title: Fast separation of native proteins using sub-2 μm nonporous silica particles in a chromatographic cake.
abstract: A novel method for the fast separation of native proteins was investigated using sub-2 μm nonporous silica packing inside a chromatographic cake having a diameter much larger than its thickness. Various silica-based particles ranging from 630 nm to 1.2 μm were synthesized and chemically modified with polyethylene glycol 600. The packing material was laterally packed into a series of chromatographic cakes containing the same diameter (10mm) and different thicknesses, ranging from 2 to 10 mm, and tested by hydrophobic interaction chromatography. The results showed that the sub-2 μm NPS particles in a small chromatographic cake were found to have a high efficiency at a flow rate of 10 mL/min and a backpressure of <20 MPa. The effect of the thickness of the chromatographic cake on the resolution of the proteins was also investigated and it was found that too short a column length could dramatically decrease the protein resolution; the minimum column length was also qualitatively evaluated. The presented method is expected to be useful for routine analysis of native and/or intact proteins in hospitals and as a tool for the fast screening protein drugs and optimization of experimental laboratory conditions.
authors.forename: Ruijuan
authors.initials: R
authors.lastname: Niu
authors.forename: Yi
authors.initials: Y
authors.lastname: Min
authors.forename: Xindu
authors.initials: X
authors.lastname: Geng
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000295:instrumentation* / Q000379:methods; D057927:Hydrophobic and Hydrophilic Interactions; D016062:Porosity; D011312:Pressure; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D012822:Silicon Dioxide / Q000737:chemistry*; D013997:Time Factors
chemical_list: D011506:Proteins; D012822:Silicon Dioxide