Iran University of Medical Sciences Department of Immunology, Faculty of Medicine, Iran University of Medical Sciences Iran Mahmood Jeddi-Tehrani Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Ali Reza Salek Moghaddam Department of Immunology, Faculty of Medicine, Iran University of Medical Sciences Department of Immunology, Faculty of Medicine, Iran University of Medical Sciences Iran Pouneh Dokouhaki Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Mahdi Shekarabi Department of Immunology, Faculty of Medicine, Iran University of Medical Sciences Department of Immunology, Faculty of Medicine, Iran University of Medical Sciences Iran Roya Ghods Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Mahdi Shahbazi Department of Immunology, Faculty of Medicine, Iran University of Medical Sciences Department of Immunology, Faculty of Medicine, Iran University of Medical Sciences Iran Jamileh Ghasemi Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Iran Parivash Danesh Department of Immunology, Faculty of Medicine, Iran University of Medical Sciences Department of Immunology, Faculty of Medicine, Iran University of Medical Sciences Iran Ahmad Reza Mahmoudi Department of Immunology, Faculty of Medicine, Iran University of Medical Sciences Department of Immunology, Faculty of Medicine, Iran University of Medical Sciences Iran Amir-Hassan Zarnani Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Immunology Research Center, Faculty of Medicine, Iran University of Medical Sciences Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR Immunology Research Center, Faculty of Medicine, Iran University of Medical Sciences IranIran Antithyroglobulin antibodyEndometriumEstrous cyclePlacentationRecurrent abortionUterus 7.pdf Regan L, Rai R. Epidemiology and the medical cause of miscarriage. Baillieres Best Pract Res Clin Obstet Gynaecol 2000;14(5):839-854. ##Li TC, Makris M, Tomsu M, Tuckerman E, Laird S. Recurrent miscarriage: etiology, management and prognosis. Hum Reprod Update 2002;8(5): 463-481. ##Matalon ST, Blank M, Levy Y, Carp HJ, Arad A, Burek L, et al. The pathogenic role of antithyro-globulin antibody on pregnancy: evidence from an active immunization model in mice. Hum Reprod 2003;18(5):1094-1099. ##Kutteh WH, Yetman DL, Carr AC, Beck LA, Scott RT. Increased prevalence of antithyroid antibodies identified in women with recurrent pregnancy loss but not in women undergoing assisted reproduc-tion. Fertil Steril 1999;71(5):843-848. ##Tincani A, Rebaioli CB, Frassi M, Taglietti M, Gorla R, Cavazzana I, et al. Pregnancy and autoimmunity: maternal treatment and maternal disease influence on pregnancy outcome. Auto Immun Rev 2005;4(7):423-428. ##Hatasaka HH, Branch DW, Kutteh WH, Scott JR. Autoantibody screening for infertility: explaining the unexplained? J Reprod Immunol 1997;34(2): 137-153. ##Stagnaro-Green A, Glinoer D. Thyroid autoim-munity and the risk of miscarriage. Best Pract Res Clin Endocrinol Metab 2004;18(2):167-181. ##Krassas GE. Thyroid disease and female reproduc-tion. Fertil Steril 2000;74(6):1063-1070. ##Poppe K, Velkeniers B. Female infertility and the thyroid. Best Pract Res Clin Endocrinol Metab 2004;18(2):153-165. ##Poppe K, Glinoer D. Thyroid autoimmunity and hypothyroidism before and during pregnancy. Hum Reprod 2003;9(2):149-161. ##Marai I, Carp H, Shai S, Shabo R, Fishman G, Shoenfeld Y. Autoantibody panel screening in recurrent miscarriages. Am J Reprod Immunol 2004;51(3):235-240. ##Lockshin MD. Autoimmunity, infertility and assisted reproductive technologies. Lupus 2004;13 (9):669-672. ##Pop VJ, de Vries E, van Baar AL, Waelkens JJ, de Rooy HA, Horsten M, et al. Maternal thyroid peroxidase antibodies during pregnancy: a marker of impaired child development? J Clin Endocrinol Metab 1995;80(12):3561-3566. ##Sherer Y, Tartakover-Matalon S, Blank M, Matsura E, Shoenfeldlt Y. Multiple autoantibodies associated with autoimmune reproductive failure. J Assist Reprod Genet 2003;20(2):53-57. ##Shoenfeld Y, Carp HJ, Molina V, Blank M, Cervera R, Balasch J, et al. Autoantibodies and prediction of reproductive failure. Am J Reprod Immunol 2006;56(5-6):337-344. ##King AE, Critchley HO, Kelly RW. Innate immune defenses in the human endometrium. Reprod Biol Endocrinol 2003;1:116. ##Imaizumi M, Pritsker A, Kita M, Ahmad L, Unger P, Davies T. Pregnancy and murine thyroiditis: thyroglobulin immunization leads to fetal loss in specific allogeneic pregnancies. Endocrinology 2001;142(2):823-829. ##

