Ion exchange chromatography

Ion exchange chromatography: a brief account use full for industrial microbiology paper

1. ION EXCHANGE
CHROMATOGRAPHY
Dr. Deepak Bhusare
Department of Microbiology
DB, ACS College, Bhokar.

2. Ion Exchange Chromatography
Ionexchangechromatography--isaseparationbasedoncharge.
Usedforalmostanykindofchargedmolecules---largeproteins,
smallnucleotidesandaminoacids.
Mobilephage–buffer,pHandsaltconcentration---opposite
chargedsoluteionsattractedtothestationaryphagebyelectrostatic
force.
Stationaryphage–resinisusedtocovalentlyattachanionsor
cationsontoit

3. Principle……….
IonExchangeChromatographyreliesoncharge-charge
interactionsbetweentheproteins.
Ionexchangechromatography,whichisalsoknownas
adsorptionchromatography.

4. Types of IEC….
Anionexchangers
Cationexchangers
•Exchanger types:
1.Weak cations
2.Strong cations
3.Weak anions
4.Strong anions

5. Cation = Positive charge
Anion = Negative charge

6. Cation exchange chromatography
Positivelychargedmoleculesareattractedtoanegatively
chargedsolidsupport.
CommonlyusedcationexchangeresinsareS-resin,sulfate
derivatives;andCMresins,carboxylatederivedions

7. Anionexchangechromatography
Negativelychargedmoleculesisattractedtoapositively
chargedsolidsupport.
•CommonlyusedanionexchangeresinsareQ-resin,a
Quaternaryamine;andDEAEresin,DiEthylAminoEthane

8. Several matrix types and their important properties
Cellulose;Hydrophilicsurface,enhancedstabilitybycross-linking,
inexpensive.
Dextran;Considerableswellingasafunctionofionicmilieu,
improvedmaterialsbycrosslinking).
Agarose;SwellingisalmostindependentofionicstrengthandpH,
highbindingcapacityobtainedbyproductionofhighlyporous
particles.
Polyacrylamide;Swellingbehaviorsimilartodextran.
Acrylate-copolymer;HighpHstability.
Polystyrene-divinilybenzene;Hydrophobicsurface,lowbinding
capacityforproteins.
Silica;UnstableatpH>8,rigidparticles.

10. Chromatography Methods
ColumnwashedwithbufferAtoequilibrate
BufferBisusedtoequilibrateagain
EquilibratethecolumnwithbufferA
Sampleloading
Flowthroughcollection
Eluteprotein

11. Advantages
HighCapacity
HighResolvingPower
MildSeparationConditions.
VersatilityAndWidespearedApplicability
TendencyToConcentrateTheSample
RelativelyLowCost

12. Disadvantages
Costlyequipmentandmoreexpensivechemicals
Turbidityshouldbebelow10ppm.

13. Conclusion
Ionexchangechromatographyismoreefficientthanother
chromatography.Itcouldbewidelyusedforcommercial
purposes.
Ref:www.google.com

14. Thank you…