Which chromatography test fits best?

In summary, a purified protein fraction containing buffer with salt can be desalted using gel filtration chromatography, paper chromatography, affinity chromatography, or thin layer chromatography. Gel filtration is the best technique for this situation, as it can effectively separate the protein from the salt. Affinity chromatography may also be an option, but it requires the use of chaotropic solvents to remove the protein from the column. Gel filtration can then be used to remove the chaotropic substance.
  • #1
Tyto alba
62
0

Homework Statement



A purified protein fraction containing buffer with salt can be desalted by one of
the following techniques:
(1) Gel filtration chromatography
(3) Paper chromatography
(2) Affinity chromatography
(4) Thin layer chromatography2. The attempt at a solution
Well, the analyte contains salt (possible inorganic so the particles, ions will be small enough as compared to a protein) and protein (a large one) so SEC or gel filtration looks perfect.

But there's affinity chromatography too. Two things come to my mind:
1.having an antibody or enzyme in the stationary phase will definitely work at holding back the proteins
2. but there may be some non specific polar interactions between the protein and the ions, so... (1) 's the right one?
 
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  • #2
With affinity chromatography you will have to use chaotropic solvents to remove the protein from the column. I wouldn't consider this desalting.
 
  • #3
Yes this is easier than your other question, and gel chromatography is the right answer. To elute your bound protein you would have to use either as mentioned chaotropic substance, or perhaps one with specific affinity for your protein. Then you'd probably want to get rid of that - E.g. by gel filtration!
 
Last edited:

1. What is chromatography?

Chromatography is a laboratory technique used to separate and analyze the components of a mixture. It involves passing a mixture through a medium that separates its components based on their different properties, such as size, charge, or polarity.

2. What are the different types of chromatography?

There are several types of chromatography, including gas chromatography, liquid chromatography, ion exchange chromatography, and affinity chromatography. Each type uses a different medium and method of separation.

3. How do I determine which chromatography test is best for my sample?

The best chromatography test depends on the properties of your sample and the components you want to separate. You should consider factors such as the size, charge, and polarity of your sample, as well as the type of medium and detection method that will be most effective.

4. How do I prepare for a chromatography test?

Before conducting a chromatography test, you should carefully select and prepare your sample, choose the appropriate medium and solvent, and calibrate your equipment. It is also important to follow proper safety precautions and record accurate data during the test.

5. What are the applications of chromatography?

Chromatography has a wide range of applications in various fields, including pharmaceuticals, forensics, environmental testing, and chemical analysis. It is commonly used to separate and identify components in mixtures, purify substances, and perform quantitative analysis.

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