3. (10 pts) A mixture of proteins shown in the table below are dissolved in water and injected into a size exclusion chromatograph. Given the calibration curve for the SEC column, draw a qualitative curve to show the expected results (chromatogram) of running the SEC by showing detection units (Rl or UV absorbance) on the y-axis and time on the y-axis. Relative peak heights (areas under curves) and times should be as quantitative as possible. ID 1 2 3 4 5 6 Protein Collagen Lactoperoxidase Insulin Fibronectin Lysozyme Trypsin Molecular Weight 68,000 92,600 5,800 55,000 16,000 47,000 Conc 0.5 g/L 1 g/L 12 g/L 5 g/L 2 g/L 10 g/L pl 7.8 6.5 7.3 4.6 4.6 2.5 Type fibrous a-helices; structural antibacterial enzyme globular protein fibrous protein cell-lysing enzyme digestive enzyme Calibration Curve for SEC Column 8 Note: as shown in the calibration curve, the beads in the column can separate molecules less than ~120,000 molecular weight, but any proteins larger than that will elute at ~5 minutes. The minimum molecular weight that can be separated in the column is is 20,500; smaller proteins elute at 25 minutes. The equation of the calibration curve is shown on the graph. log(Molecular Weight) Osn m NO 3log (MW) = 5.262 - 0.0378t R2 = 0.9991 0 O 10 20 30 Elution Time, min