Year 2015, Volume 21 , Issue 7, Pages 325 - 336 2016-01-02

Transfection of Spodoptera frugiperda Insect Cell and Production of Recombinant Protein
Spodoptera frugiperda Böcek Hücrelerinin Transfeksiyonu ve Rekombinant Protein Üretimi

Özlem Üstün AYTEKİN [1]


Rekombinant protein üretimi son yıllarda sıklıkla çalışılan bir konudur. Rekombinant protein üretimi için çalışılan konakçılar arasında kısa ikilenme süresi, protein katlanmasındaki avantajları, süspanse kültürde çalışılabilmesi nedeni ile Spodoptera frugiperda böcek hücre hatları; yüksek verimliliği, büyük gen bölgelerini eksprese etme kapasitesi ve hızlı cevap vermesi nedeni ile de transfeksiyon vektörü olarak baculovirus (BV) tercih edilmektedir. Bu derlemede rekombinant protein üretimini etkileyen bu hücre hatları, üretim ortamları, uygun üretim koşulları, baculovirusler, transfeksiyon metotları, rekombinant protein üretiminde kullanılan biyoreaktör sistemleri, çalışma modları ve biyoreaktör ekipmanları açıklanmaktadır.
Rekombinant protein üretimi son yıllarda sıklıkla çalışılan bir konudur. Rekombinant protein üretimi için çalışılan konakçılar arasında kısa ikilenme süresi, protein katlanmasındaki avantajları, süspanse kültürde çalışılabilmesi nedeni ile Spodoptera frugiperda böcek hücre hatları; yüksek verimliliği, büyük gen bölgelerini eksprese etme kapasitesi ve hızlı cevap vermesi nedeni ile de transfeksiyon vektörü olarak baculovirus (BV) tercih edilmektedir. Bu derlemede rekombinant protein üretimini etkileyen bu hücre hatları, üretim ortamları, uygun üretim koşulları, baculovirusler, transfeksiyon metotları, rekombinant protein üretiminde kullanılan biyoreaktör sistemleri, çalışma modları ve biyoreaktör ekipmanları açıklanmaktadır.
Spodoptera frugiperda, Rekombinant protein üretimi, Baculovirus, Biyoreaktörler
  • Cino J. “High Yield Protein Production from Pichia pastoris Yeast: A Protocol or Benchtop Fermentation”. American Biotechnology Laboratory, 17, 10-13, 1999.
  • Bağış H. “Transgenik Biyoreaktörlerde Rekombinant Proteinlerin Üretimi”. İstanbul Üniversitesi Veteriner Fakültesi Dergisi, 28(1), 113-123, 2002.
  • Üstün-Aytekin Ö, Gürhan İD, Ohura K, Imai T, Öngen G. “Monitoring of the Effects of Transfection with Baculovirus on Sf9 Cell Line and Expression of Human Dipeptidyl Peptidase IV”. Cytotechnology, 66(1), 159-168, 2014.
  • TÜBİTAK. “Vizyon 2023 Projesi Biyoteknoloji ve Gen Stratejileri Grubu, Biyoteknoloji ve Gen Teknolojileri Stratejisi”.http://vizyon2023.tubitak.gov.tr (10.03.2014).
  • Goldschmidt R. “Some Experiments on Spermatogenesis in Vitro”. Proceedings of the National Academy of Sciences of the United States of America, 1(4), 220-222, 1915.
  • Grace TDC. “Establishment of Four Strains of Cells from Insect Tissue Grown in Vitro”. Nature, 195, 788-789, 1962.
  • Lynn DE. “Development of Insect Cell Lines: Virus Susceptibility and Applicability to Prawn Cell Culture”. Methods in Cell Science, 21, 173-81, 1999.
  • Granados RR, McKenna KA. “Insect Cell Culture Methods and Their Use in Virus Research”. Baculovirus Expression Systems and Biopesticides, Wiley-Liss, New York, USA, 1995.
  • Mitsuhashi J. Invertebrate Cell System Applications, FL: CRC Press, Boca Raton, Florida, USA, 1989.
  • Flanders KL. “ACES Publications: Management of Fall Armyworm in Pastures and Hayfields: ANR-1019”. http://www.aces.edu/pubs/docs/A/ANR-1019/index2. tmpl (12.05.2007).
