Purification of rabbit polyclonal immunoglobulin G with ammonium sulphate precipitation and mixed-mode chromatography

S H S Mariam, Chien Wei Ooi, Wen Siang Tan, Janna Ong Binti Abdullah, Arbakariya B Ariff, Beng Ti Tey

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    Abstract

    Immunoglobulins G (IgG) against hepatitis B core antigen (HBcAg) was successfully purified using a purification scheme comprising ammonium sulphate precipitation and SepFast? MM AH-1 column chromatography. Ammonium sulphate precipitation performed at 40 saturation was optimum in terms of the recovered polyclonal IgG concentration (7.8 mg/ml) and the removal of albumin (72 ). The yield, purity and purification factor achieved from this simple purification method were 99 , 94 and 7.8, respectively. The IgG recovered from ammonium sulphate precipitation was subjected to SepFast? MM AH-1 column chromatography and the purity of IgG was further increased to 98 , corresponding to a purification factor of 8.1. Protein aggregation was also reduced significantly in the purified IgG sample. Furthermore, the salt content in the purified sample was reduced by 75 and therefore the need of desalting final product was eliminated. Enzyme-linked immunosorbent assay (ELISA) showed that the antigenicity of anti-HBcAg IgG obtained after these purification processes was maintained.
    Original languageEnglish
    Pages (from-to)133 - 138
    Number of pages6
    JournalSeparation and Purification Technology
    Volume144
    Issue numberApril 2015
    DOIs
    Publication statusPublished - 2015

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