Protein technology and proteome analysis

Course Code :2002FBDBIC
Study domain:Biotechnology
Academic year:2017-2018
Semester:1st semester
Contact hours:60
Study load (hours):168
Contract restrictions: Exam contract not possible
Language of instruction:English
Exam period:exam in the 1st semester
Lecturer(s)Sylvia Dewilde
Xaveer Van Ostade

3. Course contents *

The theoretical aspects will be demonstrated using practica. In the first part of the course, the focus will be on the study of proteins:

-     methods for protein extraction

-     chemical modification of proteins: e.g. (i) chemical modification of the amino acids cysteine or lysine by vinylpiridinylation and modification by maleinic acid respectively, (ii) biotinylation of proteins

-     purification of protein mixtures with limited complexity using ion-exchange and reversed phase chromatography. The purity can be checked using SDS-PAGE

-         making a strategy for cleaving proteins

-         separation of limited complex peptide mixtures using reversed phase chromatography

-     N-terminal sequencing of proteins and peptides by Edman degradation

-         Examples of affinity-chromatography


In the following part of the course, proteomics will be discussed. Focus will be on:

-           separation techniques of complex protein mixtures using gelelectrophoreses or chromatography

-           identification techniques based on mass-spectrometry

-           discussion of different strategies for comparing protein-profiles (ICAT, iTRAQ, DIGE, O16/O18 labelling)


These theoretical lectures will be followed by practical exercises: i) labwork (sample handling and gel electrophoreses), ii) demonstrative practical work (2D-microcapilliary chromatography and mass-spectrometry).