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Application of dye-ligands affinity adsorbent in capturing of rabbit immunoglobulin G


Wongchuphan, Rattana and Tey, Beng Ti and Tan, Wen Siang and Taip, Farah Saleena and Mustapa Kamal, Siti Mazlina and Ling, Tau Chuan (2009) Application of dye-ligands affinity adsorbent in capturing of rabbit immunoglobulin G. Biochemical Engineering Journal, 45 (3). pp. 232-238. ISSN 1369-703X; ESSN: 1873-295X


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.

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Additional Metadata

Item Type: Article
Divisions: Faculty of Biotechnology and Biomolecular Sciences
Faculty of Engineering
Institute of Bioscience
DOI Number: https://doi.org/10.1016/j.bej.2009.04.003
Publisher: Elsevier
Keywords: Affinity; Bioseparation; Dye-ligand; Expanded bed chromatography; Immunoglobulin G purification; Adsorption
Depositing User: Nabilah Mustapa
Date Deposited: 07 May 2015 04:41
Last Modified: 20 Oct 2015 02:07
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1016/j.bej.2009.04.003
URI: http://psasir.upm.edu.my/id/eprint/13225
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