title: Design and synthesis of novel hydrophilic spacers for the reduction of nonspecific binding proteins on affinity resins.
abstract: Tubulin and actin often bind nonspecifically to affinity chromatography resins, complicating research toward identifying the cellular targets. Reduction of nonspecific binding proteins is important for success in finding such targets. We herein disclose the design, synthesis, and effectiveness in reduction of nonspecific binding proteins, of novel hydrophilic spacers (2-5), which were introduced between matrices and a ligand. Among them, tartaric acid derivative (5) exhibited the most effective reduction of nonspecific binding proteins, whilst maintaining binding of the target protein. Introduction of 5 on TOYOPEARL reduced tubulin and actin by almost 65% and 90% compared to that without the hydrophilic spacer, respectively, with effective binding to the target protein, FKBP12.
authors.affiliation: Chemistry Department, Reverse Proteomics Research Institute Co., Ltd, 2-6-7 Kazusa-Kamatari, Chiba 292-0818, Japan.
authors.forename: Takaaki
authors.initials: T
authors.lastname: Shiyama
authors.forename: Minoru
authors.initials: M
authors.lastname: Furuya
authors.forename: Akira
authors.initials: A
authors.lastname: Yamazaki
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Terada
authors.forename: Akito
authors.initials: A
authors.lastname: Tanaka
mesh_terms: D000199:Actins / Q000378:metabolism; D000837:Anion Exchange Resins / Q000737:chemistry* / Q000378:metabolism; D002846:Chromatography, Affinity; D011485:Protein Binding; D011506:Proteins / Q000302:isolation & purification*; D012117:Resins, Synthetic / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D013644:Tartrates / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism
chemical_list: D000199:Actins; D000837:Anion Exchange Resins; D011506:Proteins; D012117:Resins, Synthetic; D013644:Tartrates; D014404:Tubulin; C029768:tartaric acid