Immunoaffinity microcryogels for purification of transferrin

摘要

Immunoaffinity chromatography has a huge interest in the biomedical and biotechnological fields, in particular for one-step isolation, purification and removal of analyte compounds. In this study, uniform-sized microcryogels, a new type of cryogels, were synthesized using 2-hydroxyetyhl methacrylate and epoxy-group-containing monomer, glycidyl methacrylate for purification of a plasma protein, transferrin. Immunoaffinity microcryogels containing anti-Tf antibodies were characterized by Raman spectroscopy, Fourier transform infrared spectroscopy, optical microscopy, scanning electron microscopy, density measurements and swelling tests. Adsorption studies in aqueous media were carried out in order to examine the effects of medium pH, initial concentration of analyte and contact time. It was found that the optimum pH was 6.0 and the maximum adsorption capacity of immunoaffinity microcryogels at this pH value found to be 9.82 mg/g. The K-L constant for Langmuir isotherm was calculated as 2.65 mL/mg. The maximum adsorption capacity obtained from experimental studies is also very close to the calculated Langmuir adsorption capacity (11.27 mg/g). Langmuir adsorption isotherms and pseudo-second-order kinetic models are consistent with the adsorption process, which means that the adsorption is single layered and chemically controlled. The purity of the eluted hsTf from plasma was about 84% with yield about 82%. After the tenth use of the same microcryogels, the maximum hsTf adsorption capacity decreased by about 20%. The results indicated that the immunoaffinity microcryogels having anti-Tf antibody ligands could be a safe and cost-friendly method for purification of transferrin.