On souhaite évaluer la pureté d'un anticorps monoclonal (bevacizumab) utilisé à des fins thérapeutiques pour traiter le cancer du sein. Séquence d'acides aminés - Masse molaire: ~150 kDa Quelles précautions dois-je prendre lorsque je choisis ma colonne chromatographique? Quelle diamètre de colonne et quelle chimie de phase stationnaire utiliseriez-vous?
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For monoclonal antibodies like bevacizumab, size-exclusion chromatography (SEC) or ion-exchange chromatography (IEX) are commonly used. Show more…
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Precipitation of monoclonal antibody: Human IgM monoclonal antibody is produced by hybridoma cells in a stirred fermenter. Antibody secreted into the culture medium reaches a concentration of 120 μg/ml of cell-free broth. The product is concentrated by precipitation immediately after removal of the cells. The yield of antibody recovered in the precipitate is determined as a percentage of the initial mass of antibody present in the broth. The precipitate purity, or the percentage of the precipitate weight that is IgM antibody, is also measured. Various precipitation methods are tested and the following results are obtained. From the data provided, what are the advantages and disadvantages of using 12% w/v polyethylene glycol for this precipitation process compared with 8% w/v? (b) Using % w/v as the unit for salt concentration and μg/g of cell-free broth as the unit for protein solubility, derive an empirical equation for IgM solubility as a function of ammonium sulfate concentration at pH 7.2. The density of cell-free broth can be taken as 1000 kg/m^3. (c) Predict the antibody yield using 27% w/v (NH4)2SO4. (d) What mass of antibody will remain in solution if 27% w/v (NH4)2SO4 is used to treat 100 L of cell-free broth?
Lainey R.
PART 1: Analysis of Urine to Detect Disease Title: Examination of Urine for Disease Detection Introduction: The excretory system plays a crucial role in maintaining homeostasis in the body. The kidneys, as a vital component of this system, filter waste products from the blood and produce urine. Urinalysis, the analysis of urine, provides valuable insights into the overall health of an individual. This report aims to investigate the structure and function of the kidney, as well as the process of urinalysis. Additionally, it will explore the detection of biomolecules in urine and their significance in diagnosing diseases. The experiment conducted in this study involves testing urine samples for glucose, protein, and bile. The presence or absence of these substances can indicate various health conditions. By understanding the biological explanations behind abnormal biomolecules found in urine, potential diagnoses for each patient can be determined. Furthermore, the reliability and accuracy of the experiment will be evaluated, along with suggestions for improvement. The benefits and limitations of using urine tests for disease diagnosis will also be discussed. Method: The urine samples were tested for glucose, protein, and bile using the following steps: [Provide a clear description of the steps involved, including the equipment used, in an academic style (past tense and passive voice).] Recording the result: The data obtained from the tests should be arranged in a suitable table, including the test outcomes and inferences drawn from them. Discussion: A brief summary of the results will be provided, along with possible diagnoses for each patient. The biological explanations for the presence of abnormal biomolecules in the urine will be explained, considering the structure and function of the kidney and other organs. Evaluation of the experiment: The process of urinalysis will be assessed, focusing on the reliability and accuracy of the experiment. Suggestions for improving the tests will be provided, and the benefits and limitations of using urine tests for disease diagnosis will be discussed.
Adi S.
The table shows the data for purification of a hypothetical protein. Determine the missing values for the specific activity, yield, and purification level for the three types of chromatography: | Step | Total protein (mg) | Total activity (units) | Specific activity (units/mg) | Yield (%) | Purification level | |------------------------|--------------------|-----------------------|------------------------------|-----------|--------------------| | Homogenization | 15100 | 1.35 x 10^5 | 8.94 | 100 | 1 | | Salt fractionation | 4610 | 1.23 x 10^5 | 26.7 | 91.1 | 2.99 | | Ion-exchange chromatography | 1.20 x 10^3 | 1.01 x 10^5 | | | | | Gel-filtration chromatography | 67.5 | 70300 | | | | | Affinity chromatography | 3.85 | 45800 | | | |
Dennis H.
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