Biomedical Technology Reserch Center

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Design and Production:

Printing SectionPrinted and Multimedia Division

National Science and Technology Development Agency

Tel: +66 2564 7000 ext: 1162

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Research StrategyIn the Biomedical Technology Research Center, both basic and applied researches are carried out. The research strategies are that various technologies for production of recombinant protein antigens are established. The established technologies are then applied to produce protein of interests and used as immunogen for both polyclonal antibody and monoclonal antibody production. The genetic engineered antigens are generated using various platforms, i.e. bacterial and mammalian cell system. In addition, recombinant antibodies are also engineered in some cases.

The produced antigens and antibodies are further exploited in basic researches on leukocyte surface molecules, tumor biology, thalassemia, and gene therapy. In addition, the produced antigens and antibodies are employed for development immunodiagnostic kits for several emerging diseases in Thailand, including AIDS, thalassemia, and cancers.

As the research center carries several effective methods for antigen and antibody production, publicity service for antigen and antibody productions are provided.

Biomedical Technology Reserch Center

National Center for Genetic Engineering and Biotechnology

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Advance Molecular biology Technology: : Recombinant proteins (various types)

Antibody Technology: Polyclonal antibodies, Monoclonal antibodies,

Recombinant antibodies

Recombinant proteins/Antibodies:Basic Researches/Applied Researches

Basic Researches

- Technology: Antigen/Antibody production- Leukocyte surface molecules/Immune responses- Cancer- Intrabody technology- Gene therapy/Immunotherapy

Applied Researches

- Immunodiagnosis for Thalassemia- Immunodiagnosis for Cancer- Monitoring of small molecules and drugs- Reagent for CD4+ lymphocyte count- ABO reagent- Reagent for determine drug resistant leukemia

Service:

Antigen and Antibody production

National Center for Genetic Engineering and Biotechnology

The overall research strategies of the Biomedical Technology Research Center are as follows:

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Biomedical Technology Reserch Center

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Col laborative Network

Internally

• Medical Biotechnology Unit, Faculty of Medicine, Siriraj hospital,MahidolUniversity

• Thalassemia Research Center, Institute of Science and Technology Development,MahidolUniversity

• Department of Microbiology, Faculty of Medicine, Chiang Mai University

• Department of Biochemistry, Faculty of Medicine, Chiang Mai University

Internationally

• Department of Molecular Immunology, Center for Biomolecular Medicine and Pharmacology, Medical UniversityofVienna,Austria

• Institute of Molecular Genetics,Czech Republic

• Department of Physiology, Manitoba Institutes of Cell Biology, CancerCare Manitoba,UniversityofManitoba,Canada

• Department of Molecular Biology,The Scripps Research Institute, USA

• CenterforVaccinesandImmunityColumbus Children’s Research Institute,USA

• DepartmentofBiochemistry,University Paris Sud 11, France

• Department of Pathology, University ofWashington,USA

• Laboratoire de Virologie et Pathogénèse Virale, UniversitéClaude Bernard de Lyon, France

Biomedical Technology Reserch Center

National Center for Genetic Engineering and Biotechnology

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National Center for Genetic Engineering and Biotechnology

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Awards• Outstanding Scientist Awards

2008 from the Foundation for the Promotion of Science and Technology under the Patronage of His Majesty the King (Prof. Dr. Watchara Kasinrerk)

• Mahidol B. Braun Award 2007 fromMahidolUniversity (Prof.Dr.Watchara Kasinrerk)

• National Innovation Award 2007 (the Second Level) from The National Innovation Agency for the project entitled “Development of Reagent for Enumeration of CD4+ Lymphocytes” (Prof. Dr. Watchara Kasinrerk)

• ChiangMaiUniversityAlumniAward 2007 from Chiang Mai University (Prof. Dr. Watchara Kasinrerk)

• Chiang Mai University AlumniAward2007fromFacultyofMedical Technology, Mahidol University (Assoc. Prof. Dr. Chatchai Tayapiwatana)

• The Best Thesis Advisor Award2007 from Graduate School,Chiang Mai University for theproject entitled “Development of Enzyme-Linked Immunosorbent Assay and Immunochromatographic Assay for the Detection of Salbutamol” (Assoc. Prof. Dr. Chatchai Tayapiwatana)

• Chiang Mai University Golden Elephant Award 2006 (The best Teacher Award) from Chiang Mai University (Prof. Dr. Watchara Kasinrerk)

• The Best Thesis Supervisor (Master degree) Award 2006 from Graduate School, Chiang Mai University for the project entitled “Production of Monoclonal Antibodies against Hemoglobins of Bart’s Hydrops Fetalis and its Applications” (Prof. Dr. Watchara Kasinrerk)

