Bi1_2011_PS8

Bi1_2011_PS8 - Bi1 The Biology and Biophysics of Viruses...

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Bi1: The Biology and Biophysics of Viruses Spring 2011 Problem Set 8: Vaccines Due Tuesday, May 31 at 4:00 P.M. in the Bi 1 closet Name: ______________________________________________________________ Section # : ___________________________________________________________ Mail Code : ___________________________________________________________ TA Names : ___________________________________________________________ Date and Time turned in : _______________________________________________ Number of pages including this one : _____________________________________ AFTER YOU FINISH: How long did it take you to complete this problem set? ____________________________ Go to the Bi1 moodle site at http://courses.caltech.edu/ and take the homework survey There is 1 question. The number of parts to each question is listed at the beginning of each; be sure to answer all the parts! Grade: Problem 1: __________ TOTAL: _________ HOMEWORK INSTRUCTIONS 1) Turn in your homework stapled to this cover page. 2) Use separate sheets of paper for your answers. 3) Write or type your answers neatly. 4) Put your name on each page of your answers. 5) Box your answers, please, so that the grader can find them. Points may be deducted if you don’t follow these instructions!
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Page 2 of 7 Problem 1: Vaccine Development and Testing (100 points 19 parts) Suggested Readings: 1. Lecture 24 and 25 on Vaccines 2. http://www.ncbi.nlm.nih.gov/pubmed/19843557 Vaccination with ALVAC and AIDSVAX to prevent HIV- 1 infection in Thailand.” (main article) PART I. A patient infected with HIV typically produces antibodies against HIV proteins. Most of these antibodies are non-neutralizing (i.e., they do not prevent infection by HIV). However, the presence of antibodies against HIV proteins can be used in the diagnosis of HIV infection. An individual is said to have unde rgone “seroconversion” if antibodies against HIV can be detected in his or her blood. In the United States, two tests are commonly performed to detect anti-HIV antibodies. The first screening test is an ELISA (enzyme-linked immunosorbent assay) to look for antibodies against the HIV p24 protein. A. (3 points) List the steps required to perform an anti-p24 ELISA (100 words or less). B. (3 points) What is an appropriate negative control for an anti-p24 ELISA? Explain briefly. (2-3 sentences) C. (7 points) How can you evaluate the detection limit of this type of ELISA; i.e., how can you determine the lowest concentration of anti-p24 antibodies in the blood that could be detected? Can an ELISA be used to estimate the concentration of anti-p24 antibodies in the blood of a seroconverted individual? ( 3-4 sentences) If the ELISA test is positive, then a second screening test, called a Western blot, is conducted to confirm the positive result. A Western blot is a general procedure that uses antibodies to detect particular protein(s) from a mixture of proteins that has been separated by a procedure called SDS- p oly a crylamide g el e lectrophoresis (SDS-PAGE). SDS refers to a negatively-charged detergent, sodium dodecyl sulfate, which is incubated with the proteins and binds to them before they are subjected to gel electrophoresis on a polyacrylamide gel. SDS binds to most/all proteins resulting in a constant charge to mass ratio, allowing proteins to be separated by electrophoresis based on their molecular weights.
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