Plant‐Derived Manufacturing of Apolipoprotein AI<sub>Milano</sub>: Purification and Functional Characterization

Nykiforuk, Cory L.; Shen, Yin; Murray, Elizabeth; Boothe, J. G.; Moloney, Maurice M.

doi:10.1002/9781118356807.ch20

Production of Plasma Proteins for Therapeutic Use 2012

DOI: 10.1002/9781118356807.ch20

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Plant‐Derived Manufacturing of Apolipoprotein AI_Milano: Purification and Functional Characterization

Cory L. Nykiforuk

Yin Shen

Elizabeth Murray

et al.

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2016

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Liquid-Liquid Phase Separation of Oil Bodies from Seeds

Nykiforuk¹

2016

Methods in Molecular Biology

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Fundamentally, oil bodies are discrete storage organelles found in oilseeds, comprising a hydrophobic triacylglycerol core surrounded by a half-unit phospholipid membrane and an outer shell of specialized proteins known as oleosins. Oil bodies possess a number of attributes that were exploited by SemBioSys Genetics to isolate highly enriched fractions of oil bodies through liquid-liquid phase separation for a number of commercial applications. The current chapter provides a general guide for the isolation of oil bodies from Arabidopsis and/or safflower seed, from which protocols can be refined for different oilseed sources. For SemBioSys Genetic's recombinant technology, therapeutic proteins were covalently attached to oleosins or fused in-frame with ligands which bound oil bodies, facilitating their recovery to high levels of purity during "upstream processing" of transformed seed. Core to this technology was oil body isolation consisting of simple manipulation including homogenization of seeds to free the oil bodies, followed by the removal of insoluble fractions, and phase separation to recover the oil bodies. During oil body enrichment (an increase in oil body content concomitant with removal of impurities), a number of options and tips are provided to aid researchers in the manipulation and monitoring of these robust organelles.

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Liquid-Liquid Phase Separation of Oil Bodies from Seeds

Nykiforuk¹

2016

Methods in Molecular Biology

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Plant‐Derived Manufacturing of Apolipoprotein AI_Milano: Purification and Functional Characterization

Cited by 1 publication

References 128 publications

Liquid-Liquid Phase Separation of Oil Bodies from Seeds

Liquid-Liquid Phase Separation of Oil Bodies from Seeds

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Plant‐Derived Manufacturing of Apolipoprotein AIMilano: Purification and Functional Characterization

Cited by 1 publication

References 128 publications

Liquid-Liquid Phase Separation of Oil Bodies from Seeds

Liquid-Liquid Phase Separation of Oil Bodies from Seeds

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Plant‐Derived Manufacturing of Apolipoprotein AI_Milano: Purification and Functional Characterization