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If a protein with pi of 6 is purified on a DEAE column is it better to use pH 4ph 6 of pH 8 why?
Wiki User
∙ 14y ago
DEAE columns contain a positively charged resin to which negatively charged molecules and proteins will bind. In protein purification, one will generally have the target protein bind the column so the non-target proteins will "flow through" after which the bound proteins are "washed off" the column by changing either the pH or salt concentration. Since the pI of the protein is 6.0, at pH=6.0, the protein has a net charge of zero and will not bind the column, so this pH is not suggested. When the pH is greater than the pI, the protein has a positive charge, so at pH=8.0 the protein will be repulsed by the positively charged resin and will not bind, so pH=8.0 is also not recommended. When the pH is less than the pI, the protein carries a negative charge and will bind the DEAE column and can thus be purified, so the pH=4.0 condition will be productive toward protein purification for this protein.
Wiki User
∙ 14y ago
It would be better to use a pH of 4 for purification on a DEAE column. This is because at pH 4 (below the protein's pI of 6), the protein will have a net positive charge and will bind strongly to the negatively charged DEAE resin, allowing for effective purification.
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