Postgraduate Course: Preparative Methods for Structural Biology Laboratory Experience (PGBI11113)
|School||School of Biological Sciences
||College||College of Science and Engineering
|Credit level (Normal year taken)||SCQF Level 11 (Postgraduate)
||Availability||Not available to visiting students
|Summary||Students will get a thorough practical training in the chromatographic and processing methods involved in purifying a recombinantly expressed protein to homogeneity. Using LDH-A as the model system, the course will apply a range of techniques to extract and then purify the material to homogeneity. Students will be exposed to several methods of extraction and experience at least three different types of chromatographic methodologies: IMAC, IEX and GF. They will utilise modern LC equipment - AKTAs - to perform the purification and learn how to identify applicable methods for assessing activity and purify and biophysical coherency of their sample throughout the purification process.
Week 1 Introduction to model system to be used (LDH-A), introduction to method design/implementation techniques to be employed through weeks 2 - 10 and summary of assessment.
Week 2 Practical session on protein concentration determination and SDS-PAGE
Week 3 Practical session on LC system operation. Preparation of buffers/reagents for LC work.
Week 4 Lysis/extraction, clarification and analysis of primary input material
Week 5 IMAC/IEX runs - depending on construct system used, PAGE and activity analysis.
Week 6 Sample pooling rationales.
Week 7 GF - runs-1
Week 8 GF - runs-2
Week 9 Final sample pooling rationales, PAGE and activity analysis.
Week 10 Analytical GF - Rh, apparent MR determination and final purity determination.
Course Delivery Information
|Academic year 2022/23, Not available to visiting students (SS1)
|Learning and Teaching activities (Further Info)
Lecture Hours 3,
Supervised Practical/Workshop/Studio Hours 17,
Programme Level Learning and Teaching Hours 2,
Directed Learning and Independent Learning Hours
|Assessment (Further Info)
|Additional Information (Assessment)
||Practical skills and lab-books will be assessed on a weekly basis - 20%
A report of all experiments and results, in the form of a research paper - 80%
||Written and verbal feedback will be provided on practical skills and lab-books during practical sessions throughout the course. Written feedback will be provided on the written report at the end of the course.
|No Exam Information
On completion of this course, the student will be able to:
- Apply a range of chromatographic and extraction techniques to purify and assess proteins.
- Analyse and assess the applicable methods and practical logistics of their sample/methodology in context of the changing experimental conditions and available equipment, compared to the ideal method
- Demonstrate ability to design and implement a mult-step purification protocol and suggest applicable biophysical methods for assessment of sample purity and activity.
- Demonstrate the ability to operate advanced instrumentation and develop dynamic strategies for alteration/adaptation of standard methods based on the behaviour of real samples.
|Graduate Attributes and Skills
||Enhanced practical skills.
|Keywords||Structural Biology,Drug Discovery
|Course organiser||Dr Matthew Nowicki
|Course secretary||Mrs Claire Black
Tel: (0131 6)50 8637