  • Wickham TJ, Davis T, Granados RR, Shuler ML, Wood H. A. “Screening of Insect Cell Lines for the Production of Recombinant Proteins and Infectious Virus in the Baculovirus Expression System”. Biotechnology Progress, 8(5), 391-396, 1992.
  • Hink WF, Bezanson DR, Kurstak E. (Ed.). Invertebrate Cell Culture Media and Cell Lines, Techniques in the Life Science, Elsevier County Clare, Ireland, 1985.
  • Manual, Instruction, Guide to Baculovirus Expression Vector Systems (bevs) and Insect Cell Culture Techniques, Invitrogen Life Technologies, 2011.
  • Mitsuhashi J. “Media for Insect Cell Cultures”. Advances in Cell Culture, 2nd ed. New York, USA, Academic Pres, 1982.
  • Eibl R, Eibl D, Pörtner R, Catapano G, Czermak P. Cell and Tissue Reaction Engineering. 1st ed. New Delhi, India, Springer-Verlag, Berlin Heidelberg, 2009.
  • Richardson CD. Baculovirus Expression Protocols. 1st ed. Totowa, NJ. Humana Press, 1995.
  • Elias CB, Jardin B, Kamen A. “Recombinant Protein Production in Large-Scale Agitated Bioreactors Using the Baculovirus Expression Vector System”. Methods in Molecular Biology, 388, 225-246, 2007.
  • Shao-Hua C, Hong-Liang S, Zuo-Hu L. “Effect of Temperature Oscillation on Insect Cell Growth and Baculovirus Replication”. Applied and Environmental Microbiology, 64(6), 2237-2239, 1998.
  • Adler S, Roy A, Relman S. “Intracellular Acid-Base Regulation. The Interaction Between 𝐶𝑂2 Tension and Extracellular Bicarbonate in the Determination of Muscle Cell pH”. The Journal of Clinical Investigation, 44(1), 21-30, 1965.
  • Taticek R, Petersen S, Konstantinov K, Naveh D. “In Effect of Dissolved Carbon Dioxide and Bicarbonate on Mammalian Cell Metabolism and Recombinant Protein Productivity in High Density Perfusion Culture”. Presented at Cell Culture Engineering VI, San Diego, USA, 1998. [21] deZengotita VM, Schmeizer AE, Miller WM. “Characterization of Hybridoma Cell Response to Elevated pco2 and Osmolality: Intracellular pH, Cell Size, Apoptosis and Metabolism”. Biotechnology and Bioengineering, 77(4), 369-380, 2002.
  • Goss SPA, Singh RJ, Kalyanaraman B. “Bicarbonate Enhances the Peroxidase Activity of Cu, Zn-Superoxide Dismutase”. The Journal of Biological Chemistry, 274(40), 28233-28239, 1999.
  • Vesela A, Wilhelm J. “The Role of Carbon Dioxide in Free Radical Reaction of the Organism”. Physiological Research, 51, 335-339, 2002.
  • Vajrala SG. Mechanism of CO2 Inhibition in Insect Cell Culture, University of Iowa Iowa Research Online. Master Thesis. Lowa University, IA, USA, 2010.
  • Agathos SN. “Insect Cell Bioreactors”. Cytotechnology, 20, 173-189, 1996.
  • Palomares LA, López S, Ramirez OT. “Utilization of Oxygen Uptake Rate to Assess the Role of Glucose and Glutamine in the Metabolism of Infected Insect Cell Cultures”. Biochemical Engineering Journal, 19(1), 87-93, 2004.
  • Pamboukian MM, Jorge SAC, Santos MG, Yokomizo AY, Pereira CA, Tonso A. “Insect Cells Respiratory Activity in Bioreactor”. Cytotechnology, 57(1), 37-44, 2008.
  • Jöbses I, Martens D, Tramper J. “Lethal Events during Gas Sparging in Animal Cell Culture”. Biotechnology and Bioengineering, 37(5), 484-490, 1991.
  • Doran MP. Bioprocess Engineering Principles. 1st ed. UK. Academic Press, 1995.
  • Jain E, Kumar A. “Upstream Processes in Antibody Production: Evaluation of Critical Parameters”. Biotechnology Advances, 26(1), 46-72, 2008.
