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Chapter 26: Q18P (page 747)

Would molecular exclusion, affinity, or hydrophobic interaction chromatography be most appropriate for each of the following applications?

  1. Purifying and concentrating a crude mixture of an antibody.
  2. Desalting a solution containing a 30kDa protein.
  3. Finding the molecular mass distribution of polystyrene with 15kDa average mass.
  4. Separation of cytochrome c (12 400 Da) and ribonuclease A (12 600 Da). Cytochrome c has lower surface hydrophobicity than ribonuclease A.

Short Answer

Expert verified

The explanations to the above questions are:

  1. Affinity
  2. Molecular exclusion
  3. Molecular exclusion
  4. Hydrophobic interaction

Step by step solution

01

Explanations to the answers:

  1. Purifying and concentrating a crude mixture of an antibody –Affinity
  2. Desalting a solution containing a 30 kDa protein – Molecular exclusion
  3. Finding the molecular mass distribution of polystyrene with 15 kDa average mass –Molecular exclusion
  4. Separation of cytochrome c (12 400 Da). Cytochrome c has lower surface hydrophobicity than ribonuclease A – Hydrophobic Interaction.
02

Definitions:

  • Affinity chromatography – a method based on macromolecular binding interactions, used for separating biomolecules (like proteins) from a solution.
  • Molecular exclusion – also called Gel Exclusion Chromatography, a method used for separating molecules in a solution based on their size and in sometimes on their molecular weight.
  • Hydrophobic interaction chromatography – a method used for separating molecules based on their hydrophilicity.

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Most popular questions from this chapter

Injection in some homebuilt capillary electrophoresis instruments is performed by raising the sample vial to create a siphon. The pressure exerted by a column of water of heightiswhereis the density of water and gis the acceleration of gravity (9.8m/s2). To what height would you need to raise the sample vial to create the necessary pressure to load the sample in 4.0 s? Is it possible to raise the inlet of this column to this height? How could you obtain the desired pressure?

(a) Suppose that the reservoir in Figure 26-7 contains 1.5 L of 2.0 M K2PO4. For how many hours can the reservoir provide 20mM KOH at a flow rate of 1.0 mL/min if 75% consumption ofin the reservoir is feasible?

(b) What starting and ending current would be required to produce a gradient from 5.0 mM KOH to 0.10 M KOH at 1.0 mL/min flow rate?

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(a) What is the probable charge (Xn-)of pyruvate (peak 10), 2-oxovalerate(peak 16), and maleate (peak 28)? (Hint: Look at the ions around the peaks in question.)

(b) Iodide (peak 44) is a-1ion. Explain its strong retention.

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