18 / 32
Notes:
Page 40
Chromatography 2016
September 21-23, 2016
Volume 7, Issue 5(Suppl)
J Chromatogr Sep Tech 2016
ISSN: 2157-7064 JCGST, an open access journal
conferenceseries
.com
September 21-23, 2016 Amsterdam, Netherlands
World Congress on
Chromatography
Implementation of ssDNAaptamers for imidazole-free purification of His
3
-tagged recombinant proteins
Wojciech Strzałka
Jagiellonian University, Poland
T
he dynamic development of genetic engineering has opened new perspectives for the production of recombinant proteins
which are currently offered by many biotech companies. In addition to the medical and industrial applications they are
also extensively used in basic research studies. Recombinant proteins are often produced in heterologous expression systems,
for example in
E. coli
cells. Before proteins find a final application, purification, a key stage of the production process, must
be performed. Therefore affinity chromatography systems were developed for the fast and simple isolation of recombinant
proteins. One of such systems is Immobilized Metal Ion Affinity Chromatography (IMAC), which is commonly used for the
purification of His
6
-tagged recombinant proteins. Although it is a powerful system it is not free of disadvantages. Recently an
alternative solution, which is free of IMAC drawbacks, was developed. It is based on a unique ssDNA sequence, called the H
3
T
aptamer, which was selected for the purification of His
3
-tagged recombinant proteins. The binding of the H3T aptamer to His3-
tag is controlled by sodium ion concentration. Based on this feature H3T aptamer resins can be successfully employed for the
purification of His3-tagged recombinant proteins from
E. coli
total protein extracts using imidazole-free buffers. The purity of
His3-tagged proteins is superior when purified with the help of the H3T aptamer in comparison with IMAC resins.
Biography
Wojciech Strzałka has completed his PhD at the Jagiellonian University. He completed Post-doc at Salento University, Italy and Osaka University, Japan. Currently,
he is a Group Leader at the Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University. He is studying mechanisms of plant DNA replication
and repair, as well as working on the development of new affinity chromatography systems for the purification of recombinant proteins.
wojciech.strzalka@uj.edu.plWojciech Strzałka, J Chromatogr Sep Tech 2016, 7:5(Suppl)
http://dx.doi.org/10.4172/2157-7064.C1.016