C6270 Methods of Protein Separation - practice

Faculty of Science
Spring 2011 - only for the accreditation
Extent and Intensity
0/0/3. 3 credit(s) (fasci plus compl plus > 4). Type of Completion: z (credit).
Teacher(s)
doc. RNDr. Oldřich Janiczek, CSc. (seminar tutor)
Guaranteed by
doc. RNDr. Oldřich Janiczek, CSc.
Department of Biochemistry – Chemistry Section – Faculty of Science
Prerequisites
( 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) and Methods of Proteins separations (C6260).
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
there are 7 fields of study the course is directly associated with, display
Course objectives
The objective of the practice is to get to students the basic knowledges from separation methods in biochemistry or molecular biology, which are used for protein purification. Contents of the first part includes the basic manipulation with biological samples - extraction, clarification, precipitation, ultrafiltration and lyophilisation. Second part is orientied onto chromatography methods. Last part is dedicated in electromigration methods.
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 Chromatography with ESI-MS. Protein tryptic digest analysis. 9.Gel Permeation Chromatography. 10.Affinity Chromatography. 11.Electromigration Metods. Basic Principles. 12.Free and Zone Electrophoresis. 13.Izoelectric 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
Brief lecture follwed by practical laboratory training.
Assessment methods
Class discussion and written protocols.
Language of instruction
Czech
Follow-Up Courses
Further Comments
The course is taught annually.
The course is taught: every week.
Teacher's information
http://orion.chemi.muni.cz/biochem/student.htm
The course is also listed under the following terms Spring 2001, Spring 2002, Spring 2003, Spring 2004, Spring 2005, 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.