BIO281 Biomanufacturing Techniques Lab
BIO281
Biomanufacturing Techniques Lab
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Enzyme Handling & Assay
Protein Assay & Bioreactor Operation
Cell Fractionation & Protein Purification
Analytical Techniques for Purity & Identity
Course Overview
This course provides an additional dimension to Biotechnology by introducing
the students to product-oriented application of the molecular biology
methods they have already studied. The principles learned in this course are
relevant to upscaling for a biomanufacturing processes Using the E. coli ATCC
37658 strain with plasmid-borne ß-galactosidase enzyme activity, the class
provides instruction and experience in spectrophotometric enzyme assay,
growing cells using a BioFlo 110 bioreactor, and in obtaining a purified protein
product that is representative of commercial production procedures. The
content includes the following topics:
Protein structure and properties
Guidelines to handling and working with proteins
Enzyme induction and control
Protein determination and enzymatic assay
Large-scale bacterial growth
Preparation and operation of a bioreactor vessel
Cell fractionation and centrifugation
Strategy and methods for protein purification
Principles of size exclusion and ion exchange column chromatography
Protein gel electrophoresis and staining
Western blot and antibody probe techniques
Page 8
Advanced Biotechnology Certificate Instructional Laboratory Manual
Chapter 6: Molecular Techniques
Experimental procedures for the labs for this course are taken from Protein
Purification Manual, J. Mowry and L. Seidman, Madison Area Technical College.
Specific procedures for induction of ß-galactosidase are from “Experiments in
Molecular Genetics”, Jeffrey H. Miller, Cold Spring Harbor Laboratory, 1972.
Operation of the bioreactor is found in the “Guide to Operations for the BioFlo 110
Fermentor”, New Brunswick Scientific Manual M1273-0054. Other procedures are
available in the Coursework section for each unit. Skills to be developed and assessed
can be found in the BIOHUB.
Learning Outcomes:
The intention is for the student to be able to:
1. Modify single living cells in order to produce desired characteristics.
2. Overcome the challenges of performing successful upscaling
techniques:
 Bioreactor preparation and operation
 Cell harvesting and fractionation techniques
 Protein purification methods, including chromatography
 Enzyme handling and enzyme activity assay methods
 Protein assay methods
 Protein gel electrophoresis techniques
 Western blot/enzyme antibody probe techniques
Course Competencies
Actions that are essential to achieve the learning outcomes:
1. Perform large-scale bacterial cell growth for a specific
biomanufacturing process.
2. Validate the principles and importance of sterility in industrial
fermentations
3. Demonstrate aseptic technique and sterility control procedures
4. Complete external measurement of fermentation parameters
5. Demonstrate preparation of seed inoculums
6. Recover protein biomass
7. Calculate the efficiency of a fermentation process
8. Demonstrate preparation of bioreactor media and seed inoculums
9. Operate a plan, execute and validate bioproduct purification
procedures
10. Comply with regulatory and ecological guidelines in the operation and
design of fermentation processes
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