title: Phosphorylcholine (PC)-bonded protein A affinity chromatographic medium for high-throughput purification with reduced non-specific protein adsorption.
abstract: A protein A affinity chromatographic medium based on porous silica modified with phosphorylcholine (PC) groups and amino groups (PNSP) was synthesized. The PC groups functioned as suppressors of non-specific protein adsorption. Recombinant protein A was bound to the amino groups on PNSP with a glutaraldehyde used as a spacer (PNSP-PA). The PC groups and amino groups were immobilized on porous-silica particles using two silane coupling reagents, PC-bound silane, and 3-aminopropyltrimethoxysilane. After optimizing various factors in the synthetic process, the resultant protein A medium showed improvements in non-specific protein adsorption, dynamic binding capacity, and chemical stability under basic conditions compared with conventional protein A affinity media.
authors.affiliation: Shiseido Research Center, 2-2-1 Hayabuchi, Tsuzuki, Yokohama 224-8558, Japan. zhanglantong@263.net
authors.forename: Katsuyuki
authors.initials: K
authors.lastname: Maeno
authors.forename: Aya
authors.initials: A
authors.lastname: Hirayama
authors.forename: Kenichi
authors.initials: K
authors.lastname: Sakuma
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Miyazawa
mesh_terms: D000327:Adsorption; D000818:Animals; D002417:Cattle; D002645:Chickens; D002846:Chromatography, Affinity / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D010767:Phosphorylcholine / Q000737:chemistry*; D016062:Porosity; D011485:Protein Binding; D011506:Proteins / Q000302:isolation & purification* / Q000378:metabolism; D012821:Silanes / Q000737:chemistry; D012822:Silicon Dioxide / Q000737:chemistry; D012972:Sodium Hydroxide / Q000737:chemistry; D013205:Staphylococcal Protein A / Q000737:chemistry* / Q000378:metabolism
chemical_list: D011506:Proteins; D012821:Silanes; D013205:Staphylococcal Protein A; D010767:Phosphorylcholine; D012972:Sodium Hydroxide; D012822:Silicon Dioxide