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Development of an aptamer-based affinity purification method for vascular endothelial growth factor

Publikation: Beitrag in FachzeitschriftArtikelForschungPeer-Review

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OriginalspracheEnglisch
Seiten (von - bis)16-23
Seitenumfang8
FachzeitschriftBiotechnology Reports
Jahrgang8
PublikationsstatusVeröffentlicht - 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.

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Development of an aptamer-based affinity purification method for vascular endothelial growth factor. / Lönne, Maren; Bolten, Svenja; Lavrentieva, Antonina et al.
in: Biotechnology Reports, Jahrgang 8, 28.08.2015, S. 16-23.

Publikation: Beitrag in FachzeitschriftArtikelForschungPeer-Review

Lönne M, Bolten S, Lavrentieva A, Stahl F, Scheper T, Walter JG. Development of an aptamer-based affinity purification method for vascular endothelial growth factor. Biotechnology Reports. 2015 Aug 28;8:16-23. doi: 10.1016/j.btre.2015.08.006
Lönne, Maren ; Bolten, Svenja ; Lavrentieva, Antonina et al. / Development of an aptamer-based affinity purification method for vascular endothelial growth factor. in: Biotechnology Reports. 2015 ; Jahrgang 8. S. 16-23.
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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.",
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note = "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).",
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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).

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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

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