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Biochemistry Laboratory (CHEM 2620)
Total Credits:
2
Lab Credits:
2
Description:
This laboratory course will use an applications-based approach to biochemical techniques. You will apply the concepts learned in the biochemistry lecture course to gain experience in protein purification and characterization, modern molecular biology techniques, enzyme kinetics and immunological techniques. You will extensively utilize biochemical techniques and instrumentation widely used in academic and industry laboratories. You will apply the regulatory affairs standards and document your work as per these standards.
Topical Outline:
1. Preparations of solutions, buffer, and factors that affect protein function and stability (pH, salts, cofactors, detergents, solubilizing agents, reducing agents, chelating agents)
2. NIH RAC guidelines of recombinant DNA processes, workflow of Upstream and Downstream processes in biologics/biopharma, FDA regulations of biochemical products downstream processes
3. Bioinformatics and molecular modeling activities on protein structures, amino acids and peptides; thin layer chromatography of amino acids to assess structure-polarity relationship
4. 9-week research type of project on Recombinant DNA technologies (PCR, restriction digestion, subcloning into appropriate host), expression and purification of protein (ion-exchange, IMAC, size-exclusion and HPLC, and other bioseparations methods)
5. Protein concentration estimations and protein characterization: Bradford, Biurete and BCA assays, specific activity, yield determinations, SDS-PAGE, Western blotting, ELISA, dot Blot, Enzyme kinetics and competitive and non-competitive inhibitions
6. Scientific literature review on the project and writing of scientific report on the project, experimental findings, results, discussions and conclusions, substantiating with the scientific literature references
Learning Outcomes:
1. Demonstrate understanding of the importance of regulations and standards of biochemical processes in biopharma and other industrial applications
2. Successfully design a project and take through all the steps to meet the goal of a biologically active end-product, involving multiple experimental aspects, reagent preparations, documentation, critical thinking of multiple dimensions
3. Carry out laboratory techniques in recombinant protein expression, isolation, and purification using various low and high stringency bioseparation methods including chromatographic techniques
4. Conduct spectrophotometric methods for quantitation of proteins and enzyme kinetics experiments, electrophoretic and immunological methods of protein characterization and be able to explain the underlying scientific concepts
5. Analyze protein sequences using Bioinformatics in understanding protein sequence and structure
6. Maintain record-keeping of experimental work and observations, analyze large sets of data and communicate the outcomes of the project scientific writing format at the level of upper division level writing in undergraduate research
Prerequisites:
CHEM 2224 or
CHEM 2320 or
CHEM 2420 and Have Successfully Completed
CHEM 2610 or register for it concurrently
Co-requisites:
CHEM 2610