Match the following types of column chromatography with the underlying principle. Affinity chromatography HPLC lon-exchange chromatography (EX) Size-exclusion chromatography ✓[Choose] exploits differences in molecular size of individual proteins exploits differences in the size and magnitude of the net charge of individual proteins at specific pis requires a ligand (specific binding partner) to attract and retain a target protein relies on high pressure pumps that force proteins through a column [Choose] [Choose]

Match the following types of column chromatography with the underlying principle. Affinity chromatography HPLC lon-exchange chromatography (EX) Size-exclusion chromatography ✓[Choose] exploits differences in molecular size of individual proteins exploits differences in the size and magnitude of the net charge of individual proteins at specific pis requires a ligand (specific binding partner) to attract and retain a target protein relies on high pressure pumps that force proteins through a column [Choose] [Choose]

Fundamentals Of Analytical Chemistry

9th Edition

ISBN:9781285640686

Author:Skoog

Publisher:Skoog

Chapter33: High-performance Liquid Chromatography

Section: Chapter Questions

Problem 33.1QAP

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The principle of affinity chromatography is based on: a) Charge interactions b) Size exclusion c) Hydrophobic interactions d) Specific binding interactions between molecules
You run the chromatography system with paper as the stationary phase and 70% isopropanol as the mobile phase. You determine the Rf of a polar analyte to be 0.40. If you use 100% isopropanol (a less polar solvent) instead of isopropanol as the mobile phase what would you expect to happen to the Rf value? No value effect on Rf higher lower could be higher or lower
Briefly explain how cation exchange chromatography can used in protein separation?
Which of the following types of liquid chromatography uses immobilized biochemical as a stationary phase? affinity chromatography ion exchange chromatography gel permeation chromatography exclusion chromatography
Which statements are true for molecular exclusion chromatography?I Molecular exclusion chromatography separates on the basis of size.II Small molecules can fit in the pores of the stationary phase and effectively pass through a larger volume, and elute last.III The pores in the stationary phase are too small for large molecules to pass through, so large molecules elute first.IV Molecular exclusion chromatography with a hydrophilic stationary phase and an aqueous solvent is called gel permeation chromatography. A. I, II, III B. III and IV C. II only D. I and II only E. I, III, and IV
Classify the descriptions based on whether they apply to ion‑exchange, size‑exclusion, or both types of chromatography columns. - Proteins move through the column at rates determined by their net charge. - The gel bed will contain polymer beads with charged functional groups. - Proteins move through the column at rates determined by their size. - The gel bed will contain porous polymer beads. - The properties of the proteins affect the rate at which they migrate. - Proteins are separated based on differences in their amino acid sequences.
Which statements are true for molecular exclusion chromatography?I Molecular exclusion chromatography separates on the basis of size.II Small molecules that freely pass through the stationary phase, Kav = 0.III Small molecules can fit in the pores of the stationary phase and effectively pass through a larger volume, and elute last.IV The pores in the stationary phase are too small for large molecules to pass through, so large molecules elute last.V Molecular exclusion chromatography with a hydrophilic stationary phase and an aqueous solvent is called gel permeation chromatography. A. II and V B. I, II, and IV C. III, IV, and V D. No correct combinations given above E. I, III, and IV F. I, II, III, and V
In chromatography, the compound that is least soluble in the mobile phase a) Travels the furthest away from the line of origin b) Stays closest to the line of origin
Which of the following statements in incorrect? O in affinity chromatography a ligand is secured to the column matrix that will associate with a protein of interest O separation of proteins by non-denaturing PAGE will not accurately reflect the size of proteins O lon exchange chromatography only works to separate proteins when the pH of the elution buffer Iis equal to the pl of the proteins O In ion exchange chromatography, the bound proteins are eluted from the column using a high salt concentration solution
"Chromatography in separation is based on the difference in components charge density a O solubility b O migration rate volatility d O
In molecular exclusion (size exclusion) chromatography, the molecules have the longest retention times. a) the larger molecules b) the smaller molecules
The Rf of four spots in a normal phase paper chromatography is given. Determine which of these is the most polar. Rf= 0.40Rf= 0.25Rf= 0.21Rf= 0.34