Details
| Original language | English |
|---|---|
| Pages (from-to) | 16-23 |
| Number of pages | 8 |
| Journal | Biotechnology Reports |
| Volume | 8 |
| Publication status | Published - 28 Aug 2015 |
Abstract
Since aptamers bind their targets with high affinity and specificity, they are promising alternative ligands in protein affinity purification. As aptamers are chemically synthesized oligonucleotides, they can be easily produced in large quantities regarding GMP conditions allowing their application in protein production for therapeutic purposes. Several advantages of aptamers compared to antibodies are described in general within this paper. Here, an aptamer directed against the human Vascular Endothelial Growth Factor (VEGF) was used as affinity ligand for establishing a purification platform for VEGF in small scale. The aptamer was covalently immobilized on magnetic beads in a controlled orientation resulting in a functional active affinity matrix. Target binding was optimized by introduction of spacer molecules and variation of aptamer density. Further, salt-induced target elution was demonstrated as well as VEGF purification from a complex protein mixture proving the specificity of protein-aptamer binding.
Keywords
- Affinity separation, Aptamer immobilization, Protein purification, VEGF
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Immunology and Microbiology(all)
- Applied Microbiology and Biotechnology
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In: Biotechnology Reports, Vol. 8, 28.08.2015, p. 16-23.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Development of an aptamer-based affinity purification method for vascular endothelial growth factor
AU - Lönne, Maren
AU - Bolten, Svenja
AU - Lavrentieva, Antonina
AU - Stahl, Frank
AU - Scheper, Thomas
AU - Walter, Johanna Gabriela
N1 - Funding information: We thank Moran Jerabek-Willemsen (NanoTemper Technologies GmbH) for the advice and for the opportunity for the MST measurements. This work is supported by funding from the Deutsche Forschungsgemeinschaft (DFG) for the Cluster of Excellence REBIRTH (From Regenerative Biology to Reconstructive Therapy).
PY - 2015/8/28
Y1 - 2015/8/28
N2 - Since aptamers bind their targets with high affinity and specificity, they are promising alternative ligands in protein affinity purification. As aptamers are chemically synthesized oligonucleotides, they can be easily produced in large quantities regarding GMP conditions allowing their application in protein production for therapeutic purposes. Several advantages of aptamers compared to antibodies are described in general within this paper. Here, an aptamer directed against the human Vascular Endothelial Growth Factor (VEGF) was used as affinity ligand for establishing a purification platform for VEGF in small scale. The aptamer was covalently immobilized on magnetic beads in a controlled orientation resulting in a functional active affinity matrix. Target binding was optimized by introduction of spacer molecules and variation of aptamer density. Further, salt-induced target elution was demonstrated as well as VEGF purification from a complex protein mixture proving the specificity of protein-aptamer binding.
AB - Since aptamers bind their targets with high affinity and specificity, they are promising alternative ligands in protein affinity purification. As aptamers are chemically synthesized oligonucleotides, they can be easily produced in large quantities regarding GMP conditions allowing their application in protein production for therapeutic purposes. Several advantages of aptamers compared to antibodies are described in general within this paper. Here, an aptamer directed against the human Vascular Endothelial Growth Factor (VEGF) was used as affinity ligand for establishing a purification platform for VEGF in small scale. The aptamer was covalently immobilized on magnetic beads in a controlled orientation resulting in a functional active affinity matrix. Target binding was optimized by introduction of spacer molecules and variation of aptamer density. Further, salt-induced target elution was demonstrated as well as VEGF purification from a complex protein mixture proving the specificity of protein-aptamer binding.
KW - Affinity separation
KW - Aptamer immobilization
KW - Protein purification
KW - VEGF
UR - http://www.scopus.com/inward/record.url?scp=84940972298&partnerID=8YFLogxK
U2 - 10.1016/j.btre.2015.08.006
DO - 10.1016/j.btre.2015.08.006
M3 - Article
AN - SCOPUS:84940972298
VL - 8
SP - 16
EP - 23
JO - Biotechnology Reports
JF - Biotechnology Reports
ER -