TY - JOUR
T1 - Application of dye-ligands affinity adsorbent in capturing of rabbit immunoglobulin G
AU - Wongchuphan, Rattana
AU - Tey, Beng Ti
AU - Tan, Wen Siang
AU - Taip, Farah Saleena
AU - Kamal, Siti Mazlina Mustapa
AU - Ling, Tau Chuan
N1 - Funding Information:
This study was supported by Grants 07-10-07-406FR and 02-01-04-SF0808. R. Wongchuphan is supported by Suratthani Rajabhat University, Thailand.
Copyright:
Copyright 2009 Elsevier B.V., All rights reserved.
PY - 2009/8/15
Y1 - 2009/8/15
N2 - The applicability of dye-ligands attached to an expanded bed chromatography quartz base matrix (Streamline™) for the affinity bioseparation of rabbit immunoglobulin G (IgG) was investigated. Reactive Green 5 (RG-5) immobilized onto adsorbent was selected for capturing of rabbit-IgG due to its higher binding capacity compared to other dye-ligands possessing similar ligand density. Adsorption parameters such as pH, temperature, ionic strength and initial rabbit-IgG concentration were optimized for the adsorption of rabbit-IgG on the RG-5-immobilized adsorbent. The highest rabbit-IgG adsorption was recorded in pH 7.0, while the maximum binding capacity for BSA was achieved at pH 4.0. The adsorption of rabbit-IgG on RG-5-immobilized adsorbent was declined as the increase of ionic strength. There is no significant influence of temperature against adsorption efficiency of RG-5-immobilized adsorbent for rabbit-IgG. The adsorption phenomenon of rabbit-IgG on RG-5-immobilized adsorbent appeared to follow the Langmuir-Freundlich adsorption isotherm model. The theoretically maximum binding capacity (qm) of RG-5-immobilized adsorbent estimated from this isotherm was 49.3 mg ml-1, which is very close to that obtained experimentally (49.0 mg ml-1). About 50% of bound BSA on RG-5-immobilized adsorbent in binary adsorption system was removed with washing buffer containing 1 M NaCl.
AB - The applicability of dye-ligands attached to an expanded bed chromatography quartz base matrix (Streamline™) for the affinity bioseparation of rabbit immunoglobulin G (IgG) was investigated. Reactive Green 5 (RG-5) immobilized onto adsorbent was selected for capturing of rabbit-IgG due to its higher binding capacity compared to other dye-ligands possessing similar ligand density. Adsorption parameters such as pH, temperature, ionic strength and initial rabbit-IgG concentration were optimized for the adsorption of rabbit-IgG on the RG-5-immobilized adsorbent. The highest rabbit-IgG adsorption was recorded in pH 7.0, while the maximum binding capacity for BSA was achieved at pH 4.0. The adsorption of rabbit-IgG on RG-5-immobilized adsorbent was declined as the increase of ionic strength. There is no significant influence of temperature against adsorption efficiency of RG-5-immobilized adsorbent for rabbit-IgG. The adsorption phenomenon of rabbit-IgG on RG-5-immobilized adsorbent appeared to follow the Langmuir-Freundlich adsorption isotherm model. The theoretically maximum binding capacity (qm) of RG-5-immobilized adsorbent estimated from this isotherm was 49.3 mg ml-1, which is very close to that obtained experimentally (49.0 mg ml-1). About 50% of bound BSA on RG-5-immobilized adsorbent in binary adsorption system was removed with washing buffer containing 1 M NaCl.
KW - Adsorption
KW - Affinity
KW - Bioseparation
KW - Dye-ligand
KW - Expanded bed chromatography
KW - Immunoglobulin G purification
UR - http://www.scopus.com/inward/record.url?scp=67349182598&partnerID=8YFLogxK
U2 - 10.1016/j.bej.2009.04.003
DO - 10.1016/j.bej.2009.04.003
M3 - Article
AN - SCOPUS:67349182598
VL - 45
SP - 232
EP - 238
JO - Biochemical Engineering Journal
JF - Biochemical Engineering Journal
SN - 1369-703X
IS - 3
ER -