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Dialysis and Filtration

Quiz 1. Describe the curve for dialysis over time Describe what controls the rate of mass transfer in dialysis. Show by calculation the number of buffer changes to reach 1 millimolar salt concentration from an initial concentration of 1 molar if you have 100 ml of solution in a dialysis bag and a

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Dialysis and Filtration

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    1. Dialysis and Filtration Dealing with the little things

    2. Quiz 1 Describe the curve for dialysis over time Describe what controls the rate of mass transfer in dialysis. Show by calculation the number of buffer changes to reach 1 millimolar salt concentration from an initial concentration of 1 molar if you have 100 ml of solution in a dialysis bag and a 500 ml beaker. State any assumptions you make about the system. Assume there is an infinite supply of salt free buffer.

    3. Parts             1.         Theory             2.         Dialysis tubing/cassettes             3.         Filters and membranes             4.         Ultrafiltration             5.         Microconcentrator

    4. Dialysis Permeable membrane that selects molecules based on size. Lower end of molecular weight cut off is about 8-10kd Standard tubing is 12-14 kd Rate of flux is jd = K p,dialysis DC J= flux K = permeability constant of dialysis, DC=Concentration difference

    6. Ultra-filtration utilizes permeable membrane filters to separate the components of solutions and suspensions based on their molecular size used for the concentration, desalting, or fractionation of mixtures of biomolecules. Membranes come in a variaty of pore sizes from as small as 1000 to 1,000,000 da > Millipore and Pall are the biguns

    8. Ultra-filtration Concentration Choose a membrane that retains the molecule of interest desalting/buffer exchange Choose a membrane that retains the molecule of interest and lets ligands and salts through. fractionation of mixtures of biomolecules One can either retain and concentrate Pass through

    9. Ultra-filtration

    10. Ultra-filtration

    11. Ultra-filtration Several different styles of units Centrifugal systems Uses centrifugal force to generate pressure gradient Direct VS indirect

    12. Millipore* Microcon*, Centricon* and Centriplus

    13. Millipore* Centriprep*

    14. Millipore* Ultrafree

    15. Continuous filtration

    16. Ultra-filtration kinetics

    17. Buffer exchange rates

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