Production and Purification of Streptokinase by Protected Affinity Chromatography 2 2 Streptokinase is an extracellular protein, extracted from certain strains of beta hemolytic streptococcus. It is a non-protease plasminogen activator that activates plasminogen to plasmin, the enzyme that degrades fibrin cloth through its specific lysine binding site; it is used therefore as a drug in thrombolytic therapy. The rate of bacterial growth and streptokinase production was studied in condition of excess glucose addition to culture media and its pH maintenance. The streptokinase product of the bacterial culture was preliminary extracted by salt precipitation and then purified by affinity chromatography on plasminogen substituted sepharose-4B in a condition that the plasminogen active site was protected from streptokinase-induced activation. The purity of streptokinase was confirmed by SDS-PAGE and its biological activity determined in a specific streptokinase assay. The results showed that in the fed –batch culture, the rate of streptokinase production increased over two times as compared with the batch culture while at the same time, shortening the streptokinase purification to a single step increased the yield over 95 % at the chromatography stage. 47 51 Mohammad Babashamsi Department of Medical Diagnostics and Biological Products, Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Department of Medical Diagnostics and Biological Products, Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Mohammad Hossein Razavian Islamic Azad University,Qom branch Islamic Azad University,Qom branch Iran Mohammad Reza Nejadmoghaddam Department of Recombinant Technology, Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Department of Recombinant Technology, Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Iran Affinity chromatographyCulturePlasminogenPurificationStreptokinase 8.pdf Tillet WS, Garner RL. The fibrinolytic activity of hemolytic streptococci’s. J Exp Med 1933;58(4): 485-502. ##Schick LA, Castellino F. Direct evidence for the generation of an active site in the plasminogen moiety of the streptokinase-human plasminogen activator complex. J Biochem Biophys Res Com-mun 1974;57(1):47-54. ##Hoffman R, Benz EJ Jr, Shattil SJ. Hematology: Basic principles and practice (Eds). New York: Churchill Livingstone;1991. ##Rodriguez P, Fuentes P, Barro M, Alvarez JG, Muñoz E, Collen D, et al. Structural domains of streptokinase involved in the interaction with plasminogen. Eur J Biochem 1995;229(1):83-90. ##Hermentin P, Cuesta-Linker T, Weisse J, Schmidt KH, Scheld M, Thimme. M. Comparative analysis of the activity and content of different strepto-kinase preparations. Eur H J 2005;26(9):933-940. ##Banerjee A, Chistic Y, Banerjee UC. Strepto-kinase, a clinically useful thrombolytic agent. Bio-technol Adv 2004;22(4):287-307. ##Erdogan S, Ozer AY, Volkan B, Caner B, Bilgili H. Thrombus localization by using streptokinase containing vesicular systems. Drug Deliv 2006;13 (4):303-309. ##Jackson KW, Tang J. Complete amino acid sequence of streptokinase and its homology with serine proteases. Biochemistry 1982;21(26):6620-6625. ##Karush F, Lacocca VF, Harris TN. Growth of group a hemolytic streptococcus in the steady state. J Bacteriol 1956;72(3):283-294. ##Ogburn CA, Harris TN, Harris S. Extra cellular antigens in steady-state cultures of the hemolytic streptococcus: Production of proteinase at low pH. J Bacteriol 1958;76(2):142-151. ##Rosenberger RF, Elsden SR. The yields of Strepto-coccus faecalis grown in continuous culture. J Gen Microbiol 1960;22:726-739. ##Blatt WF, Segal H, Gray JL. Purification of streptokinase and human plasmin and their inter-action. Thromb Diath Haemorrh 1964;11:393-403. ##De Renzo EC, Siiteri PK, Hutchings BL, Bell PH. Preparation and certain properties of highly purified streptokinase. J Biol Chem 1967;242(3): 533-542. ##Dillon HC Jr, Wannamaker LW. Physical and immunological differences among streptokinases. J Exp Med 1965;121;351-371. ##Castellino FJ, Sodetz JM, Brockay WJ, Siefring GE. Streptokinase. Methods Enzymol 1976;45:244-257. ##Taylor FB, Botts J. Purification and charac-terization of streptokinase with studies of strepto-kinase activation of plasminogen. Biochemistry 1968;7(1):232-242. ##Jeong IK, Young EL, Jeung SJ, Myung B. Purification of streptokinase by affinity chroma-tography using human plasminogen. Korean Biochem J 1993;26(2):172-175. ##Nejadmoghaddam MR, Modarresi MH, Baba shamsi M, Chamankhah M. Cloning and over ex-pression of active recombinant fusion strepto-kinase: a new approach to facilitate purification. Pak J Biol Sci 2007;10(13):2146-2151. ##Babashamsi M. Immobilization of plasminogen in agarose for rapid detection of streptokinase. Iranian Biomed J 2000;4:170. ##Jackson KW, Esmon N, Tang J. Streptokinase and staphylokinase. Methods Enzmol 1981;80:387-394. ##Deutsch DG, Mertz ET. Plasminogen purification from human plasma by affinity chromatography. Science 1970;170(962):1095-1096. ##