• OutstandingResearchoftheYear2006 from the Thailand Research Fund for the project entitled “Development of Reagent for Enumeration of CD4+ Lymphocytes” (Prof. Dr. Watchara Kasinrerk)

• AAI Junior Faculty Award 2006 for the Annual Meeting of AAI 2006 in Boston, USA from the American Association of Immunologists for the project entitled “Binding of Multivalent CD147 Phage Induces Apoptosisof U937 Cells” (Assoc. Prof. Dr.Chatchai Tayapiwatana)

• Young Scientist Award 2006 for the 22nd National Congress on Allergy and Immunology from Thai Society of Allergy and Immunology for the project entitled “Applications of Intrabodies for Immunotherapy: a Current Approach” (Assoc. Prof. Dr. Chatchai Tayapiwatana)

Biomedical Technology Reserch Center

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Research Staffs

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Prof. Watchara Kasinrerk, Ph.D.Director(Immunology,UneversitatfurBodenkuturWien,Austria)

Assoc. Prof. Chatchai Tayapiwatana, Ph.D.Deputy Director(MolecularBiology,ChiangMaiUniversity,Thailand)

Assoc. Prof. Tanusak Tatu, Ph.D.(BiochemistryandMolecularGenetics,UniversityofOxford,UK)

Assist. Prof. Rachada Cressey, Ph.D.(MolecularOncology,UniversityofNewcastleUponTyne,UK)

Assist. Prof. Sawitee Chaimpanichayakul, Ph.D.(Biochemistry,ChiangMaiUniversity,Thailand)

Tanawan Samleeral, Ph.D.(BiomedicalSciences,ChiangMaiUniversity,Thailand)

Khanittha Taneyhill, Ph.D.(Biochemistry,ChiangMaiUniversity,Thailand)

Nutjeera Intasai, Ph.D.(MedicalTechnology,MahidolUniversity,Thailand)

Kajornsal Tragoolpua, Ph.D.(BiomedicalSciences,ChiangMaiUniversity,Thailand)

Bordin Butr-indr, Ph.D.(Biotechnology,ChiangMaiUniversity,Thailand)

Biomedical Technology Reserch Center

National Center for Genetic Engineering and Biotechnology

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National Center for Genetic Engineering and Biotechnology

2005

• Boonmuen, S., Tayapiwatana, C. and Kasinrerk, W. 2005. Comparison of polyclonalanti-CD147antibodyproduction using DNA based and phage-displayed CD147immunizations.Sci Asia,31:83-86.

• Cressey, R., Watananupong, O., Lertprasertsuke, N. and Vinitketkumnuen, U. 2005. Alteration of protein expression pattern of vascular endothelial growth factor (VEGF) from soluble to cell- associated isoform during tumourigenesis. BMC Cancer,5:128-135.

• Punturee, K., Wild, CP., Kasinrerk, W. and Vinitketkumnuen, U. 2005. Immunomodulatory activities of Centella asiatica and Rhinacanthus nasutus extracts. Asian Pac J Cancer Prev,6(3): 396-400.

2006

• Anuchapreeda, S., Thanarattanakorn,P., Sittipreechacharn, S., Chanarat, P. and Limtrakul P. 2006. Curcumin inhibits WT1 gene expression in human leukemic K562 cells. Acta Pharmacol Sin, 27(3):360-366.

• Anuchapreeda, S., Limtrakul, P., Thanarattanakorn, P., Sittipreechacharn, S. and Chanarat P. 2006. Inhibitory effect of curcumin on WT1 gene expression in patient leukemic cells. Arch Pharm Res,29(1):80-87.

• Anuchapreeda, S., Thanarattanakorn, P., Sittipreechacharn, S., Tima, S., Chanarat, P. and Limtrakul, P. 2006. Inhibitory effect of curcumin on MDR1 gene expression in patient leukemic cells. Arch Pharm Res, 29(10):866-873.

• Anuchapreeda, S., Thanarattanakorn, P., Sittipreechacharn, S., Tima, S., Chanarat, P. and Limtrakul, P. 2006. Inhibitory effect of curcumin on MDR1 gene expression in patient leukemic cells. Arch Pharm Res,29(10):866-873.

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Publications

• Avirutnan, P., Punyadee, N., Noisakran, S., Komoltri, C., Thiemmeca, S., Auethavornanan, K., Jairungsri, A.,Kanlaya, R., Tangthawornchaikul, N., Puttikhunt, C., Pattanakitsakul, S., Yenchitsmanus, P., Mongkolsapaya, J., Kasinrerk, W., Sittisombut, N., Husmann, M., Blettner, M., Vasanawathana, S., Bhakdi, S. and Malasit, P. 2006. Vascular leakage in severe dengue virus infections: a potential role for the non-structural viral protein NS1 and complement. J Infect Dis,193(8):1078-88.

