CHEM 720/7200 - Advanced Bioanalytical Chemistry
Semester: Spring 2019Professor: S. Mikkelsen | Discipline: Analytical | Campus: Waterloo
Introduction to enzymes: classification, nomenclature, specific activity. Review of enzyme kinetics (Michaelis-Menten and pseudo-Michaelis Menten kinetics, conditions for the assay of enzymes, substrates, activators and inhibitors). Enzyme assays, including high-throughput screening (HTS) methods. Immobilization of enzymes: methods including adsorption, entrapment, crosslinking, covalent immobilization and encapsulation; effect of immobilization on kinetics, pH dependence and stability. Immobilized enzyme reactor (IMER) configurations and applications, including lab-on-a-chip devices. Introduction to antibodies: structure, types of natural immunoglobulins, monoclonal and engineered antibodies, characterization of antibody-antigen binding reactions. Immunoassays based on secondary Ab:Ag reactions (including agglutination tests, precipitin tests, single radial immunodiffusion). Quantitative immunoassays based on labels: classification as heterogeneous or homogeneous, label types including radiolabels, fluorophores, enzymes, luminescent and others. Nucleic acid assays and arrays based on immunoassay principles. Biosensors: integration of recognition agents with measurement devices; characterization methods; examples of devices for in vitro and in vivo measurements. Separation methods: zone electrophoresis and isoelectric focussing on gels and in capillaries, immunoassays in capillaries; DNA sequencing; 2D electrophoresis; staining and activity-based detection of separated analytes; chromatographic methods for biomacromolecules including gel filtration and affinity methods; lab-on-a-chip devices incorporating separation and detection stages. Validation of bioanalytical methods and devices.
Prerequisites: Undergraduate Introductory Analytical Chemistry, Instrumental Analysis and Introductory Biochemistry or equivalents, or permission of instructor.
Mikkelsen and Cortón, “Bioanalytical Chemistry,” Second Edition, Wiley, New York, 2016. References to primary literature (journal articles) and other books are contained in this text and (these and others) will be made during lectures. This text contains many examples of past examination questions at the end of each chapter. Two other texts may be useful: Holme and Peck, “Analytical Biochemistry,” 2nd Ed., 1993, and Manz, Pamme and Iossifidis, “Bioanalytical Chemistry,” 2004. The first of these is a lower-level introduction to the topic, while the second considers only selected topics, again in an introductory manner, that are relevant mainly to the second half of this course. These three texts will be placed on reserve in the UW Davis library.
Assignment – 30%
Midterm Exam – 30%
Final Exam – 40%
Assignment Due Date: Last day of lectures
Midterm Examination Thurs May 30, 2019, during the scheduled lecture
Final Examination TBA
One written assignment will be due on the last day of lectures. The assignment is not collaborative, but you are encouraged to discuss your topic with any or all of your classmates. The assignment will involve one recently published original research article (from a good scientific journal) that describes a new bioanalytical method such as an immunoassay, biosensor, or electrophoretic approach. Each student will work with a different article, selected with my input (if needed) and approval. Your topic should employ or suggests arrays, lab-on-chip or multiplexing for high throughput, or be designed for field or point-of-care use. Your written report should summarize the new approach and its validation in one or two pages, and then propose further work in this area, with Plan A and Plan B (in case Plan A does not work), in two or three pages. Reports are to be typed, and can be single-spaced, but must have at least a 12 point font size and one inch (2.5 cm) margins. Five of the total thirty points will be awarded for proper grammar, spelling and use of technical terminology.
- Tue: 2:00 pm - 4:30 pm in C2 278/MacN 203
- Thu: 2:00 pm - 4:30 pm in C2 278/MacN 203