Depicted below are one bead with a pore, and two molecules with different sizes.
- What will happen as each molecule interacts with the pore?
- If a column is packed with these porous beads, shown as below, how would this effect the movement of both molecules through the column?
- What will happen to an intermediate sized molecule?
- Draw a separation chromatogram if a group of these three molecules go through this column.
- What contribute to the total volume of the column?
- Can we construct a calibration curve for this type of separation? How?
Hint: molecular size is directly related to the formula mass of the molecules.
- If the pore sizes change, what would happen to the calibration curve? Do these curves make sense?
- What type of applications of size exclusion chromatography can you think of?
- Which is more difficult to separate, glucose and immunoglobulin, or two immunoglobulins? Why?