C6260 Methods of Protein Separation

Faculty of Science
Autumn 2023
Extent and Intensity
1/0/0. 1 credit(s) (fasci plus compl plus > 4). Recommended Type of Completion: zk (examination). Other types of completion: k (colloquium).
Teacher(s)
prof. RNDr. Zdeněk Glatz, CSc. (lecturer)
Guaranteed by
prof. RNDr. Zdeněk Glatz, CSc.
Department of Biochemistry – Chemistry Section – Faculty of Science
Supplier department: Department of Biochemistry – Chemistry Section – Faculty of Science
Prerequisites
! C6200 Biochemical Methods && ( C3580 Biochemistry && ( C3600 Biochemistry-lab.course || C3620 Biochemistry-lab.course ))
Basic knowledges from general biochemistry in range of the lectures Biochemistry I (C3181) or Biochemistry(C3580).
Course Enrolment Limitations
The course is also offered to the students of the fields other than those the course is directly associated with.
fields of study / plans the course is directly associated with
Course objectives
At the end of the course students should have the basic knowledges from separation methods in biochemistry or molecular biology, which are used for protein purification including the basic manipulation with biological samples - extraction, clarification, precipitation, ultrafiltration and lyophilisation,chromatography methods and electromigration methods.
Learning outcomes
At the end of this course the students should not only theoretically understand but be able practically apply the methods of disintegration of animal, vegetal and microbial tissues, methods of isolation, purification of proteins, nucleic acids and other biopolymers, methods for determination their purity and molecular properties. They should be able to apply these knowledges in their scientific research working on bachelor, master and PhD theses and finally during their following employment as well.
Syllabus
  • 1.Introduction. Basic Preparation. Purification Strategy. 2.Clarification and Extraction. Centrifugation. 3.Phase Separation. Precipitation and Selective Denaturation. Membrane Separation. 4.Ultrafiltration and Lyophilisation. Laboratory Water Preparation. 5.Chromatography. Basic Principles. 6.Adsorption and Partition Chromatography. 7.Ion-exchange Chromatography. Chromatofocusing. 8.Reverse Phase and Ion Pair Chromatography. Hydrophobic Chromatography 9.Gel Permeation Chromatography. 10.Affinity Chromatography. 11.Electromigration Methods. Basic Principles. 12.Free and Zone Electrophoresis. 13.Isoelectric Focusing. 14.Isotachophoresis.
Literature
  • ANZENBACHER, Pavel and Jan KOVÁŘ. Metody chemického výzkumu pro biochemiky. 1. vyd. Praha: Ministerstvo školství ČSR, 1986, 199 s. info
  • Guide to protein purification. Edited by Murray P. Deutscher. San Diego: Academic Press, 1990, xiii, 894. ISBN 0-12-213585-7. info
  • Protein purification methods : a practical approach. Edited by E. L. V. Harris - S. Angal. Oxford: Oxford University Press, 1989, xvi, 317. ISBN 0199630038. info
  • A practical guide to membrane protein purification. Edited by Gebhard von Jagow - Hermann Schägger. San Diego: Academic Press, 1994, 166 s. ISBN 0127255508. info
Teaching methods
Lectures with a demontration of the taught topic by means of educational videos and programs that are available for the students on web page of this course. The students should have the basic knowledge of the basic methods typically used for protein purification at the end of the course.
Assessment methods
The final exam is in the form of written test, which contains questions from the sample extraction, centrifugation, electromigration and chromatographic methods. The time for the test is one hour, the grades are A to F on the basis of each question classification.
Language of instruction
Czech
Follow-Up Courses
Further comments (probably available only in Czech)
The course is taught only once.
The course is taught: every week.
Listed among pre-requisites of other courses
Teacher's information
http://orion.chemi.muni.cz/biochem/student.htm
The course is also listed under the following terms Spring 2008 - for the purpose of the accreditation, Spring 2011 - only for the accreditation, Spring 2000, Spring 2001, Spring 2002, Spring 2003, Spring 2004, Spring 2005, Spring 2006, Spring 2007, Spring 2008, Spring 2009, Spring 2010, Spring 2011, Spring 2012, spring 2012 - acreditation, Spring 2013, Spring 2014, Spring 2015, Spring 2016, Spring 2017, spring 2018, Spring 2019, Spring 2020, Spring 2021, Spring 2022, Spring 2024, Spring 2025.
  • Enrolment Statistics (Autumn 2023, recent)
  • Permalink: https://is.muni.cz/course/sci/autumn2023/C6260