14. A protein mixture consisting of proteins A, B, and C was subjected to various protein purification steps. First, the protein mixture was applied to an anion ion exchange column. When the column was washed with an increasing concentration gradient of chloride ions, the order of elution of the three proteins was B, then C, and finally A. Next, the protein mixture was chromatographed on a gel filtration column. The order of elution of the three proteins from this column was C, A, B. Finally, the protein mixture was passed over a chromatography column containing a Ni-NTA affinity matrix. Only protein A was retained by the column. Answer the following (i) Which protein (A, B, or C) has the highest and lowest positive charge (ii) Which protein has the highest and lowest mass (ii) Which protein has an affinity for the Ni-NTA matrix and why

Biochemistry
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Chapter1: Biochemistry: An Evolving Science
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14. A protein mixture consisting of proteins A, B, and C was subjected to various protein purification
steps.
First, the protein mixture was applied to an anion ion exchange column. When the column was
washed with an increasing concentration gradient of chloride ions, the order of elution of the three
proteins was B, then C, and finally A.
Next, the protein mixture was chromatographed on a gel filtration column. The order of elution of the
three proteins from this column was C, A, B.
Finally, the protein mixture was passed over a chromatography column containing a Ni-NTA affinity
matrix. Only protein A was retained by the column.
Answer the following
(i) Which protein (A, B, or C) has the highest and lowest positive charge
(ii) Which protein has the highest and lowest mass
(ii) Which protein has an affinity for the Ni-NTA matrix and why
Transcribed Image Text:14. A protein mixture consisting of proteins A, B, and C was subjected to various protein purification steps. First, the protein mixture was applied to an anion ion exchange column. When the column was washed with an increasing concentration gradient of chloride ions, the order of elution of the three proteins was B, then C, and finally A. Next, the protein mixture was chromatographed on a gel filtration column. The order of elution of the three proteins from this column was C, A, B. Finally, the protein mixture was passed over a chromatography column containing a Ni-NTA affinity matrix. Only protein A was retained by the column. Answer the following (i) Which protein (A, B, or C) has the highest and lowest positive charge (ii) Which protein has the highest and lowest mass (ii) Which protein has an affinity for the Ni-NTA matrix and why
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