FINAL EXAM 2009 - FINAL EXAM BME 339/CHE 339/BIO 335 FALL...

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FINAL EXAM BME 339/CHE 339/BIO 335 FALL 2009 Friday , December 11, 2009 EID _________________________________________ YOUR NAME _________________________________ The exam consist of three parts: 1. Short answer questions (65 points); 2. Multiple choice (questions mainly from the quizzes) 15 points. 3. Problems (20 points). SECTION 1: Short Answer questions 1. Bioreactors and animal cells (8 points) A. Explain using the appropriate questions how you would carry out an experiment to determine the Monod parameters μ max and K s from experiments where you grow cells in shake flasks (4 points ). B. Similarly explain how you can determine the Monod parameters μ max and K s using a chemostat (3 points). C . Why are animal cells grown in incubators or reactors with 5% CO 2 in the air? (1 point).
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Name:_____________ EID:_____________ 2. Oxygen Transfer (7points total) A. What is the critical dissolved oxygen concentration? (1 point). B. How does the Oxygen Transfer Rate (OTR) depend on the interfacial area and the driving force for transfer? Write the equation and explain the meaning of ALL the terms (2 points). C. At steady state OTR=OUR. What are three ways to make sure that the dissolved oxygen concentration remains constant as the biomass increases during the course of the fermentation? (3 points) . D. Mammalian cells do not have cell walls and therefore they are particularly susceptible to damage in fermenters. How can cell damage be prevented when the cells are grown in high cell density bioreactors (1 point).
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Name:_____________ EID:_____________ 3. Bioseparations (5 points) A. Many important proteins are isolated from human serum. The first step is usually salt fractionation (sometimes called fractionational precipitation). Explain in 2-3 lines what this is. ( 3 points) B The next step is isoelectric point precipitation. Describe what this is in one sentence (1 point) B. Immunoaffinity chromatography is then use to further separate serum proteins that are present in low concentrations such as the anti-clotting protein Factor X. Explain in one sentence how this technique works (1 point). 4. Cloning/Expression (14 points total): A. The human SIRT1 protein plays a critical for aging. You know that the gene contains several introns and is about 5 kb in size. Fortunately the SIRT1 protein has been purified from cows in large amounts and you suspect that the cow and human proteins are very similar. Describe how you can clone the coding sequence of the SIRT1 gene (5 points).
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Name:_____________ EID:_____________ B. You now try to express the protein in bacteria. You need to make as much protein as possible and moreover it is of course important that your produce active protein. You suspect translation of the SIRT1 gene might be a problem. What could be the reason(s) for that and how could this problem be fixed (3 points) C. ALSO you have information that the protein might be toxic and as soon it is produced the cell can no longer divide although it can still make protein. What do you need to
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FINAL EXAM 2009 - FINAL EXAM BME 339/CHE 339/BIO 335 FALL...

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