title: Development of an aptamer-based affinity purification method for vascular endothelial growth factor.
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.
authors.affiliation: Institut für Technische Chemie, Leibniz Universität Hannover, Callinstr. 5, D-30167 Hannover, Germany.
authors.forename: Maren
authors.initials: M
authors.lastname: Lönne
authors.affiliation: Institut für Technische Chemie, Leibniz Universität Hannover, Callinstr. 5, D-30167 Hannover, Germany.
authors.forename: Svenja
authors.initials: S
authors.lastname: Bolten
authors.affiliation: Institut für Technische Chemie, Leibniz Universität Hannover, Callinstr. 5, D-30167 Hannover, Germany.
authors.forename: Antonina
authors.initials: A
authors.lastname: Lavrentieva
authors.affiliation: Institut für Technische Chemie, Leibniz Universität Hannover, Callinstr. 5, D-30167 Hannover, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Stahl
authors.affiliation: Institut für Technische Chemie, Leibniz Universität Hannover, Callinstr. 5, D-30167 Hannover, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Scheper
authors.affiliation: Institut für Technische Chemie, Leibniz Universität Hannover, Callinstr. 5, D-30167 Hannover, Germany.
authors.forename: Johanna-Gabriela
authors.initials: JG
authors.lastname: Walter
keywords: Affinity separation; Aptamer immobilization; Protein purification; VEGF