title: Affinity precipitation of a monoclonal antibody from an industrial harvest feedstock using an ELP-Z stimuli responsive biopolymer.
abstract: 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.
authors.affiliation: Department of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 110 8th Street, Troy, New York, 12180; Biologics Process Development, Bristol-Myers Squibb, East Syracuse, New York.
authors.forename: Rahul D
authors.initials: RD
authors.lastname: Sheth
authors.forename: Mi
authors.initials: M
authors.lastname: Jin
authors.forename: Bharat V
authors.initials: BV
authors.lastname: Bhut
authors.forename: Zhengjian
authors.initials: Z
authors.lastname: Li
authors.forename: Wilfred
authors.initials: W
authors.lastname: Chen
authors.forename: Steven M
authors.initials: SM
authors.lastname: Cramer
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D011232:Chemical Precipitation; D002846:Chromatography, Affinity / Q000379:methods*; D002850:Chromatography, Gel / Q000379:methods; D056148:Chromatography, Reverse-Phase / Q000379:methods; D004549:Elastin / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D013205:Staphylococcal Protein A / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D004549:Elastin
keywords: DNA clearance; HCP clearance; affinity precipitation; elastin-like polypeptides; harvest feedstock; monoclonal antibodies