Journal
BIOCHEMICAL ENGINEERING JOURNAL
Volume 104, Issue -, Pages 27-33Publisher
ELSEVIER
DOI: 10.1016/j.bej.2015.05.011
Keywords
Monoclonal antibodies; Downstream processing; Purification; Chromatography; Heparin Sepharose
Funding
- Fundacao para a Ciencia e Tecnologia (FCT) [PTDC/EQU-EPR/117427/2010]
- FCT [SFRH/BD/96442/2013, IF/00048/2014]
- Fundação para a Ciência e a Tecnologia [SFRH/BD/96442/2013, PTDC/EQU-EPR/117427/2010] Funding Source: FCT
Ask authors/readers for more resources
In this work, an innovative and cost effective alternative to Protein A based on a commercially available HiTrap (TM) Heparin resin was evaluated as a first capture step for the purification of a monoclonal antibody from different CHO cell culture supernatants. Preliminary studies using pure human serum antibodies and fetal bovine serum showed that heparin can be used as a cation exchanger for the capture of antibodies. Different adsorption and elution buffers were screened and the most promising results were obtained using 20 mM phosphate buffer at pH 8.0 as binding buffer and 1 M NaCl in 20 mM phosphate buffer at pH 8.0 as elution buffer. The performance of the capture step was evaluated in terms of yield, purity, and purification factor of the elution pool obtained. The studies revealed that heparin is capable of an impressive performance, with recoveries higher than 96% from a diafiltered serum-free CHO cell supernatant for all tested conditions (pH 7.0-8.0), and surpassing 99% at pH 8.0. This is especially important considering that the separation did not rely on a highly selective bioaffinity step. The purification factor was found to increase with the pH value, with the highest value (representing a 3.1-fold increase) also obtained at pH 8.0. (C) 2015 Elsevier B.V. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available