title: Antibody affinity purification using metallic nickel particles.
abstract: Functionalized magnetic particles are emerging as a reliable and convenient technique in the purification of biomacromolecules (proteins and nucleic acids) and cell separation. In this study, we used novel solid nickel ferromagnetic particles coated with Protein A for the affinity purification of antibody. The study demonstrated that IgG can be purified from undiluted mouse serum in as few as 5 min using Protein A-coated nickel particles. Further, protein crosslinking was shown to stabilize the Protein A on the nickel particle surfaces to minimize Protein A leaching during the affinity purification and elution of IgG. The separation procedure is gentle, scalable, automatable, efficient and economical. By modifying the functional groups of amino acids in the protein coating, crosslinked nickel particles can be used not only for protein affinity purification but for other biological sample preparation and chromatographic applications as well. Methods proposed and tested in this study can be easily modified for small and medium scale antibody purification in lab and pre-clinical research.
authors.affiliation: Key Lab of Colloid and Interface Chemistry, Ministry of Education, Institute of Theoretical Chemistry, School of Chemistry & Chemical Engineering, Shandong University, Jinan 250100, PR China.
authors.forename: Jun
authors.initials: J
authors.lastname: Gao
authors.forename: Zhijun
authors.initials: Z
authors.lastname: Li
authors.forename: Thomas
authors.initials: T
authors.lastname: Russell
authors.forename: Zhiyu
authors.initials: Z
authors.lastname: Li
mesh_terms: D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity; D052203:Ferrosoferric Oxide / Q000737:chemistry*; D005976:Glutaral; D007074:Immunoglobulin G / Q000302:isolation & purification*; D051379:Mice; D008863:Microspheres; D009532:Nickel / Q000737:chemistry*; D011485:Protein Binding; D012710:Serum Albumin, Bovine; D013205:Staphylococcal Protein A
chemical_list: D007074:Immunoglobulin G; D013205:Staphylococcal Protein A; D012710:Serum Albumin, Bovine; D009532:Nickel; D005976:Glutaral; D052203:Ferrosoferric Oxide