Postgraduate Course: Preparative Methods for Structural Biology (PGBI11026)
|School||School of Biological Sciences
||College||College of Science and Engineering
|Credit level (Normal year taken)||SCQF Level 11 (Postgraduate)
||Availability||Available to all students
|Summary||As structure determination of proteins relies heavily on the supply of suitable protein material, the preparative and analytical methods leading up to X-ray diffraction, NMR or Cyro-EM experiments, for example, are of eminent importance. Commonly utilised methods of, expression, purification of proteins will be discussed. The type and configuration of the instrumentation used in modern protein production labs will be discussed as well as the practical considerations for writing methods on such equipment. As a course outcome, the students should be familiar with all common methods and be able to start practical work with recombinant proteins.
- Source of protein ¿ native tissue vs recombinant expressed (bacterial to mammalian considerations). Expression vector choice for protein production, (prokaryote or eukaryote). Optimisation of yield. Extraction methods, considerations for cell/tissue lysis and primary processing of samples pre-chromatographic steps.
- Chromatographic techniques; theory and applications. Development of protocols depending on final downstream application of protein reagents.
- Applications-1; Purification Strategies and Examples detailing the successful use of multi-step purification methodologies.
- Solubilisation and Re-folding Strategies for producing insoluble Recombinant Proteins. Includes and Examples detailing the successful use of multi-step purification methodologies in the refolding process.
- Applications-2; practical considerations for design of a protein purification protocol for a hypothetical protein.
- Introduction to modern chromatography equipment. Considerations for practical operation and method writing.
Entry Requirements (not applicable to Visiting Students)
||Other requirements|| Entry to the course will be for students already holding (or expecting to hold) an Honours degree or equivalent in a biological subject such as biochemistry, molecular biology, biophysics, pharmacology. Students with a background in chemistry, physics, computer science or informatics will also be considered and those with other scientific or medical backgrounds should contact the Course Organiser.
|Additional Costs|| None
Information for Visiting Students
|High Demand Course?
Course Delivery Information
|Academic year 2015/16, Available to all students (SV1)
|Learning and Teaching activities (Further Info)
Lecture Hours 30,
Programme Level Learning and Teaching Hours 2,
Directed Learning and Independent Learning Hours
|Assessment (Further Info)
|Additional Information (Assessment)
||- Prepared seminar on a methods paper (randomly assigned) as pair. 10 - 12 mins presentation; 3 - 5 minutes questions. Week 10. (50%).
- Written report of a chromatography technique, describing basic theory and the general use of the technique. Give at least 2 examples from the literature of how the technique was successfully used as part of a purification protocol. No more than 2,000 words (not incl. figures and tables and references). (25%).
- Write-up of ProteinLab practical session. Write-up to be constructed like the materials and methods section of a basic research methods paper. Includes figures etc. (15%).
- Analyse a flawed method for purifying a hypothetical protein on an automated liquid chromatography unit using the UNICORN software, and suggest where the method could be improved, both in terms of the instrumentation parameters and the protein specific methodology (10%).
|No Exam Information
On completion of this course, the student will be able to:
- ¿ Have knowledge and appreciation of commonly used strategies for protein production for structural biology and analytical biochemical assays.
- ¿ Be able to understand and assimilate data and relating to protein production and purification methodology design and execution.
- ¿ Be able to understand and extract context specific information about the method used and the post-run processing options from a protein purification LC chromatogram.
¿ Protein Purification Techniques: A Practical Approach (Practical Approach Series). 2nd Edition. Simon Roe, Ed.
¿ Protein Purification Applications: A Practical Approach. 2nd Edition. Simon Roe, Ed.
¿ ¿Protein purification: principles, high-resolution methods, and applications. 2nd Edition. Jan-Christer Janson, Lars Rydén.¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿¿
Advised Preparatory Work
All students should endeavour to learn the structures and basic biophysical properties of all the amino acids and nucleotide bases before the course starts.
|Graduate Attributes and Skills
|Course organiser||Dr Martin Wear
|Course secretary||Miss Vicky Mactaggart
Tel: (0131 6)51 7052
© Copyright 2015 The University of Edinburgh - 18 January 2016 4:32 am