Protein Downstream Processing (PDPE)

Dozent: Prof. Dr. Christian Frech

Semester: 2BME-BPD

Lecture PDPE

Contents:

1. Eukaryotic and prokaryotic protein expression systems

2. Inclusion body formation

3. Protein folding

4. Molecular chaperons and synthetic folding aids

5. Models of in vitro protein refolding

6. Effects on refolding e.g. denaturant concentration, temperature, aggregation

7. Separation principles and techniques in biotechnology

8. Initial fractionation of crude broths

9. Two-phase aqueous-polymer systems

10. Chromatographic methods for protein fractionation and purification

Lab Course DPLE

Contents:

1. Purification of Antibodies

• Antibody Capture by Protein A
• Polishing step- anion exchange chromatography
• Polishing step- CEX with a strong cation exchanger
• Polishing step- HIC

2. Refolding of Lysozyme

• Refolding with DTAB/CTAB
• Oxidative Refolding
• RP-HPLC