Pharmaceutical Biotechnology
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1 Pharmaceutical Biotechnology Debbie S. Retnoningrum; Catur Riani; Tri Suciati; Heni Rachmawati; Ana Indrayati School of Pharmacy, ITB Introduction 1
2 REFERENCES Glick, BR and JJ Pasternak, 2003, Molecular Biotechnology: Principles and Applications of Recombinant DNA, ASM Press, Washington DC Groves MJ, 2006, Pharmaceutical Biotechnology, 2nd ed., CRC, Taylor & Francis. Brown TA, 2006, Gene Cloning & DNA analysis, Blackwell Publ. Oxford Sven Frokjaer and Lars Hovgaard, 2000, Pharmaceutical Formulation Development of Peptides and Proteins, CRC Press Kayser O. And Muller RH, 2004, Pharmaceutical Biotechnology, Wiley VCH. Grietje Moleme, Dirk K.F.Meijer, Drug Targeting:Organ-spesific strategies, Wiley-VCH, 2001 (HR) Saltzman W. M, Tissue engineering: engineering principles for the design of replacement organs dan tissues, Oxford university press, 2004 (TS) W.W. Minuth, R. Strehl, K. Schumacher, Tissue Engineering: Essentials for daily laboratory work, Wiley-VCH, 2005 (TS) Introduction 2
3 TOPICS 1. Introduction (DSR) 2. Nucleic Acid and Protein, Flow of Genetic Information (DSR) 3. Replication and Polymerase Chain Reaction (PCR) (DSR) Quis 1: topics 2 and 3 4. Recombinant DNA technology (DSR) 5. Manipulation of Gene Expression (DSR) 6. Therapeutic Protein, Vaccine and Technology for Recombinant Protein Production (DSR) Quis 2: topics 4, 5, 6 7. Monoclonal Antibody and Technology for Monoclonal Antibody Production (CR) 8. Mid semester exam Introduction 3
4 TOPICS 9. Protein Isolatian and Purification (AI) 10. Analysis of Recombinant Product (CR) 11. Gene Therapy and Transgenic Plant (CR) 12. Biosimilar Products (CR) 13. Oral Presentation (CR/AI) 14. Formulation of Recombinant Proteins (HR) 15. Targeted Drug Delivery (HR) 16. Tissue Engineering (TS) 17. Final Exam Introduction 4
5 1. Introduction (DSR) General view of pharmaceutical biotechnology, explanation about pharmaceutical biotechnology and its applications in pharmacy field, examples of products produced by recombinant DNA technology Glick, BR and JJ Pasternak, 2003, Molecular Biotechnology: Principles and Applications of Recombinant DNA, ASM Press, Washington DC (hal 3 13) Groves MJ, 2006, Pharmaceutical Biotechnology, 2nd ed., CRC, Taylor & Francis (1 4) Introduction 5
6 2. Nucleic acid and protein, flow of genetic information (DSR) Nucleic acid, polypeptide and protein, transcription and translation in prokaryotes and eukaryotes, role of transcription and translation in molecular biotechnology Post test 1 Introduction 6
7 3. Replication and Polymerase Chain Reaction (PCR) (DSR) Replication in prokaryotes and eukaryotes; basic principle of Polymerase Chain Reaction (PCR), modified PCR Quiz 1: topics 2 and 3 Introduction 7
8 4. Recombinant DNA technology (DSR) Basic principle of recombinant DNA technology, vectors: cloning and expression vectors, strategy for DNA isolation, ligation, transformation, clone characterization Post test 2 Introduction 8
9 Recombinant DNA technology 14 October 1980, price of shares of biotechnology company, Genentech increased considerably from $35 to $89. Genentech: a company specializing in recombinant DNA technology, genetic engineering, gene transplantation and gene cloning. Cloning of insulin gene in Escherichia coli to produce human insulin. Genentech, building 32 Introduction 9
10 Recombinant DNA technology 1973: Stanley Cohen and Herbert Boyer discovered a method to transfer genetic information (gene) from one organism to others. 1976: Herbert Boyer cofound Genentech, the first company in USA which applies Recombinant DNA technology 1978: Somatostatin, a regulator for human growth hormone is the first human protein produced by Recombinant DNA technology Introduction 10
11 Emergence of molecular biotechnology A study of the industrial production of good and services by processes using biological organisms, systems, and processes. Scientific disiplines: microbiology, biochemistry and chemical engineering Raw material UPSTREAM PROCESSING Fermentation and biotransformation Downstream processing Pure product Introduction 11
12 Emergence of molecular biotechnology The use of recombinant DNA technology in biotechnology: Molecular biotechnology Using Molecular biotechnology Creation of new strains / mutants rather than isolation/ screening of strains from nature Cells, plants, animals are used as biological factories Introduction 12
