1. ERT314 BIOREACTOR SYSTEM CHAPTER 2: BIOLOGICAL SYSTEMS AND MEDIA DESIGN

2. CHAPTER 2: BIOLOGICAL SYSTEMS AND MEDIA DESIGN Introduction to Biological System Design, Formulation & Optimization of Media

3. 2.1 Introduction to Biological System

12. ** rheology – the study of the flow of matters, especially in liquid state

15. Fermentation with fungiFermentation with bacteria

19. Oxygen Demand  OTR = Oxygen transfer rate  OUR = Oxygen uptake rate

23.  Most organisms reduce the pH during growth.

33. CHAPTER 2: BIOLOGICAL SYSTEMS AND MEDIA DESIGN Introduction to Biological System Design, Formulation & Optimization of Media Microbial process Plant cell Animal / mammalian cell LET’S RECALL…

36. e.g: glycoprotein, oligosaccharides, enzyme etc. E.Coli has a system where the capacity to use these precursors exceed the capasity of intermediary metabolism

37. 2.2.1 Microbial Processes A. Design and Formulation of Media Should consider:

38. e.g: e.g: antagonism (Zn + to Cu 2+ ), synergism (toxic Cu 2+ + Cd 2+ ), stimulation (Ca 2+ and Mn 2+ depends on Zn 2+ )

39. requires higher pantothenic acid content in media Addition of glycine betaine in media for L- amino acid production which has high glucose concentration, acts as an osmotic protectant.

41. (if more than 10 mM, no antibiotic is synthesized)

42. (easily altered)

43. causing Whereas others, like dried yeast are generally uniform from batch to batch.

44. General equation for the stoichiometry of growth and product formation

45. Where y x/s, y p/s and y CO2/s are = coefficients related to biomasss, product and CO2 formation = expressed as C mole of biomass, product and CO2 per mole of carbon and energy source (substrate) σ i = carbon fraction of i (x,p,s) Y x/s = g dry weight of biomass per g of substrate consumed Y p/s = g dry weight of product per g of substrate consumed

49. Constraints:  Low reactor rated and productivity  Instability of cell lines presents problem not easy to overcome  Danger of microbial contamination  Secondary metabolites is often retained in biomass  Plant cells are often shear sensitive  In some cases, secondary compounds should be removed rapidly How to overcome?

65. ,

66. Some of the important products which are produced from animal cell cultures are : (i)enzymes (asperagenase, collagenase, urokinase, pepsin, hyaluronidase, rennin, trypsm, tyrosin hydroxylase), (ii) hormones (leutinizing hormone, follicle stimulating hormone, chorionic hormone and erythropoietin), (iii) vaccines (foot and mouth disease vaccine, vaccines for influenza, measles and mumps, rubella and rabies), (iv) monoclonal antibodies etc.

67. ProductsApplication Erythropoietins Erythropoietin-aAnaemia resulting from cancer and chemothcraoy Erythropoietin-pAnaemia secondary to kindney disease Human growth hormones hGHHuman growth deficiency in children, renal cell carcinoma SomatotropinChronic renal insufficiency, Turners' syndrom Monoclonal antibodies (therapeutic) Anti-lipopolysacharideTreatment of sepsis Murine anti-idiotype/human B-cell lymphoma Monoclonal antibodies (diagnostics) Anti-fibrin 99Blood clot 99 Tcm-FAb (breast)Blood cancer PR-356CYT-356-in-lllProstate adinocarcinoma Plasminogen activator Urokinase type plasminogen activator Acute myocardial infarction, acute stroke, pulmonary embolism, deep vein thrombosis Tissue type plasminogen activator Recombinant plasminogen activator Vaccines HIV vaccines (gpl20)AIDS prophylaxis and treatment Malaria vaccineMalaria prophylaxis Polio vaccinesPoliomyelitis prophylaxis Some products of medical use derived from animal cell cultures. Source: based on Feichter (1996).

68.  Advantages of serum in culture medium are: i) serum binds and neutralizes toxins, (ii) serum contains a complete set of essential growth factors, hormones, attachment and spreading factors, binding and transport proteins, (iii) it contains the protease inhibitors, (iv) it increases the buffering capacity, (v) it provides trace elements.

69. Disadvantages of serum in culture medium are: (i) it is not chemically defined and therefore it’s composition varies a lot, (ii) it is sometimes source of contamination by viruses etc. (iii) it increases the difficulties and cost of down stream processing, (iv) it is the most expensive component of the culture medium. THEREFORE,

75. (1) Immunisation of a mouse (2) Isolation of B cells from the spleen (3) Cultivation of myeloma cells (4) Fusion of myeloma and B cells (5) Separation of cell lines (6) Screening of suitable cell lines (7)(a) in vitro or (b) in vivo multiplication (8) Harvesting Production of monoclonal antibody (briefly)

78. THANK YOU