Multimodal peptide ligand extracts parvovirus from interface in affinity aqueous two-phase system

被引:3
作者
Joshi, Pratik U. [1 ,2 ]
Kroger, Stephanie M. [3 ]
Zustiak, Silviya P. [3 ]
Heldt, Caryn L. [1 ,2 ,4 ]
机构
[1] Michigan Technol Univ, Dept Chem Engn, Houghton, MI USA
[2] Michigan Technol Univ, Hlth Res Inst, Houghton, MI USA
[3] St Louis Univ, Dept Biomed Engn, St Louis, MO USA
[4] 1400 Townsend Dr, Houghton, MI 49931 USA
基金
美国国家科学基金会;
关键词
biomanufacturing; extraction; vaccine; virus like particle (VLP); LIQUID-LIQUID-EXTRACTION; PORCINE PARVOVIRUS; PROTEIN SEPARATION; VIRUS; CELL; PURIFICATION; PEG; OPTIMIZATION; PARTICLES; BINDING;
D O I
10.1002/btpr.3338
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Aqueous two-phase systems (ATPS) have found various applications in bioseparations and microencapsulation. The primary goal of this technique is to partition target biomolecules in a preferred phase, rich in one of the phase-forming components. However, there is a lack of understanding of biomolecule behavior at the interface between the two phases. Biomolecule partitioning behavior is studied using tie-lines (TL), where each TL is a group of systems at thermodynamic equilibrium. Across a TL, a system can either have a bulk PEG-rich phase with citrate-rich droplets, or the opposite can occur. We found that porcine parvovirus (PPV) was recovered at a higher amount when PEG was the bulk phase and citrate was in droplets and that the salt and PEG concentrations are high. To improve the recovery, A PEG 10 kDa-peptide conjugate was formed using the multimodal WRW ligand. When WRW was present, less PPV was caught at the interface of the two-phase system, and more was recovered in the PEG-rich phase. While WRW did not significantly increase the PPV recovery in the high TL system, which was found earlier to be optimal for PPV recovery, the peptide did greatly enhance recovery at a lower TL. This lower TL has a lower viscosity and overall system PEG and citrate concentration. The results provide both a method to increase virus recovery in a lower viscosity system, as well as provide interesting thoughts into the interfacial phenomenon and how to recover virus in a phase and not at the interface.
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页数:10
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