Analytical Biochemistry
Theory: 2 hours/week | Tutorials: 2 hours/week | ECTS Units: 5
Tutoring in the English language is offered to Erasmus students
Learning Outcomes
The main aim of the course is to provide the students with a comprehensive description of the principles and the range of applications of basic and advanced experimental techniques of biochemistry, cellular and molecular biology and to practice the students on setting up experimental protocols Upon completion of the course the students are expected to:
- Know and be able to describe the principles of basic and advanced techniques of biochemistry, cellular and molecular biology.
- Distinguish, combine and identify the application of these techniques in both basic and applied research.
- Combine knowledge and available information to design and develop experimental protocols with the use of biochemical tools and processes
- Collaborate with fellow students to design experimental protocols
- Evaluate and analyze results of experimental research
Analytical Description of the Course
- Centrifugation in biochemistry: Principles of centrifugation, Organology of centrifugation, Applications of centrifugation
- Methods of isolation and characterization of biologically active cellular organelles
- Chromatographic techniques for the purification and analysis of macromolecules: Principle of chromatography, Paper and thin layer flat chromatography, Column chromatography, Ion exchange chromatography, Exclusion gel chromatography, High performance chromatography, Affinity and immunosorbent chromatography, Membrane chromatography. Applications of chromatographic techniques.
- Protein electrophoresis: Polyacrylamide gel electrophoresis, Native Electrophoresis, Urea electrophoresis, SDS gel electrophoresis, Isoelectric focusing electrophoresis, Two-dimensional electrophoresis, Capillary electrophoresis, Immunoelectrophoresis, Applications
- Spectroscopic analysis of biomolecules: UV-visible spectroscopy, fluorescence spectroscopy, mass spectrometry, Applications
- Immunochemical methods of analysis: Immunoprecipitation, Immunocytochemistry, Immunocytochemistry, ELISA, Immunoprecipitation, Immunosorbent assay
- Expression of recombinant proteins-Applications
- Confocal microscopy-applications.
- Image analysis – Live imaging. Methods for characterization of biomolecules interaction and mobility (FRET, FRAP, iFRAP, FLIP, etc.).
- Flow cytometry-applications
- Basic and advanced methods for the analysis of protein-protein interactions: Immunoprecipitation, Co-immunoprecipitation, pull-down_system for in vitro expression of labelled proteins in the presence of labelled amino acids, Proximity ligation assay_proximity molecule binding assay, Applications of methods for the identification and characterization of protein-protein interactions
Student Performance Evaluation
The performance of the students is evaluated through written exams at the end of the semester (70%) and through presentation of an assignment before the written exams (30%). The assignment is given to the students at the beginning of the semester and it is based in a scientific publication, with a content relevant to the course. The students are expected to analyze the relevant scientific literature and present the content of the publication in a 50 min session followed by 10 minutes of QA.
Suggested Bibliography
The content of the course is not fully covered by any available textbook in the Greek literature and for this reason there are various relevant textbooks that complementary cover the full content of the course:
- Σύγχρονη Πειραματική Βιοχημεία (2018), Rodney Boyer, Εκδόσεις Κωσταράκη
- Πειραματική Βιοχημεία, CLARK J.M., SWITZER R.L., Ίδρυμα Τεχνολογίας & Έρευνας-Πανεπιστημιακές Εκδόσεις Κρήτης
- Εργαστηριακές Ασκήσεις Βιοχημείας, (2018) 1η έκδοση, Εκδότης: Ζήτη Πελαγία & Σια Ι.Κ.Ε.
- Basic Methods for the Biochemical Lab [electronic resource]
- Surface Plasmon Resonance Based Sensors [electronic resource]
- Psarra AM notes on «Συνεστιακή μικροσκοπία και εφαρμογές – Μέθοδοι χαρακτηρισμού αλληλεπίδρασης Βιομορίων
Teaching Material / E-class
Lecturer