  • Mirro R, Voll K. “Which Impeller is Right for Your Cell Line? A guide to Impeller Selection for Stirred-Tank Bioreactors”. Bioprocess International, 7, 51-57, 2009.
  • Tramper J, Vlak JM, Gooijer CD. “Scale up Aspects of Sparged Insect-Cell Bioreactors”. Cytotechnology, 20, 221-229, 1996.
  • Kost TA, Condreay JP. “Recombinant Baculoviruses as Mammalian Cell Gene-Delivery Vectors”. Trends in Biotechnology, 20, 173-180, 2002.
  • Oers VMM. “Vaccines for Viral and Parasitic Diseases Produced with Baculovirus Vectors”. Advance in Virus Research, 68, 193-253, 2006.
  • Yin J, Li G, Ren X, Herrler G. “Select What you Need: A Comparative Evaluation of the Advantages and Limitations of Frequently Used Expression System for Foreign Genes”. Journal of Biotechnology, 127(3), 335-347, 2007. Pamukkale Univ Muh Bilim Derg, 21(7), 325-336, 2015 Ö. Üstün Aytekin 335
  • Adams JR, McClintock JT, Adams JR. (Ed.), Bonami JR. (Ed.). Baculoviridae, Nuclear Polyhedrosis Viruses of Insects, Atlas of Invertebrate Viruses, CRC Press, Boca Raton, Florida, 1991.
  • Federici BA. Baculovirus Pathogenesis. 1st ed. New York, USA, Springer Plenum Press, 1997.
  • Mikhailov VS. “Replication of the Baculoviruses Genome”. Molecular Biology, 37(2), 250-259, 2003.
  • Demir İ, Demirbağ ZA. “Protuctive Replication of Hyphantria Cunea Nucleopolyhedrovirus in Lymantria Dispar Cell Line”. Journal of Microbiology and Biotechnology, 16(10), 1485-1490, 2006.
  • Bilimoria SL, Kurstak E. Viruses of Invertebrates, 1st ed. New York, USA, CRC Press, 1991. [41] Kelly DC. “Baculovirus Replication”. Journal of General Virology, 63, 1-13, 1982.
  • Demir İ, Nalçacıoğlu R, Demirbağ Z. “Böcek Virüslerinin Biyoteknolojik Önemi”. Tarım Bilimleri Dergisi, 14(2), 193-201, 2008.
  • Rohrmann GF. Baculovirus Molecular Biology, 3rd ed. Department of Microbiology, Corvallis, USA, Oregon State University, 2011.
  • Madigan MT. Martingo JM. “Brock Biology of Microorganisms”. International Microbiology, 11, 65-73, 2008.
  • Adams NC, Gale NW, Shirley P. (Ed.), Carlos L. (Ed.), Principles and Practice: Mammalian and Avian Transgenesis-New Approaches. Berlin, Germany, Springer-Verlag, 2006.
  • Gülçe İz S, Deliloğlu Gürhan İ. “Temel Hücre Kültürü Kursu”. Ege Üniversitesi Biyomühendislik Bölümü, İzmir, Türkiye, 2010.
  • Çolak A. “Gen Tedavisi”. Türk Nöroşirurji Dergisi, 16(1), 12, 2006.
  • Freshney RI. Culture of Animal Cells. A Manual of Basic Technique. New York, USA, Wiley-Liss, Inc., 2000.
  • Menuel S, Fontanay S, Clarot I, Duval RE, Diez L, Marsura A. “Synthesis and Complexation Ability of a Novel Bis- (guanidinium)-tetrakis-(β-cyclodextrin) dendrimeric Tetrapod as a Potential Gene Delivery (dna and sirna) System, Study of Cellular Sirna Transfection”. Bioconjugate Chemistry, 19(12), 2357-2362, 2008.
  • Freshney RI. Culture of Animal Cells Types: Culture of Animal Cells. 1st ed. New York, USA, John Wiley & Sons, 2005.
  • Layden MJ, Röttinger E, Wolenski FS, Gilmore TD, Martindale MQ. “Microinjection of mRNA or Morpholinos for Reverse Genetic Analysis in the Starlet Sea Anemone Nematostella Vectensis”. Nature Protocols, 8(5), 924-934, 2013.
  • Potter H, Heller R. “Transfection by Electroporation”. Current Protocols in Neuroscience, 57. A1E1-A1E11, 2001.
