Chromatography Flashcards
What’s the difference between LPLC and HPLC?
LPLC - no pressure applied, in glass columns
HPLC - high pressure applied, I’m stainless steel columns
What are the different types of chromatography?
Size exclusion chromatography
Ion exchange chromatography
Affinity chromatography
Reversed phase chromatography
In ion exchange chromatography, what are proteins separated based on?
Their net charge
In size exclusion chromatography, what is eluted first?
Largest proteins
In reverse phase chromatography, what is eluted first?
Least hydrophobic proteins
What is special about affinity chromatography?
The only one that created very pure protein fractions in one step
In size exclusion chromatography which beads are used?
Porous beads
In ion exchange chromatography, which proteins elude first?
Negatively charged proteins
How are the beads in the matrix of affinity chromatography linked?
Covalently
In reversed phase chromatography, what does the matrix contain?
Scilia with a chemically attached alkyl group
How does the protein bind to the matrix in reversed phase chromatography?
Through hydrophobic interactions
In reversed phase chromatography, which proteins elude first?
Least hydrophobic
