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Cell disruption
-Key technique in DSP to release intracellular product.
Biological products:
1. Extracellular
2. Intracellular
3. Periplasmic
Cells
• Gram positive bacterial cells
• Gram negative bacterial cells
• Yeast cell
• Mould cells
• Cultured mammalian cells
• Cultured plant cells
• Tissue
What are we disrupting?
Yeast and mould
Physical methods
• Disruption in bead mill
• Disruption using a colloid mill
• Disruption using French press
• Disruption using ultrasonic vibrations
Cascading
beads
Rolling
beads Cells being
disrupted
Cell Stator
suspension
Plunger
Cell Cylinder
suspension
Jet
Orifice
Impact plate
Ultrasound tip
Cell suspension
π = RT (ci − co )
•R = gas constant
T = absolute temperature (K)
ci-co = difference between total solute molarity
inside and outside the cell
Chemical and physicochemical
methods
•Detergents
•Enzymes
•Organic solvents
•Osmotic shock
•Alkali treatment
Choice of Disruption Method
• The method selected for large scale cell
disruption will be different in every case,
but will depend on:
– Susceptibility of cells to disruption
– Product stability
– Ease of from cell debris
– Speed of method
– Cost of method
Adverse Factors during Disruption
• Heat
• Shear
• Proteases
• Particle size
• DNA, RNA
• Chemical
• Foaming
• Heavy-metal toxicity
Thank you