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Affinity precipitation of a monoclonal antibody from an industrial harvest feedstock using an ELP‐Z stimuli responsive biopolymer
Author(s) -
Sheth Rahul D.,
Jin Mi,
Bhut Bharat V.,
Li Zhengjian,
Chen Wilfred,
Cramer Steven M.
Publication year - 2014
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.25230
Subject(s) - monoclonal antibody , chemistry , chromatography , elution , precipitation , biopolymer , affinity chromatography , downstream processing , recombinant dna , yield (engineering) , antibody , biochemistry , polymer , biology , materials science , organic chemistry , enzyme , physics , meteorology , gene , immunology , metallurgy
In this work, a proof of concept elastin‐like polypeptide‐Z domain fusion (ELP‐Z) based affinity precipitation process is developed for monoclonal antibody (mAb) purification from industrial harvest feeds. Greater than 99% mAb recoveries are obtained during the initial binding step of the process for both pure mAb and the mAb harvest feeds. Great than 90% overall mAb yields are also obtained for the subsequent elution step of the process with no measurable mAb aggregation. The process is shown to result in more than 2 logs of host cell protein (HCP) and more than 4 logs of DNA clearance from the harvest feed. While the overall mAb yield and HCP clearance for the affinity precipitation process was comparable to Protein A chromatography the DNA clearance was clearly superior. Performance is maintained for mAb final elution concentrations up to 20 g/L, demonstrating the ability of the process to both concentrate and purify the mAb. Effective ELP‐Z regeneration is also demonstrated using 0.1 M NaOH with no adverse effect on subsequent capture efficiency. Finally, the reusability of the ELP‐Z construct and robustness of the process is demonstrated for up to three purification‐regeneration cycles with minimal product and impurity carryover and high yields and purity. This work demonstrates that the ELP‐Z based precipitation approach can be successfully employed as an affinity capture step for industrial mAbs. Biotechnol. Bioeng. 2014;111: 1595–1603. © 2014 Wiley Periodicals, Inc.