Purification of aquaporin from spinach and investigation of its efficiency

In this study, it was aimed to purify and determine the efficiency of aquaporin water channel proteins, which play a role in the rapid and active passage of water molecules across biological membranes in many plants and also many other organisms in nature. For the purification of aquaporin, an affinity-based separation column was designed to purify the aquaporin in a single step. For this purpose, first anti-aquaporin antibody nanoparticles were synthesized according to the ANADOLUCA (AmiNoAcid (monomer) Decorated and Light Underpining Conjugation Approach) method and characterized. In nano anti-aquaporin antibody particle synthesis, anti-aquaporin antibody was decorated with photosensitive ruthenium-based amino acid monomer bis(2-2ʹ-bipyridyl)(MATyr)2-ruthenium (II), MATyr-Ru-MATyr, according to ANADOLUCA method and synthesized as nanoparticle by microemulsion polymerization. Then, the synthesized nano anti-aquaporin antibody particles were attached to 2-hydroxyethyl methacrylate (HEMA) cryogel to obtain a specific chromatographic separation column for aquaporin. Aquaporin protein from spinach extract was purified in one step by using the synthesized cryogel column in Fast Protein Liquid Chromatography (FPLC). The molecular weight and the purity of the purified aquaporin, the structural changes during separation were characterized. Thus, in this study, aquaporin was purified in a single step with high purity, with a newly designed reusable column containing specific interaction for purification of aquaporin. Keywords: Aquaporin, ANADOLUCA, Cryogel column, FPLC.