proteomics 6 lecture8 - Production of Protein Pharmaceuticals
- 최초 등록일
- 2011.09.01
- 최종 저작일
- 2010.06
- 50페이지/
MS 파워포인트
- 가격 1,000원
소개글
1
목차
Review
Six Step Process
Cell Growth Needs
Prototrophs vs. Auxotrophs
Shake Flask Incubator
Fermentors & Bioreactors
Fermentor Scale Up
Cell Isolation/Harvesting
Protein Isolation & Purification
Cell Disruption Methods
Protein Isolation Methods
Electrophoresis
Isoelectric Point (pI)
IEF Principles
Isoelectric Focusing
Column Chromatography
Affinity Chromatography
Size Exclusion Chrom
Size Exclusion Chromatography (SEC)
Sephadex Structure
Ion Exchange Chromatography (IEC)
Ion Exchange Chromatography
IEC Nomenclature
IEC Groups
IEC Techniques
IEC Rules of Thumb
Conclusion
본문내용
Review
Escherichia coli
Other bacteria
Pichia pastoris
Other yeast
Baculovirus
Animal cell culture
Plants
Sheep/cows/humans
Cell free
Polyhedra
Six Step Process
Isolation of gene of interest
Introduction of gene to expression vector
Transformation into host cells
Growth of cells through fermentation
Isolation & purification of protein
Formulation of protein product
Cell Growth Needs
A sterile carbon, nitrogen, hydrogen and oxygen source (air and H2O) + trace metals (Zn, Fe, Cu, Ca, Mg, Mn)
A sterile energy source (light, sugar, acetate, methanol, ethanol)
A constant (or near constant) temperature above 20 oC
A growth regulating chemical (antibiotic)
Prototrophs vs. Auxotrophs
Protrophic cells (bacteria, plants) can produce all essential amino acids, nucleic acids, carbohydrates and lipids from simple nutrients (water, oxygen, nitrogen or ammonia, CO2)
Auxotrophic cells (yeast, insect cells, mammalian) need vitamins, essential amino acids (His, Cys), sugars, lipids, etc. to grow because they have lost this ability through evolution (bacterial symbiosis)
106 cells/mL
109-1010 cells/mL
Growth density
105 cells/mL
1 cell
Seeding density
10000-100000 nm
100-2000 nm
Size
Very susceptible
Less affected
Environmental FX
Key for buffering
Sometimes
CO2 Requirement
Complex
Usually simple
Nutritional Rqmt
Low
High
O2 Requirement
1-5% per hour
10-50% per hour
Growth Rate
Present
Present
Cell membrane
Generally absent
Generally present
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