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Question 4 please! I posted 3 for help on question 4

Question 4 please! I posted 3 for help on question 4

Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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Question 4 please! I posted 3 for help on question 4
3. A mixture of proteins shown in the table below are dissolved in water and injected into a size exclusion chromatography system. 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 (RI 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 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. Conc 0.5 g/L 1 g/L 12 g/L 5 g/L 2 g/L 10 g/L log(Molecular Weight) 8765432 pl 7.8 6.5 0 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 3log (MW) = 5.262 -0.0378t R² = 0.9991 10 20 Elution Time, min 30
Transcribed Image Text:3. A mixture of proteins shown in the table below are dissolved in water and injected into a size exclusion chromatography system. 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 (RI 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 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. Conc 0.5 g/L 1 g/L 12 g/L 5 g/L 2 g/L 10 g/L log(Molecular Weight) 8765432 pl 7.8 6.5 0 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 3log (MW) = 5.262 -0.0378t R² = 0.9991 10 20 Elution Time, min 30
4. A. Using ultrafiltration as the only separation method (with as many different MWCO membranes as needed from those listed in the class notes on filtration, reproduced below), develop a process to isolate/purify two products from the same mixture in question 3: insulin and lactoperoxidase. (that is come up with two different products that contain the highest concentration of (1) insulin and (2) lactoperoxidase, with minimal other components in these two product streams. Note: % recovery in the table refers to the % of each biochemical recovered in the retentate; the rest passes through the membrane into the permeate, and can be recovered in the stream that crosses the membrane. Dashes in the table indicate that that the particular biochemical was not evaluated in that membrane. Solute B. Without using filtration or size exclusion chromatography, discuss one other technique covered in class that could work if the desired product from this mixture were pure trypsin. Solute MW (Kd) Vitamin B12 Aprotinin Cytochrome C Chymotrypsinogen A Ovalbumin BSA Phosphorylase B IgG Thyroglobulin (1 mg/mL) 1,335 6,200 12,400 25,000 45,000 67,000 97,400 156,000 677,000 MWCO Spin Time (min.) % Recovery % Recovery % Recovery % Recovery % Recovery % Recovery % Recovery % Recovery % Recovery 3K 15 7 99 100 - . 10K 10 - 51 89 97 97 - - 30K 8 - 11 77 94 92 100 95 - - 100K 5 - - 1.8 2.1 3 26 91 97 100 300K 3 - 2. 1.5 1 1.5 91
Transcribed Image Text:4. A. Using ultrafiltration as the only separation method (with as many different MWCO membranes as needed from those listed in the class notes on filtration, reproduced below), develop a process to isolate/purify two products from the same mixture in question 3: insulin and lactoperoxidase. (that is come up with two different products that contain the highest concentration of (1) insulin and (2) lactoperoxidase, with minimal other components in these two product streams. Note: % recovery in the table refers to the % of each biochemical recovered in the retentate; the rest passes through the membrane into the permeate, and can be recovered in the stream that crosses the membrane. Dashes in the table indicate that that the particular biochemical was not evaluated in that membrane. Solute B. Without using filtration or size exclusion chromatography, discuss one other technique covered in class that could work if the desired product from this mixture were pure trypsin. Solute MW (Kd) Vitamin B12 Aprotinin Cytochrome C Chymotrypsinogen A Ovalbumin BSA Phosphorylase B IgG Thyroglobulin (1 mg/mL) 1,335 6,200 12,400 25,000 45,000 67,000 97,400 156,000 677,000 MWCO Spin Time (min.) % Recovery % Recovery % Recovery % Recovery % Recovery % Recovery % Recovery % Recovery % Recovery 3K 15 7 99 100 - . 10K 10 - 51 89 97 97 - - 30K 8 - 11 77 94 92 100 95 - - 100K 5 - - 1.8 2.1 3 26 91 97 100 300K 3 - 2. 1.5 1 1.5 91
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