TY - JOUR
T1 - Purification of rabbit polyclonal immunoglobulin G using anion exchangers
AU - Wongchuphan, Rattana
AU - Tey, Beng Ti
AU - Tan, Wen Siang
AU - Subramanian, Senthil Kumar
AU - Taip, Farah Saleena
AU - Ling, Tau Chuan
N1 - Funding Information:
The authors acknowledge Suratthani Rajabhat University, Thailand and Grants 02-01-04-SF0808 and RUGS 91937 for financial support. The authors thank Suet Lin Chia and Ayele Taddese for technical support in the immunization work and Khai Wooi Lee for the purification of HBcAg.
Copyright:
Copyright 2011 Elsevier B.V., All rights reserved.
PY - 2011/1
Y1 - 2011/1
N2 - Negative chromatography antibody purification (N-CAP) using the weak anion exchanger STREAMLINE™ DEAE to extract impurities while retaining the target antibody is proposed as an effective method for the recovery of antibody from rabbit serum. The effects of pH and initial protein concentration on the removal of albumin were investigated. The optimal pH and initial protein concentration for the efficient removal of albumin from rabbit serum were pH 8.0 and 0.5 mg/ml, respectively. Under optimal binding conditions, DEAE successfully removed more than 90% of the albumin from rabbit serum with less than 20% IgG loss. This process offered good polyclonal IgG yield of 80% with a purity of 83% and a purification factor of 5.5. The use of a strong anion exchanger like STREAMLINE™ Q XL for albumin removal was also explored. Under similar optimized conditions, albumin removal by Q XL was as high as 90%. However, IgG recovery and purity were reduced to about 70% and 62%, respectively. Thus, N-CAP using the anion exchanger DEAE removes albumin from rabbit serum and thereby offers an efficient means of purifying polyclonal antibodies.
AB - Negative chromatography antibody purification (N-CAP) using the weak anion exchanger STREAMLINE™ DEAE to extract impurities while retaining the target antibody is proposed as an effective method for the recovery of antibody from rabbit serum. The effects of pH and initial protein concentration on the removal of albumin were investigated. The optimal pH and initial protein concentration for the efficient removal of albumin from rabbit serum were pH 8.0 and 0.5 mg/ml, respectively. Under optimal binding conditions, DEAE successfully removed more than 90% of the albumin from rabbit serum with less than 20% IgG loss. This process offered good polyclonal IgG yield of 80% with a purity of 83% and a purification factor of 5.5. The use of a strong anion exchanger like STREAMLINE™ Q XL for albumin removal was also explored. Under similar optimized conditions, albumin removal by Q XL was as high as 90%. However, IgG recovery and purity were reduced to about 70% and 62%, respectively. Thus, N-CAP using the anion exchanger DEAE removes albumin from rabbit serum and thereby offers an efficient means of purifying polyclonal antibodies.
KW - Albumin removal
KW - Anion exchange adsorbents
KW - Negative chromatography antibody purification
KW - Polyclonal IgG
KW - Rabbit serum
KW - STREAMLINE™ DEAE
UR - http://www.scopus.com/inward/record.url?scp=78650229989&partnerID=8YFLogxK
U2 - 10.1016/j.procbio.2010.07.023
DO - 10.1016/j.procbio.2010.07.023
M3 - Article
AN - SCOPUS:78650229989
SN - 1359-5113
VL - 46
SP - 101
EP - 107
JO - Process Biochemistry
JF - Process Biochemistry
IS - 1
ER -