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KMID : 1094720130180061169
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Biotechnology and Bioprocess Engineering
2013 Volume.18 No. 6 p.1169 ~ p.1175
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New hydrophobic charge-induction resin with 2-mercaptoimidazole as the ligand and its separation characteristics for porcine IgG
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Phottraithip Wimonrat
Lin Dong-Qiang
Shi Fei
Yao Shan-Jing
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Abstract
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New hydrophobic charge-induction chromatography (HCIC) resin coupled with 2-mercaptoimidazole (MI) as the ligand was prepared and used to purify IgG from porcine plasma. The cellulose matrix was activated by divinylsulfone (DVS), and was then coupled with MI as the functional ligand. The reaction conditions were optimized as pH: 11, volume ratio of DVS to matrix: 1.0, reaction time: 4 h. The ligand density reached about 100 μmol/mL gel. The adsorption isotherms of porcine IgG was determined at different pH values, and high saturated adsorption capacities of 78.02 mg IgG/mL gel were found at pH 8. The adsorption of IgG showed a typical pHdependent property of HCIC, and the adsorption capacity decreased significantly in acidic conditions. The prepared resin was used to separate IgG from porcine plasma. After precipitating the fibrinogen by salting-out, the supernatant was loaded onto the column at pH 7 and the elution pH was optimized. The results indicated that acidic elution pH was necessary to recover the IgG. The purity of IgG in the elution fractions was in the range of 81 ∼ 90%, which demonstrated that HCIC with the new ligand showed the excited separation performs and is a potential effective technique to purify IgG from the complex feedstock.
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KEYWORD
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hydrophobic charge-induction chromatography, immunoglobulin G, 2-mercaptoimidazole, porcine plasma, separation
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