  • Yamaji H, Tagai S, Fukuda H. “Optimal Production of Recombinant Protein by the Baculovirus-Insect Cell System in Shake-Flask Culture with Medium Replacement”. Journal of Bioscience and Bioengineering, 87(5), 636-641, 1999.
  • Zhang YH, Enden G, Jos´e C. “Insect Cells-Baculovirus System: Factors Affecting Growth and Low MOI Infection”. Biochemical Engineering Journal, 27(1), 8-16, 2005.
  • Carinhas N, Bernal V, Monteiro F, Carrondo MJT, Oliveira R, Alves PM. “Improving Baculovirus Production at High Cell Density Through Manipulation of Energy Metabolism”. Metabolic Engineering, 12(1), 39-52, 2010.
  • Deutschmann SM, Jeager V. “Optimization of the Growth Conditions of Sf21 Insect Cells for High-Density Perfusion Culture in Stirred-Tank Bioreactors”. Enzyme and Microbial Technology, 16(6), 506-512, 1994.
  • Spier RE, Griffiths JB, Berthold W. Animal Cell Technology. 1st ed. San Diego, CA, Academic Press, 1994.
  • Zhang J, Kalogerakis N, Behie LA, Iatrou K. “A Two-Stage Bioreactor System for the Production of Recombinant Proteins Using a Genetically Engineered Baculovirus/Insect Cell System”. Biotechnology Bioengineering, 42(3), 357-366, 1993.
  • Zhang J, Kalogerakis N, Behie LA, Iatrou K. “Optimum Infection Conditions for Recombinant Protein Production in Insect Cell (Bm5) Suspension Culture”. Biotechnology Progress, 10(6), 636-643, 1994.
  • Gooijer CD, Kokenf RHM, Van Lierm FLJ, Kool M, Vlak JM, Tramper J. “A Structured Dynamic Model for the Baculovirus Infection Process in Insect-Cell Reactor Configurations”. Biotechnology and Bioengineering, 40(4), 537-548, 1992.
  • Betenbaugh MJ, Ailor E, Whiteley EM, Hopkins J. Insect Cells and Larvae, Gene Expression Systems. Encyclopedia of Bioprocess Technology, 1st ed. Pennsylvania, USA, Merck and Co., 2002.
  • Van-Lier FLJ, Van-Duijnhoven GC. F., De-Vaan, MMJACM, Vlak JM, Tramper J. “Continuous Beta-Galactosidase Production in Insect Cells with a p10 Gene Based Baculovirus Vector in a Two-Stage Bioreactor System”. Biotechnology Progress, 10(1), 60-64, 1994.
  • Van-Lier FLJ, Van den Hombergh JP, de Gooijer CD, den Boer MM, Vlak JM, Tramper J. “Long-Term SemiContinuous Production of Recombinant Baculovirus Protein in a Repeated (fed-) Batch two-Stage Reactor System”. Enzyme and Microbial Technology, 18(6), 460466, 1996.
  • Hu YC, Wang MY, Bentley WE. “A Tubular Bioreactor for the Infection of Insect Cells with Recombinant Baculovirus”. Cytotechnology, 24(2), 143-152, 1997.
  • Rice JW, Rankl NB, Guranuss TM, Marr CM, Barna JB. Walters MM, Burns DJ. “A Comparison of Large-Scale Sf9 Insect Cell Growth and Protein Production: Stirred Vessel vs. Airlift”. BioTechniques, 15(6), 1052-1059, 1993.
  • King GA, Kuzio J, Daugulis AJ, Faulkner P, Bayly D, Goosen MFA. “Growth of Baculovirus-Infected Insect Cells in Microcapsules to a High Cell and Virus Density”. Biotechnology Letter, 10(10), 683-688, 1988.
  • Taticek RA, Hammer DA, Shuler ML. Baculovirus Expression Systems and Biopesticides. Wiley-Liss, Inc., New York, USA, 1995.
  • Pijlman PG, Vermeesch MGA, Vlak MJ. “Cell Line-Specific Accumulation of the Baculovirus Non-Hr Origin of DNA Replication in Infected Insect Cells”. Journal of Invertebrate Pathology, 83(4), 214-219, 2004.
