Robust and selective nano cavities for protein separation: an interpenetrating polymer network modified hierarchically protein imprinted hydrogel.
In the present work, we report a novel method for the reinforcement of hierarchically structured molecularly imprinted polymer (MIP) for the separation of human serum albumin (HSA) and immunoglobulin G (IgG) proteins under pressure driven flow conditions. The template proteins (HSA or IgG) were first physically adsorbed at their isoelectric point on the surface of wide pore silica particles. Thereafter, the pore system was filled with a monomer solution and polymerized to form a lightly crosslinked polyacrylamide network covering the protein template. In order to enhance the rigidity of the hydrogels, different type of crosslinkers such as ethylene glycol dimethacrylate (EGDMA), 3,4-dihydroxyphenethylamine (DA) and methylene-bis-acrylamide (MBA), at least partially interpenetrated into the initially made acrylamide base hydrogel leading to formation of an interpenetrating polymer network (IPN). Then the silica matrix was removed to leave highly porous and reinforced MIP. TGA together with FT-IR and TEM analysis supported the interpenetration of the secondary crosslinkers in the initially formed polymer matrix. The compression property of the modified hydrogels as a function of degree of swelling was in the following order: DA>EGDMA>MBA-IPN-modified hydrogel. Batch binding assay verified the capability of the IPN modified MIPs to capture the target proteins. Moreover, solid phase extraction, HPLC and SDS-PAGE revealed that the EGDMA modified MIP could selectively capture and separate the proteins from human serum or fermentation broth.
Institut fur Umweltforschung (INFU), Technical University of Dortmund, 44221 Dortmund, Germany; Department of Chemical Engineering, University of Mohaghegh Ardabili, 179 Ardabil, Iran. Electronic address: nematollahzadeha@uma.ac.ir.
Ali
A
Nematollahzadeh
Institut fur Umweltforschung (INFU), Technical University of Dortmund, 44221 Dortmund, Germany.
Patrick
P
Lindemann
Institut fur Umweltforschung (INFU), Technical University of Dortmund, 44221 Dortmund, Germany.
Wei
W
Sun
Bibitec GmbH, Universitätsstr. 25, 33615 Bielefeld, Germany.
Jörg
J
Stute
Bibitec GmbH, Universitätsstr. 25, 33615 Bielefeld, Germany.
Dirk
D
Lütkemeyer
Institut fur Umweltforschung (INFU), Technical University of Dortmund, 44221 Dortmund, Germany; Department of Biomedical Sciences, Malmö University, SE-205 06 Malmö, Sweden. Electronic address: borje.sellergren@mah.se.
Börje
B
Sellergren
D000327:Adsorption; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D020100:Hydrogels / Q000737:chemistry; D007074:Immunoglobulin G / Q000032:analysis*; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D054802:Molecular Imprinting / Q000379:methods*; D011108:Polymers / Q000737:chemistry*; D016062:Porosity; D012709:Serum Albumin / Q000032:analysis*; D052616:Solid Phase Extraction / Q000379:methods; D017550:Spectroscopy, Fourier Transform Infrared
D020100:Hydrogels; D007074:Immunoglobulin G; D011108:Polymers; D012709:Serum Albumin; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate
Interpenetrating polymer network; Mechanical strength; Molecularly imprinted polymer; Protein separation