Microsoft PowerPoint - 1 Introduction

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1 Pharmaceutical Biotechnology Introduction By Yuqiong Xia 3/2/2016

2 What is pharmaceutical biotechnology Biotechnology use microorganisms, plants, and animals or parts to produce useful compounds Pharmaceutical biotechnology biotechnological manufacturing of pharmaceutical products 2

3 What are biopharmaceuticals Protein and peptides Anti-sense Oligonucleotides (oligos) DNA 3

4 Biopharmaceuticals in market Epoetin-α( 红细胞生成素 )(Epogen, Eprex, Procrit ) Abciximab ( 阿昔单抗 )(ReoPro ) Interferons-α( 干扰素 Intron A, Roferon A), interferons -β (Avonex, Rebif, Betaseron ) Anti-TNF-α agents (Enbrel, Remicade, Humira ) Trastuzumab ( 曲妥单抗 )(Herceptin ) 4

5 What we will learn Part I Basics Molecular biotechnology Protein analysis Protein production and purification Formulation Pharmacokinetics and pharmacodynamics Protein immunogenicity Genomics 5

6 What we will learn Part II Biopharmaceutical drugs Gene therapy Oligonucleotides Hematopoietic Growth Factors Interferons and Interleukins Insulin and Growth Hormones Monoclonal Antibodies 6

7 Reference 7

8 How to get your score 60% final exam 20% literature report 10% homework 10% attendance 8

9 Pharmaceutical Biotechnology Molecular Biotechnology

10 Outline Basics Prokaryotic cell Gene expression Techniques Recombinant DNA technology Special DNA technology Cell cultures 10

11 The Prokaryotic Cell 11

to thick peptidoglycan Gram negative bacteria Cannot retain

12 西安电子科技大学 Gram positive (G + ) and Gram negative (G - 夏玉琼 ) bacteria Gram positive bacteria Can retain crystal violet dye due to thick peptidoglycan Gram negative bacteria Cannot retain crystal violet dye, are colored red or pink 12

13 G + and G - bacteria G + bacteria PG of multilayer G - bacteria PG of Mono or double layer Without LPS Containing toxic LPS OM: Outer Membrane IM: Inner Membrane PG: peptidoglycan LPS: lipopolysaccharide 13

14 Plasmids Small and circular double-stranded DNA molecules Replication Integration 14

15 Outline Basics Prokaryotic cell Gene expression Techniques Recombinant DNA technology Special DNA technology Cell cultures 15

16 central dogma 16

17 DNA replication A. Semi conservative replication 17 B. Closed circle replication C. Rolling cycle replication

18 Transcription mrna, specify the amino acid composition of the cellular proteins trna or rrna, participate as auxiliary molecules for translation 18 mirna, regulate protein synthesis

19 Translation Amino acid Anti-codon Codon 19

20 Outline Basics Prokaryotic cell Gene expression Techniques Recombinant DNA technology Special DNA technology Cell cultures 20

21 21 Recombinant DNA technology/dna cloning

22 DNA transfer by electroporation 22

23 DNA transfer by phage 23

24 西安电子科技大学夏玉琼 DNA transfer by conjugative plasmid( 接合型质粒 ) 24

25 DNA transfer by virus virus infection 25

26 DNA transfer into nucleus Only for animal and plant cells 26

27 DNA transfer by special conjugation Ti: tumor inducing 27 根癌土壤杆菌 Infected plant cells develop crown gall 冠瘿病 Modified Agrobacterium can transfer DNA biological kingdom barriers are crossed in a natural fashion

28 DNA sources Synthetic DNA organic DNA synthesis approach only for small proteins cdna (the copy DNA) From mrna DNA libraries 28

29 夏玉琼 Production by recombinant DNA technology Human insulin 西安电子科技大学 Human growth hormone (hgh)

30 夏玉琼 Production of insulin by recombinant DNA tech 西安电子科技大学 β-gal: β- 半乳糖苷酶, 乳糖启动子的产物 30 Cyanogen bromide: 溴化氰, 水解含有 Met( 甲硫氨酸 ) 的多肽

31 夏玉琼 Production of hgh by recombinant DNA tech 西安电子科技大学 Add a leading DNA Add a part of missing DNA 31 Protein between IM and OM

32 Outline Basics Prokaryotic cell Gene expression Techniques Recombinant DNA technology Special DNA technology Cell cultures 32

33 Special DNA techniques Gene sequencing DNA hybridization PCR technology 33

34 Gene sequencing by Sanger method 34

35 Gene sequencing by Sanger method Primer Fragment to be sequenced Fix reaction mixtures, each contains a ddntp Template C G A T Primer DNA polymerase da(t,c,g)tp ddctp Gel electrophoresis 35

36 DNA hybridization to detect mutant DNA from white blood cells or amnion fluid ( 羊水 ) Boil DNA to make it single-stranded Hybridize with probes probe Hybridized DNA Nonhybridized DNA 36 Modified from

37 PCR technology 37

38 PCR technology 38

Application of PCR technology Detect microbial pathogens

Detect bacterial and viral pathogens in clinical

com http://static.ddmcdn.com/gif/blood-cells.

39 Application of PCR technology Detect microbial pathogens in food Determine the origin of DNA in forensic science Detect bacterial and viral pathogens in clinical material

40 Outline Basics Prokaryotic cell Gene expression Techniques Recombinant DNA technology Special DNA technology Cell cultures 40

bioproducts 指数期 41 迟缓期 http://en.wikipedia.

41 Cultivation of microbes Microbes Medium: nutrient broth, nutrient agar 稳定期衰亡期 Harvest bioproducts 指数期 41 迟缓期 Bacteria growth curve

42 Animal Cell Cultures Cell subculture: PBS, trypsin, culture medium (MEM, DMEM, RPMI 1640)

43 Plant Cell Cultures The target protein only exists in some tissues. Fully differentiated plant tissues are excised to initiate a cell culture, but this differentiated state is usually lost. Add precursor in plant cells, cells act as biosynthetic reactors 43

44 Summary Basics The difference between Gram+ and Gram- bacteria The properties of plasmid Technologies Recombinant DNA technology: DNA transfer DNA sequencing, hybridization, PCR Cell culture Bacteria growth curve Cell medium 44

Microsoft PowerPoint - 1 Introduction.pptx

Microsoft PowerPoint - 1 Introduction.pptx Pharmaceutical Biotechnology Introduction By Yuqiong Xia 3/5/2017 Outline Pharmaceutical biotechnology History of biopharmaceuticals Biopharmaceuticals: current status and future prospects Traditional