  • Pijlman PG, Roode CD, Fan X, Roberts OL, Belsham JG, Vlak MJ, Oers van MM. “Stabilized Baculovirus Vector Expressing a Heterologous Gene and gp64 From a Single Bicistronic Transcript”. Journal of Biotechnology, 123(1), 3-21, 2006. Pamukkale Univ Muh Bilim Derg, 21(7), 325-336, 2015 Ö. Üstün Aytekin 336
  • Svensson I. “Physiological Effects of Conditioned Medium and Passage Number on Spodoptera frugiperda Sf9 Serum Free Cultures” Royal Institute of Technology School of Biotechnology Deparment, Stockholm, Sweden, 2005.
  • ContrerasGómez A, Sánchez Mirón A, García Camacho F, Molina Grima E, Chisti Y. “Protein Production Using the Baculovirus Insect Cell Expression System”. Biotechnology Progress, 30(1), 1-18, 2013.
  • Drugmand JC, Schneider YJ. Agathos SN. “Insect Cells as Factories for Biomanufacturing”. Biotechnology Advances, 30(5), 1140-1157, 2012.
  • Bhatia R, Jesionowski G, Ferrance J, Ataai MM. “Insect Cell Physiology”. Cytotechnology, (20), 33-41, 1996.
  • Griffiths B. Animal Cell Products. In: Spier Re (Ed) Encyclopedia of Cell Technology, New York, USA, Wiley, 2000.
  • Chalmers JJ, Shuler ML, Wood HA, Granados RR, Hammer, DA. Baculovirus Expression Systems and Biopesticides. New York, USA, Wiley-Liss, 1995.
  • Tramper J, Williams JB, Joustra D. “Shear Sensitivity of Insect Cells in Suspension”. Enzyme and Microbial Technology, 8(1), 33-36, 1986.
  • Bavarian F, Fan LS, Chalmers JJ. “Microscopic Visualization of Insect Cell-Bubble Interactions. Rising Bubbles, Air-Medium Interface, and the Foam Layer”. Biotechnology Progress, 7(2), 140-150, 1991.
  • Czermak P, Weber C, Nehring D. “A Ceramic Microsparging Aeration System for Cell Culture Reactors”. Publication Series of IBPT-University of Applied Sciences. (1), 1-6, 2005.
  • Doyle A, Griffith J. Cell and Tissue Culture: Laboratory Procedures in Biotechnology. New York, USA, John Wiley and Sons, 1998.
  • Henzler H, Kauling O. “Oxygenation of Cell Cultures”. Bioprocess Engineering, 9(2-3), 61-75, 1993.
  • Varley J, Birch J. “Reactor Design for Large Scale Suspension Animal Cell Culture”. Cytotechnology, 29(3), 177-205, 1999.
  • Chisti Y. “Animal-Cell Damage in Sparged Bioreactors”. Trends in Biotechnology, 18(10), 420-432, 2000.
  • Handa A, Emery AN, Spier RE, “On the Evaluation of Gas-Liquid Interfacial Effects on Hybridoma Viability in Bubble Column Bioreactors”. Developments in Biological Standardization, 66, 241-253, 1987.
  • Handa-Corrigan A, Emery AN, Spier RE. “Effect of Gas-Liquid Interfaces on the Growth of Suspended Mammalian Cells: Mechanisms of Cell Damage by Bubbles”. Enzyme and Microbial Technology, 11(4), 230-235, 1989.
  • Murhammer DW. “The use of Insect Cell Cultures for Recombinant Protein Synthesis: Engineering Aspects”. Applied Biochemistry and Biotechnology, 31, 283-310, 1991.
  • Kretzmer G. “Industrial Processes with Animal Cells”. Applied Microbiology and Biotechnology, 59(2-3), 135-142, 2002.
  • Catapano G, Czermak P, Eibl R, Eibl D, Pörtner R. Bioreactor Design and Scale-Up, Cell and Tissue Reaction Engineering Principles and Practice, Springer-Verlag, Heidelberg, Berlin, 2009.
  • Jain D, Ramasubramanyan K, Gould S, Lenny A, Candelore M, Tola M, Strader C, Alves K, Cuca G, Tung JS, Hunt G, Junker B, Buckland BC, Silberklang M. Large-Scale Recombinant Protein Production using the Insect Cell-Baculovirus Expression Vector System: Antistasin and Beta-Adrenergic Receptor. Editors: Spier RE, Griffiths JB and Meignier B. Production of bioLogicals from Animal Cells in Culture, 345-351, Oxford, ButterworthHeinemann, 1991.
