Pharmaceutical Biotechnology - Xidianweb.xidian.edu.cn/yqxia/files/20140222_213050.pdfWhat is pharmaceutical biotechnology 2 Biotechnology use microorganisms, plants, and animals or

By Yuqiong Xia2014

Introduction

Pharmaceutical Biotechnology

What is pharmaceutical biotechnology

2

Biotechnology use microorganisms, plants, and animals or parts to

produce useful compounds

Pharmaceutical biotechnology biotechnological manufacturing of pharmaceutical

products

生物制药工程夏玉琼西安电子科技大学

What are biopharmaceuticals

3

Protein and peptides Anti-sense Oligonucleotides DNA


Biopharmaceuticals in market

4

Epoetin-α（红细胞生成素）(Epogen®, Eprex®, Procrit®)

Abciximab （阿昔单抗）(ReoPro®) Interferons-α(Intron®A, Roferon®A), interferons -β

(Avonex®, Rebif®, Betaseron®) Anti-TNF-α agents (Enbrel®, Remicade®, Humira®) Trastuzumab （曲妥单抗）(Herceptin®)


What we will learn

5

Part I Basics Molecular biotechnology Protein analysis Protein production and purification Formulation Pharmacokinetics and pharmacodynamics Protein immunogenicity Genomics


What we will learn

6

Part II Biopharmaceutical drugs Gene therapy Oligonucleotides Hematopoietic Growth Factors Interferons and Interleukins Insulin and Growth Hormones Monoclonal Antibodies


7

Reference

http://web.xidian.edu.cn/yqxia/teach.html


How to get your score

8

60% final exam 20% literature report 10% homework 10% attendance


Molecular Biotechnology

Pharmaceutical Biotechnology 生物制药工程夏玉琼西安电子科技大学

Outline

10

Basics Cell Gene expression

Techniques Recombinant DNA technology Special DNA technology Cell cultures


11

The Prokaryotic Cell生物制药工程夏玉琼西安电子科技大学

G+ cell and G- cell

12

G+ cell G- cell

PG of multilayer PG of Mono or double layer

Without LPS Containing toxic LPSOM: Outer Membrane IM: Inner MembranePG: peptidoglycan LPS: lipopolysaccharide


13

Plasmids

http://en.wikipedia.org/wiki/File:Plasmid_replication_(english).svg


14

The Eukaryotic Cell生物制药工程夏玉琼西安电子科技大学

Outline

15




DNA replication

16


17

Transcription生物制药工程夏玉琼西安电子科技大学

18

RNA splicing生物制药工程夏玉琼西安电子科技大学

19

Translation生物制药工程夏玉琼西安电子科技大学

Outline

20




21

Recombinant DNA technology/DNA cloning生物制药工程夏玉琼西安电子科技大学

22

DNA transfer by electroporation

http://www.btxonline.com/product_images/uploaded_images/electroporation-cartoon.png


23

DNA transfer by phage生物制药工程夏玉琼西安电子科技大学

24

DNA transfer by conjugative plasmid

http://en.wikipedia.org/wiki/File:Conjugation.svg


25

DNA transfer by virusvirus infection


DNA transfer into nucleus

26

Only for animal and plant cells


DNA transfer by special conjugation

27

Infected plant cells develop crown gall 冠瘿病 Modified Agrobacterium can transfer DNA biological kingdom barriers are crossed in a natural fashion

根癌土壤杆菌

Ti:�tumor�inducing


DNA sources

28

Synthetic DNA organic DNA synthesis approach only for small proteins

cDNA (the copy DNA) From mRNA

DNA libraries


Production by recombinant DNA technology

29

Human insulin

Human growth hormone (hGH)

http://t0.gstatic.com/images?q=tbn:ANd9GcTyGER40Hz8bzW81RxVH-GeTdy4fUbZY9LiNi6K7VYSVNABmkwDhttp://gene-vinate.com/doc_images/hegf_image1.jpg


30

Production of insulin by recombinant DNA tech

β-gal: β-半乳糖苷酶，乳糖启动子的产物

Cyanogen bromide：溴化氰，水解含有Met(甲硫氨酸)的多肽


31

Production of hGH by recombinant DNA tech

http://www.uic.edu/classes/bios/bios100/lectures/recomb01.jpg

Add a part of missing DNA

Add a leading DNA

Protein between IM and OM


Outline

32




Special DNA techniques

33

Gene sequencing DNA hybridization PCR technology


Gene sequencing by Sanger method

34 http://www.uic.edu/classes/bios/bios100/lectures/ddNTP.jpg


Gene sequencing by Sanger method

35http://www.scq.ubc.ca/genome-projects-uncovering-the-blueprints-of-biology/

C G A T

Primer Fragment to be sequenced

Fix reaction mixtures,each contains a ddNTP

Gel electrophoresis

TemplatePrimerDNA polymerasedA(T,C,G)TPddCTP


36

DNA hybridization to detect mutant

Modified from http://classes.midlandstech.edu/carterp/Courses/bio225/chap10/ss3.htm

DNA from white blood cells or amnion fluid (羊水)

Boil DNA to make it single-stranded

Hybridize with probes probe

Hybridized DNA

NonhybridizedDNA


37

PCR technology

https://www.boundless.com/image/the-process-of-pcr-in-dna-amplification--2/


38

Application of PCR technology

https://encrypted-tbn2.gstatic.comhttp://static.ddmcdn.com/gif/blood-cells.jpghttp://www.collegecrunch.org/wp-content/uploads/2009/09/forensic-science-350x350.jpg

Detect microbial pathogens in food

Detect bacterial and viral pathogens in clinical material

Determine the origin of DNAin forensic science


Outline

39




40

Cultivation of microbesMicrobesGrowth medium: nutrients

http://en.wikipedia.org/wiki/File:Anthrax_culture.jpg

Harvest bioproducts

Bacteria growth curve


41

Animal Cell Cultures

http://www.zet.or.at/spool/gfx/1201165054-SerumfreeDB.jpghttp://en.wikipedia.org/wiki/Cell_culture

Need all nutrients, special growth factors and hormones


42

Plant Cell Cultures

http://www.ime.fraunhofer.de/en/businessareasMB/integrated_production_platforms/plant_cell_culture.html

can not be kept in the differentiated state, but the target protein only exists in some tissues

Add precursor in plant cells


Summary

43

Basics The difference between Gram+ and Gram- bacteria The properties of plasmid

Technologies Recombinant DNA technology: DNA transfer DNA hybridization

Cell culture Bacteria growth curve


Summary of keywords

44

Prokaryotic cell, eukaryotic cell Bacteria, microbe, virus, plasmids Cytoplasm, cell envelop Transcription, translation, “central dogma” Recombinant DNA technology


Documents

Pharmaceutical Biotechnology - Xidianweb.xidian.edu.cn/yqxia/files/20140222_213050.pdfWhat is pharmaceutical biotechnology 2 Biotechnology use microorganisms, plants, and animals or