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. 2019 Nov 3;24(21):3973. doi: 10.3390/molecules24213973

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Preparation and characterization of recombinant AAVR proteins. (A) AAVR-His and AAVR-SBP-His proteins were expressed in E.coli. BL21 cells under IPTG induction. (lane 1, lysate from cells without IPTG treatments; 2, lysate of His-tagged AAVR (PKD1-5); 3, lysate supernatant from induced BL21; 4, pellet from AAVR-His expressing BL21; 5, lysate from AAVR-SBP-His expressing cells; 6, supernatant from AAVR-SBP-His positive BL21; 7, pellets from AAVR-SBP-His expressed cells. (B) AAVR-His or AAVR-SBP-His protein purified by nickel affinity chromatography and additional gel filtration chromatography. (C) Purified AAVR recombinant protein bound to AAV virion with high affinity. Soluble recombinant AAVR was tested for the binding to AAV1 capsid by BIAcore sensorgrams. The concentrations of the analytes in triplicates are indicated with the calculated association rate constant (Ka), and dissociation rate constant (Kd) and KD values.