  • Caron AW, Archambault J, Massie B. “High-Level Recombinant Protein Production in Bioreactors Using the Baculovirus Insect Cell Expression System”. Biotechnology and Bioengineering, 36(11), 1133-1140, 1990.
  • Blanchard JH, Ferguson CHR. The Effect of Different Levels of Dissolved Oxygen on Recombinant Protein Production in Sf-9 cells. Editors: Vlak JM, Schlanger E-J, Bernard AR. Baculovirus and Recombinant Protein Production Processes, 247-254, Basel: Editiones Roche, 1992.
  • Schmid G. “Insect Cell Cultivation: Growth and Kinetics”. Cytotechnology, 20, 43-56, 1996.
  • Maranga L, Cunha A, Clemente J, Cruz P, Carrondo MJT. “Scale-Up of Virus-Like Particles Production: Effects of Sparging, Agitation and Bioreactor Scale on Cell Growth, Infection Kinetics and Productivity”. Journal of Biotechnology, 107(1), 55-64, 2004.
  • Oh SKW, Nienow AW, Al-Rubeai M, Emery AN. “The Effects of Agitation Intensity with and Without Continuous Sparging on the Growth and Antibody Production of Hybridoma Cells”. Journal of Biotechnology, 12(1), 45-62, 1989.
  • Kunas KT, Papoutsakis ET. “Damage Mechanisms of Suspended Animal Cells in Agitated Bioreactors with and Without Bubble Entrainment”. Biotechnology and Bioengineering, 36(5), 476-483, 1990
Primary Language en
Journal Section Review Article
Authors

Author: Özlem Üstün AYTEKİN

Dates

Publication Date : January 2, 2016

Bibtex @ { pajes219139, journal = {Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi}, issn = {1300-7009}, eissn = {2147-5881}, address = {}, publisher = {Pamukkale University}, year = {2016}, volume = {21}, pages = {325 - 336}, doi = {}, title = {Transfection of Spodoptera frugiperda Insect Cell and Production of Recombinant Protein}, key = {cite}, author = {AYTEKİN, Özlem Üstün} }
APA AYTEKİN, Ö . (2016). Transfection of Spodoptera frugiperda Insect Cell and Production of Recombinant Protein. Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi , 21 (7) , 325-336 . Retrieved from https://dergipark.org.tr/en/pub/pajes/issue/20562/219139
MLA AYTEKİN, Ö . "Transfection of Spodoptera frugiperda Insect Cell and Production of Recombinant Protein". Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi 21 (2016 ): 325-336 <https://dergipark.org.tr/en/pub/pajes/issue/20562/219139>
Chicago AYTEKİN, Ö . "Transfection of Spodoptera frugiperda Insect Cell and Production of Recombinant Protein". Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi 21 (2016 ): 325-336
RIS TY - JOUR T1 - Transfection of Spodoptera frugiperda Insect Cell and Production of Recombinant Protein AU - Özlem Üstün AYTEKİN Y1 - 2016 PY - 2016 N1 - DO - T2 - Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi JF - Journal JO - JOR SP - 325 EP - 336 VL - 21 IS - 7 SN - 1300-7009-2147-5881 M3 - UR - Y2 - 2020 ER -
EndNote %0 Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi Transfection of Spodoptera frugiperda Insect Cell and Production of Recombinant Protein %A Özlem Üstün AYTEKİN %T Transfection of Spodoptera frugiperda Insect Cell and Production of Recombinant Protein %D 2016 %J Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi %P 1300-7009-2147-5881 %V 21 %N 7 %R %U
ISNAD AYTEKİN, Özlem Üstün . "Transfection of Spodoptera frugiperda Insect Cell and Production of Recombinant Protein". Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi 21 / 7 (January 2016): 325-336 .
AMA AYTEKİN Ö . Transfection of Spodoptera frugiperda Insect Cell and Production of Recombinant Protein. Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi. 2016; 21(7): 325-336.
Vancouver AYTEKİN Ö . Transfection of Spodoptera frugiperda Insect Cell and Production of Recombinant Protein. Pamukkale Üniversitesi Mühendislik Bilimleri Dergisi. 2016; 21(7): 336-325.