Histone-dependent IgG conservation in octanoic acid precipitation and its mechanism
- 403 Downloads
Octanoic acid (OA) precipitation has long been used in protein purification. Recently, we reported a new cell culture clarification method for immunoglobulin G (IgG) purification, employing an advance elimination of chromatin heteroaggregates with a hybrid OA-solid phase system. This treatment reduced DNA more than 3 logs, histone below the detection limit (LOD), and non-histone host cell proteins (nh-HCP) by 90 % while conserving more than 90 % of the IgG monomer. In this study, we further investigated the conservation of IgG monomer and antibody light chain (LC) to the addition of OA/OA-solid phase complex, with or without histone and DNA in different combinations. The results showed that highly basic histone protein was the prime target in OA/OA-solid phase precipitation system for IgG purification, and the selective conservation of IgG monomer in this system was histone dependent. Our findings partially support the idea that OA works by sticking to electropositive hydrophobic domains on proteins, reducing their solubility, and causing them to agglomerate into large particles that precipitate from solution. Our findings also provide a new perspective for IgG purification and emphasize the necessity to re-examine the roles of various host contaminants in IgG purification.
KeywordsOctanoic acid Agglomeration Histone IgG Light chain
This work was financially supported by QIBEBT (Qingdao Institute of Bioenergy and Bioprocess Technology) Start-up Fund (No. Y571061905) and also by the Biomedical Research Council of A*STAR and Exploit Technologies Pte Ltd., of Singapore (No. ETPL/12-R15GAP-0009).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Budavari S, O’Neil MJ, Smith A, Heckelman PE, Kinneary JF (1996) The Merck index: an encyclopedia of chemicals, drugs, and biologicals (12th ed.) Merck & Co. ISBN 0911910123Google Scholar
- Gagnon P, Nian R, Lee J, Tan L, Abdul Latiff SM, Lim CL, Chuah C, Yang YS, Gan HT (2014a) Nonspecific interactions of chromatin with immunoglobulin G and protein A, and their impact on purification performance. J Chromatogr A 1340:68–78. doi: 10.1016/j.chroma.2014.03.010 CrossRefPubMedGoogle Scholar
- Gagnon P, Nian R, Tan L, Cheong J, Yeo V, Yang YS, Gan HT (2014b) Chromatin-mediated depression of fractionation performance on electronegative multimodal chromatography media, its prevention, and ramifications for purification of immunoglobulin G. J Chromatogr A 1374:145–155. doi: 10.1016/j.chroma.2014.11.052 CrossRefPubMedGoogle Scholar
- Gagnon P, Nian R, Yang YS, Yang Q, Lim CL (2015) Non-immunospecific association of immunoglobulin G with chromatin during elution from protein a inflates host contamination, aggregate content, and antibody loss. J Chromatogr A 1408:151–160. doi: 10.1016/j.chroma.2015.07.017 CrossRefPubMedGoogle Scholar
- Herzer S, Bhangale A, Barker G, Chowdhary I, Conover M, O’Mara BW, Tsang L, Wang SY, Krystek SR Jr, Yao Y, Rieble S (2015) Development and scale-up of the recovery and purification of a domain antibody fc fusion protein-comparison of a two and three-step approach. Biotechnol Bioeng 112:1417–1428. doi: 10.1002/bit.25561 CrossRefPubMedGoogle Scholar
- Lide DR (1990) CRC handbook of chemistry and physics (70th Ed.). CRC Press, Boca Raton (FL)Google Scholar
- Open drug and drug target database. (2005) http://www.drugbank.ca/drugs/DB00072
- Zheng J, Wang L, Twarowska B, Laino S, Sparks C, Smith T, Russell R, Wang M (2015) Caprylic acid-induced impurity precipitation from protein a capture column elution pool to enable a two chromatography step process for monoclonal antibody purification. Biotechnol Prog 259:1515–1525. doi: 10.1002/btpr.2154 CrossRefGoogle Scholar