Oriented immobilized anti-hIgG via F(c) fragment-imprinted PHEMA cryogel for IgG purification.
Antibodies are used in many applications, especially as diagnostic and therapeutic agents. Among the various techniques used for the purification of antibodies, immunoaffinity chromatography is by far the most common. For this purpose, oriented immobilization of antibodies is an important step for the efficiency of purification step. In this study, F(c) fragment-imprinted poly(hydroxyethyl methacrylate) cryogel (MIP) was prepared for the oriented immobilization of anti-hIgG for IgG purification from human plasma. Non-imprinted poly(hydroxyethyl methacrylate) cryogel (NIP) was also prepared for random immobilization of anti-hIgG to compare the adsorption capacities of oriented (MIP/anti-hIgG) and random (NIP/anti-hIgG) cryogel columns. The amount of immobilized anti-hIgG was 19.8 mg/g for the NIP column and 23.7 mg/g for the MIP column. Although the amount of immobilized anti-hIgG was almost the same for the NIP and MIP columns, IgG adsorption capacity was found to be three times higher than the NIP/anti-hIgG column (29.7 mg/g) for the MIP/anti-hIgG column (86.9 mg/g). Higher IgG adsorption capacity was observed from human plasma (up to 106.4 mg/g) with the MIP/anti-hIgG cryogel column. Adsorbed IgG was eluted using 1.0 M NaCl with a purity of 96.7%. The results obtained here are very encouraging and showed the usability of MIP/anti-hIgG cryogel prepared via imprinting of Fc fragments as an alternative to conventional immunoaffinity techniques for IgG purification.
Department of Chemistry, Hacettepe University, Ankara, Turkey.
Nilay
N
Bereli
Gizem
G
Ertürk
M Aşkin
MA
Tümer
Ridvan
R
Say
Adil
A
Denizli
D000327:Adsorption; D000888:Antibodies, Anti-Idiotypic / Q000737:chemistry* / Q000378:metabolism; D055910:Antibodies, Immobilized / Q000737:chemistry* / Q000378:metabolism; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D059985:Cryogels / Q000737:chemistry; D017743:Equipment Reuse; D006531:HEPES; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry* / Q000378:metabolism; D007074:Immunoglobulin G / Q000097:blood / Q000302:isolation & purification* / Q000378:metabolism; D054802:Molecular Imprinting / Q000379:methods; D009994:Osmolar Concentration; D011102:Polyhydroxyethyl Methacrylate / Q000737:chemistry*
D000888:Antibodies, Anti-Idiotypic; D055910:Antibodies, Immobilized; D059985:Cryogels; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D011102:Polyhydroxyethyl Methacrylate; D006531:HEPES