Chromatographic purification of proteins - 26125

Welcome to the DTU Chemistry course Experimental Chromatographic Purification of Recombinant Proteins - 26125. 

You will be working in pairs. Each day you will spend half a day in the lab and half a day studying the theoretical concepts, preparing for the experimental exercises, and evaluating your results. The course schedule is as follows.

Active participation in all exercises is required (in other words attendance every day from 8.00 am to 5 pm is compulsory) in order to attend the final oral exam.

Daily Schedule X-1 team:

Day
 6  8 10  11 12 13 14 15
8-12

Introduction

Note! Day 1 we will start at 9.00 am

Introduction
to Making buffers
and make buffer

LAB

IEX
plasto-
cyanin




LAB

HR-IEX
plasto-
cyanin




LAB

HIC scouting

 

LAB

MBP-
GFP
AC




LAB

Case 1
Making buffers



LAB
Case 1
MBP-TPH
AC w/ on-
column
cleavage


LAB

Packing
of FF
column




LAB

Case 1
SDS-page
of TPH
purification
 


LAB

Case 2
FXD
FF
IEX



LAB            

Case 2
FXD UF




LAB
Case 2
FXD
IEX
30Q



LAB
Case 2
Evaluate
results





Oral
Exam
30 min
per
student

12.15-
12.45
 Lunch meetings with follow up and questions
13-17 RA and
getting to
know the equipment


LAB

IEX concept

Puri.
strategy
concept

HIC
concept

and
Cell lysis
concepts



AC
concept




GF
concept

and
UF
concept

MBP-GFP GF



LAB
Case 1
MBP-TPH GF/UF/Liq N2 storage

LAB

SDS-Page
concept

 

Case 2
Prepare
FXD
purifi-
cation

Understand
and discuss
all concepts

Case 2
FXD
GF

             

LAB

CIP FF
column
peristaltic
pump


LAB         
           

Prepare
exam
presentation




Abbreviations:  
RA: Risk assessment
IEX: Ion-exchange
HR-IEX: High-resolution IEX
HIC: Hydrophobic interaction chromatography
AC:Affinity chromatography
GF: Gel filtration
UF: Ultrafiltration
MBP: maltose binding protein
TPH: Tryptophan hydroxylase
FF: Fast flow
FXD: Ferredoxin
CIP: Cleaning in place
SDS-Page: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis




http://www.kemi.dtu.dk/uddannelse/elearning/kursus26125_chromatographicpurpro-juni18
18 NOVEMBER 2018