Biochemical Engineering CEN 551. Instructor: Dr. Christine Kelly Animal Cell Cultures (Chapter 12) and Glycosylation. Sources. Text - Chapter 12 Peshwa, M. V. Mammalian Cell Culture Websites: http://www.np.edu.sg/~dept-bio/biochemical_engineering/lectures/bioreact1/bioreact3_1.htm.
Instructor: Dr. Christine Kelly
Animal Cell Cultures (Chapter 12) and Glycosylation
(mg/ml)Albumin 35-55Immunoglobulins(IgG 75-85% of all Ig) 8-18Fibrinogen 2-6*Alpha-1 antitrypsin 1-2.5Alpha-2 macroglobulin 0.5-3.5Transferrin 1.5-3.5Alpha-2+ß-lipoproteins (LDL) 4-7Alpha-lipoproteins (HDL) 0.6-1.5Haptoglobin 5Alpha-1 acid glycoprotein 0.5-1.25hemopexin 1Pre-albumin 0.3-0.4Total Protein 62-80
Tissue culture flasks
(lactic acid and ammonium.
Aeration and agitation in mammalian cell culture(following material from http://www.np.edu.sg/~dept-bio/biochemical_engineering/lectures/bioreact1/bioreact3_1.htm)
Vortices form in the wake of the rising bubble.
Flow lines move fastest near the bubble.
Bubble damage rather than liquid shear forces are the major cause of cell damage in sparged animal cell bioreactors
The extent of damage caused by these factors is dependent upon the…
Airlift reactors have been used to to successfully cultivate mammalian and insect cells in reactors with liquid volumes of up to 1000 L. This is despite the potential problems associated with bubble damage.The company Celltech, for example uses airlift production as the predominant technique for large scale cultivation of hybridoma cells.The low shear environment provided by airlift reactors, combined with the use of appropriate media and shear protectorants can compensate for the increased likelihood of bubble damage.
Three main techniques by which enhanced oxygen transfer rates can be achieved without the need for sparging:
Various techniques are used to achieve direct bubble free oxygenation of animal cell bioreactors including the use of gas permeable
Hollow fiber reactors
Immobilization in gel beads
Whole cells and tissue culture
Asn – X – Ser or Thr
Anchor glycosylation directly bound directly to the cell membrane.