title: Batch immunoextraction method for efficient purification of aromatic cytokinins.
abstract: A range of benzylaminopurines naturally occur in plants and exhibit high biological activity. Others have been synthesized, such as 6-(2-hydroxy-3-methoxybenzylamino)purine riboside (2OH3MeOBAPR), which has shown interesting anti-cancer activity under in vitro conditions. In order to study the biological activity of this interesting compound in more detail, a rapid and highly efficient method for its purification from complex samples (e.g. blood and plant extracts) is needed. Therefore, we prepared monoclonal antibodies against 2OH3MeOBAPR. The antibody had undetectable cross-reactivity with all natural isoprenoid cytokinins, but relatively high cross-reactivity with aromatic cytokinins as well as some synthetic di- and tri-substituted 6-benzylaminopurines and the corresponding ribosides. The antibody also showed strong responses and specificity in enzyme-linked immunoassays (ELISAs). In addition, it was used to prepare, for the first time, an immunoaffinity sorbent with high specificity and capacity for aromatic cytokinins. A batch immunoextraction method was then developed and optimized for the purification of 2OH3MeOBAPR from murine blood samples. The high efficacy and simplicity of this method (in off-line combination with HPLC-MS) for the isolation of target analytes from biological material is demonstrated in this study.
authors.affiliation: Laboratory of Growth Regulators, Palacký University and Institute of Experimental Botany ASCR, Olomouc, Czech Republic.
authors.forename: Eva
authors.initials: E
authors.lastname: Hauserová
authors.forename: Jana
authors.initials: J
authors.lastname: Swaczynová
authors.forename: Karel
authors.initials: K
authors.lastname: Dolezal
authors.forename: René
authors.initials: R
authors.lastname: Lenobel
authors.forename: Igor
authors.initials: I
authors.lastname: Popa
authors.forename: Marián
authors.initials: M
authors.lastname: Hajdúch
authors.forename: David
authors.initials: D
authors.lastname: Vydra
authors.forename: Kvetoslava
authors.initials: K
authors.lastname: Fuksová
authors.forename: Miroslav
authors.initials: M
authors.lastname: Strnad
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods; D003583:Cytokinins / Q000097:blood / Q000302:isolation & purification*; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C
chemical_list: D003583:Cytokinins