• Chiampanichayakul, S., Peng-In, P.,Khunkaewla, P., Stockinger, H. and Kasinrerk, W. 2006. CD147 contains different bioactive epitopes involving the regulation of cell adhesion and lymphocyte activation. Immunobiology, 211(3):167-178.

• Chiampanichayakul, S., Khunkaewla, P. and Kasinrerk, W. 2006. Na, K ATPase β3 subunit (CD298): Association with α subunit and expression on peripheral blood cells. Tissue Antigens, 68(6):509-17.

• Cressey, R., Pimpa, S., Tontrong, W., Watananupong, O. and Lertprasertsuke, N. 2006. Overexpression of cyclooxygenase-2isassociatedwithp53accumulation and hdm2 overexpression in colorectal adenocarcinoma. Cancer Lett,233(2):232-239.

• Edwards, M., Wong, SC., Chotpadiwetkul, R., Smirlis, D., Phillips, IR. and Shephard, EA. 2006. Transfection of primary cultures of rat hepatocytes. Methods Mol Biol,320:273-82.

• Intasai, N., Mai, S., Kasinrerk, W. andTayapiwatana, C. 2006. Binding of phage displaying multimeric CD147on U937 cell conducts apoptotic signal. Int J Immunol,18:1159–1169.

• Jintaridtha, P., Srisomsapb, C., Vichittumarosc, K., Kalpravidha, R.,

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Winichagoonc, P., Fucharoen, S., Svastib, J. and Kasinrerk, W. 2006. Chicken egg yolk antibodies specific for the gamma (γ) chain of human hemoglobin for diagnosis of Thalassemia. Int J Hematol,83:408-414.

• Nouanthong, P., Pata, S., Sirisanthana, T. and Kasinrerk, W. 2006. A simple manual rosetting method for absolute CD4+ lymphocyte counting in resource-limited countries. Clin Vaccine Immunol,13(5):598-601.

• Tayapiwatana, C., Chotpadiwetkul,R. and Kasinrerk, W. 2006. A novel approach of using streptavidin magnetic bead sorting in vivo biotinylated survivin for monoclonal antibody production. J Immunol Methods,317:1-11.

• Thammawong, P., Kasinrerk,W., Turner, RC. and Tayapiwatana, C. 2006. Twin-arginine signal peptide attributes effective displaying of CD147 on filamentous phage. Appl Microbiol Biotechnol,69(6):697-703.

2007

• Avirutnan, P., Zhang, L., Punyadee,N., Manuyakorn, A., Puttikhunt, C., Kasinrerk, W., Malasit, P., Atkinson, JP. and Diamond, MS. 2007. Secreted non-structural protein NS1 of denguevirus attaches to the surface of cells via interactions with heparan sulfate and chondroitin sulfate. PLoS Pathog, 3(11):1798-1812.

• Cressey, TR., Nangola, S., Tawon, Y., Pattarawarapan, M., Lallemant, M. and Tayapiwatana, C. 2007. Animmunochromatographic strip test for the rapid detection of nevirapine in plasma samples of HIV-infected patients. Antimicrob Agents Chemother, 51:3361-3363.

• Khunkaewla, P., Chiampanichayakul,S., Yasamut, A., Pata, S. andKasinrerk, W. 2007. Production, characterization and functional analysis of newly

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Biomedical Technology Reserch Center

establishedCD99monoclonal antibodies, MT99/1 andMT99/2.Hybridoma, 26(4): 241-249.

• Limjindaporn, T., Netsawang, J., Noisakran, S., Thiemmaca, S., Wongwiwat, W., Sudsaward, S., Avirutnan, P., Puttikhunt, C., Kasinrerk, W., Sirburi, R., Sittisombut,N.,Yenchitsomanus,P. and Malasit, P. 2007. Sensitization to Fas- mediated apoptosis by dengue virus capsid protein. Biochem Biophys Res Commun,362(2):334-339.

• Noisakran, S., Dechtawewat, T.,Rinkaewkan, P., Puttikhunt, C., Kanjanahaluethai, A., Kasinrerk, W., Sittisombut, N. and Malasit, P. 2007. Acterization of dengue virusNS1 stably expressed in 293T cell lines. J Virol Methods,142(1-2):67-80.

• Pattarawarapan,M.,Nangola,S.,Cressey, T.andTayapiwatana,C.2007.Development of a one-step immunochromatographic strip test for the rapid detection of Nevirapine (NVP), a commonly used antiretroviral drug for the treatment of HIV/AIDS.Talanta,71:462-470.

2008

• Chitov, T., Dispan, R. and Kasinrerk,W. 2007. Incidence and diarrhegenic potential of Bacillus cereus in pasteurized milk and cereal products in Thailand. J Food Saf,28:467-481.

