TY - JOUR
T1 - Purification of rabbit polyclonal immunoglobulin G with ammonium sulphate precipitation and mixed-mode chromatography
AU - Mariam, S H S
AU - Ooi, Chien Wei
AU - Tan, Wen Siang
AU - Abdullah, Janna Ong Binti
AU - Ariff, Arbakariya B
AU - Tey, Beng Ti
PY - 2015
Y1 - 2015
N2 - 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.
AB - 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.
U2 - 10.1016/j.seppur.2015.02.012
DO - 10.1016/j.seppur.2015.02.012
M3 - Article
VL - 144
SP - 133
EP - 138
JO - Separation and Purification Technology
JF - Separation and Purification Technology
SN - 1383-5866
IS - April 2015
ER -
