1st Assignment (30 Points)
Covers Material Presented in Units 1-3
Section I. (20 Points Total)
The following Topics 1-3 (of 4) cover the cell isolation protocols from Units 1 & 2: Assigned Appendix readings; Isolation of human monocytes by double gradient centrifugation; the Rosette Sep isolation method; and protocols using MACs technology. Your answers should explain the main techniques in your own words. For example: In this protocol the T cells were isolated by incubating them with a CD3-specific antibody that was conjugated with biotin. Following incubation, the excess antibody was removed by washing with buffer and centrifugation. Common protocol details, such as speeds and duration of centrifugation, are not required (e.g., Spin the cells for 30 minutes at 3,000 RPM).
Topic 1. (10 points) Monocyte Isolation
Topic 2. (5points) RosetteSep isolation method (Be sure to watch the video)
In the RosetteSep isolation method, RBCs were not removed before isolation of the cell type of interest.
Topic 3. (5 points) MACS Technology:
Section III. (10 Points) Antibodies for Research and Diagnostic Tools
The following questions are based on Magic Bullets and Monoclonals, FASEB Break Throughs in Bioscience; Appendix I, A-5A12;
Topic 1. (3 points)
Contrast and compare the work of Koch and his postulates and the work of Behring that identified the existence of antibodies. How was Behrings work similar and dissimilar to the work of Robert Koch that resulted in Kochs development of his postulates?
Topic 2: (2 point)
Why was it difficult to isolate the antibodies specific for a disease from the blood of an inoculated animal?
Topic 3. (5 points)
Today, monoclonal antibodies are being successfully used to treat certain cancers (e.g., Rituximab). However, Nadlers and Stashenkos first attempt to treat B cell lymphoma with a monoclonal antibody was a failure.
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