Column I Column II P. Protein A 1. Size exclusion chromatography Q. Sephadex 2. Ion-exchange chromatography R. Phenylsepharose 3. Affinity chromatography S. Diethylaminoethyl cellulose 4. Hydrophobic interaction chromatography
Chromatography Matching Solution
This question requires matching specific stationary phases (Column I) with their corresponding chromatography techniques (Column II).
Matching Stationary Phases to Techniques
- P. Protein A: Known for binding antibodies specifically. This is characteristic of Affinity chromatography (3).
- Q. Sephadex: A porous polymer used to separate molecules based on size. This is the principle of Size exclusion chromatography (1).
- R. Phenylsepharose: Contains hydrophobic phenyl groups, used for separating molecules based on hydrophobicity, especially under varying salt concentrations. This is Hydrophobic interaction chromatography (4).
- S. Diethylaminoethyl cellulose (DEAE cellulose): A positively charged matrix at typical working pH, used to bind negatively charged molecules. This is Ion-exchange chromatography (2).
Correct Matching Summary
Based on the analysis:
- P matches with 3
- Q matches with 1
- R matches with 4
- S matches with 2
Therefore, the correct combination is P-3, Q-1, R-4, S-2.
Important Questions from Ion Exchange Gel Filtration Hydrophobic Interaction and Affinity Chromatography
- Which of the following separation processes is/are based on molecular size?
- A mixture contains three similarly sized peptides P, Q and R. The peptide P is positively charged, Q is weakly negative and R is strongly negative. If this mixture is passed through an ion-exchange chromatography column containing an anionic resin, their order of elution will be
- Two monomeric His-tagged proteins of identical molecular weight are present in a solution. pIs of these two proteins are 5.6 and 6.8. Which one of the following techniques can be used to separate them?
A protein is to be purified using ion-exchange column chromatography. The relationship between HETP (Height Equivalent to Theoretical Plate) and the linear liquid velocity of mobile phase is given by:
$H = \frac{A}{u} + Bu + C$
where H is HETP (m) and u is linear liquid velocity of mobile phase ($m.s^{-1}$). The values of A, B and C are $3\times10^{-8} \ m^2.s^{-1}$, $3 \ s$ and $6\times10^{-5} \ m$, respectively. The number of theoretical plates based on minimum HETP for a column of 66 cm length will be ____________________.
- Match the entries in the Group I with the elution conditions in Group II.
Group I Group II P. Ion-exchange chromatography 1. Isocratic solvent Q. Hydrophobic column chromatography 2. Ampholytes R. Gel filtration chromatography 3. Increasing gradient of salt S. Chromatofocusing 4. Decreasing gradient of polarity