13 Commercialization of molecular biotechnology In 1985: > 400 biotechnology companies names with word gene : Biogen, Amgen, Calgene, Engenics, Genex, and Cangene. Now: > 1300 companies in USA and > 2500 world-wide Introduction 13
14 Scientific disiplines contributes to molecular biotechnology MOLECULAR BIOLOGY MICROBIOLOGY CROPS BIOCHEMISTRY IMMUNOLOGY MOLECULAR BIOTECHNOLOGY DRUGS VACCINES GENETICS DIAGNOSTICS CHEMICAL ENGINEERING CELL BIOLOGY LIVESTOCK Introduction 14
15 5. Manipulation of gene expression (DSR) Use of expression vector, strategy of overproduction, selection of vectors, selection of host cell. Post test 3 Introduction 15
16 6. Therapeutic T proteins, vaccines and technology to produce recombinant proteins (DSR) Prokaryotic systems, eukaryotic systems (lower and higher eukaryotes), therapeutic proteins and vaccines Quiz 2: topics 4 and 5 STREPTOKINASE W6 W7 W8 F1 F2 F3 M kda INTERFERON 72 kda IFN α2b 33 kda Introduction 16
17 7. Monoclonal antibody and technology to produce monoclonal antibodies (CR) Hibridoma technology and recombinant DNA technology to produce monoclonal antibody, monoclonal antibodies for therapeutic purposes and their mode of actions Post test 4 Introduction 17
18 8. Mid semester exam Introduction 18
19 9. Protein Isolation and Purification (AI) Strategy for protein isolation for intracellular and extracellular proteins, strategies for recombinant protein purification (gel filtration, ion exchange, reverse-phase, affinity, hydrophobic interaction chromatographies) Post test 5 Introduction 19
20 10. Analysis of recombinant products (CR) Protein-based analyses: determination of protein concentration, amino acid sequencing, peptide maping, electrophoresis (SDS-PAGE, native PAGE, electrofocusing, two-dimensional electrophoresis), Western blot. DNA-based analysis: sequencing, hibridization Post test 6 Introduction 20
21 11. Gene therapy and transgenic plants (CR) Explanation, strategy and principles of gene therapy (in vivo, ex vivo), aims of gene therapy, examples of gene therapy, explanation of transgenic plants, principles of construction of transgenic plants, use of transgenic plants, examples transgenic plants. Quiz 3: topics 9, 10 and 11 Written assignment and oral presentation Introduction 21
22 12. Biosimilar products (CR) Definition; regulation; patent situation; first generation biosimilar products: EPO, CSFs, bgh, insulin, hepatitis B vaccine, factor VIII, IFN; development of biosimilar and its requirements Post test 7 Interferon alpha erithropoetin Introduction 22
23 ORAL PRESENTATION (selected group) Introduction 23
24 13. Formulation of recombinant proteins (HR) General considerations of protein and peptide formulation, routes of protein and peptide administration, excipients in protein and peptide formulation, techniques in protein and peptide formulation, quality control of final products erythropoetin somatostatin Introduction 24
25 15. Targeted-drug drug delivery system (HR) Definition of conventional and targeted-drug delivery system, types of targeted-drug delivery system, urgency and aims of drug targetting, ligand, homing device dan homing molecule, design of targeted-drug, mechanisms of drug targeting Introduction 25
26 16. Tissue engineering (TS) Tissue engineering as alternative to organ transplantation, advantages of tissue engineering compared to other therapy, basic principles of tissue engineering, main components of tissue engineering. Introduction 26
27 Grading system Midsem: 35 % Post test: 5 % Quiz: 5 % Participation: 5 % Written assignment: 5 % Oral presentation: 5 % (only for selected groups) Final exam: 40% Introduction 27
28 Class rules After class is commenced, students are not allowed to enter the class No remedial for post test (the mark will be 0) No remedial It is NOT allowed to do copy paste from any kinds of resources including from previous assignments Chatting in any form will be punished according to academic rules No cellular phone during class Cellular phone & E- mail : - DSR: / / debbieretnoningrum@yahoo.com - CR: / c_catur@yahoo.com - AI: / anaindrayati@fa.itb.ac.id Introduction 28
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