During purification, what happens to hydrophobic proteins bound to a hydrophobic resin when eluted?

During purification, what happens to hydrophobic proteins bound to a hydrophobic resin when eluted?

• A. They are released from the resin during elution.
• B. They remain bound permanently.
• C. They are degraded.
• D. They are cleaved from the protein.

Answer: (A)

During elution, the conditions are changed to weaken the hydrophobic interactions that hold the protein to the hydrophobic resin. In this type of chromatography, high salt strengthens those interactions, so binding occurs; as the salt concentration is lowered or the mobile phase becomes more polar, the hydrophobic forces diminish and the protein desorbs from the resin. So, the proteins bound to the hydrophobic resin are released into the eluate during elution, rather than remaining attached, being degraded, or being cleaved.

During elution, the conditions are changed to weaken the hydrophobic interactions that hold the protein to the hydrophobic resin. In this type of chromatography, high salt strengthens those interactions, so binding occurs; as the salt concentration is lowered or the mobile phase becomes more polar, the hydrophobic forces diminish and the protein desorbs from the resin. So, the proteins bound to the hydrophobic resin are released into the eluate during elution, rather than remaining attached, being degraded, or being cleaved.