Two hFGF-1 mutants, A and B, are purified using a heparin-Sepharose column. Mutant A is eluted...

Two hFGF-1 mutants, A and B, are purified using a heparin-Sepharose column. Mutant A is eluted upon addition of 0.1M NaCl. Mutant B is eluted upon addition of 0.2 M NaCl. Which protein has a higher affinity for heparin? Is either protein expected to be 100% pure after this purification step?

Expert Solution

A = 0.1 M

B = 0.2 M

higher affinity to NaCl will be the one with least concentration, since it elutes faster and "easier" on it

so

A elutes better on Nacl (ionic) solution than B

therefore

B has much more affinity toward the column,

The 100% separation is not likely to occur, since in chromatography or elution we have just partial separations, due to the coefficients of distributino in polar/ionic solvents

queen_honey_blossom answered 3 years ago

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