Say you have a mixture of four proteins with the following properties: Protein A: MW of 30 kDa,...

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Biology

Say you have a mixtureof four proteins with the following properties:
Protein A: MW of 30kDa, pI = 5.0, binds to nitrilotriacetic acid
Protein B: MW of 35kDa, pI = 4.5, low solubility and will precipitate in 5% (w/v)(NH4)2SO4
Protein C: MW of 100kDa, pI = 4.7
Protein D: MW of 30kDa, pI = 7.4

(a) Using a flow chart, propose a purification strategy topurify and separate the proteins
(b) Whattechnique(s) would you use to verify the success of each step ofyour purification?

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ANSWER A and B Isolation of Protein D The first method that should be made applicable is isoelectric focussing of the protein molecules The pH should be kept at 6 and this would result in the separation of two types of proteins namely one which will be positively chared and the others which will be negatively charged The Protein D will be positively charged mainly because it has a pI value which is more shifted towards the basic value as compared to See Answer

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