• Cressey, R., Pimpa, S., Chewaskulyong, B.,Tayapiwatana, C. and Kasinrerk, W. 2008. Simplified approaches for developing a tumour antigen specific ELISA to detect circulating autoantibodies in cancer patients. BMC Biotechnol, 8(1):16.

• Puttikhunt, C., Keelapang, P., Khemnu, N., Sittisombut, N., Kasinrerk, W. and Malasit, P. 2008. Novel anti-dengue monoclonal antibody recognizing conformational structure of prM-E heterodimeric complex of dengue virus. J Med Virol,80(1):125-133.

Publications

National Center for Genetic Engineering and Biotechnology

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National Center for Genetic Engineering and Biotechnology

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• Sangiambut, S., Keelapang, P., Aaskov, J.,Puttikhunt,C.,Kasinrerk,W.,Malasit, P. and Sittisombut, N. 2008. Multiple regions in dengue virus capsid protein contribute to nuclear localization during virus infection. J Gen Virol, 89: 1254-1264.

• Srithanaviboonchai, K., Rungruengthanakit, K.,Nouanthong,P.,Pata,S.,Sirisanthana, T. and Kasinrerk, W. 2008. Novel low-cost assay for the monitoring of CD4 counts in HIV-infected individuals. J Acquir Immune Defic Syndr, 47(2): 135-139.

• Tragoolpua, K., Intasai, N., Kasinrerk,W.,Mai, S., Yuan, Y. and Tayapiwatana, C. 2008. Generation of functional scFv intrabody to abate the expression of CD147 surface molecule of 293A cells.BMC Biotechnol, 8(1):5.

2009

• Apiratmateekul, N., Phunpea, P. and Kasinrerk, W. 2009. A modified hybridoma technique for production of monoclonal antibodies having desired isotypes. Cytotechnology,60(1-3):53-54.

• Butr-Indr, B., Kasinrerk, W. and Tayapiwatana, C.2009.Sequentialsimplexoptimization of recombinant biotinylated survivin production by Escherichia coli using mineral supplementation. Biochem Eng J, 46(2):115-120.

• Chitov, T., Dispan, R. and Kasinrerk,W. 2008. Incidence and diarrhedenic potential of Bacillus cereus in pasteurized milk and ceral productions in Thailand. J Food Safety,28(4):467-481.

• Intasai, N., Tragoolpua, K., Pingmuang, P., Khunkaewla, P., Moonsom, S., Kasinrerk, W., Lieber, A. and Tayapiwatana, C. 2009. Potent inhibitionofOKT3-inducedT cell proliferation and suppression of CD147 cell surface expression in HeLacells by scFv-M6-1B9. Immunobiology, doi:10.1016/j.imbio.2008.12.006.

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• Limjindaporn, T., Wongwiwat, W., Noisakran,S.,Srisawat,C.,Netsawang,J., Puttikhunt, C., Kasinrerk, W., Avirutnan, P., Thiemmeca, S., Sriburi, R., Sittisombut, N., Malasit, P. and Yenchitsomanus,P.T. 2009. Interaction of dengue virus envelope protein with endoplasmic reticulum-resident chaperones facilitates dengue virus production. Biochem Bioph Res Co,379(2):196-200.

• Noisakran,S.,Dechtawewat,T.,Avirutnan, P., Kinoshita, T., Siripanyaphinyo, U., Puttikhunt, C., Kasinrerk, W., Malasit, P. andSittisombut,N. 2008. Association of dengue virus NS1 protein with lipidrafts. J Gen Virol,89(10):2492-2500.

• Pata,S.,Tayapiwatana,C.andKasinrerk,W. 2009. Three different immunogenpreparation strategies for production of CD4 monoclonal antibodies. Hybridoma, 28(3):159-65.

• Puttikhunt, C., Ong-ajchaowlerd, P., Prommool,T.Sangiambut,S.,Netsawang, J., Limjindaporn, T., Malasit, P. and Kasinrerk, W. 2009. Production and characterization of anti-dengue capsid antibodies suggesting the N terminus egion covering the first 20 amino acids of dengue virus capsid protein is predominantly immunogenic in mice. Arch Virol, 154(8):1211-1221.

• Sakkhachornphop,S.,Jiranusornkul,S.,Kodchakorn,K.,Nangola,S.,Sirisanthana, T.andTayapiwatana,C.2009.Designedzinc finger protein interacting with theHIV-1 integrase recognition sequenceat 2-LTR-circle junctions. Protein Sci, 18(11):2219-2230.

• Tayapiwatana,C.,Kuntaruk,S.,Tatu,T.,Chiampanichayakul, S., Munkongdee, T., Winichagoon, P., Fuchareon, S. and Kasinrerk,W.2009.Simplemethodforscreening of α-thalassaemia 1 carriers. Int J Hematol,89(5):559-567.

Publications