Column-free Purification Method for Recombinant Proteins Using a Self-Cleaving Aggregating tag

Bibliographic Details
Main Author:	Miozzi, Jackelyn
Language:	English
Published:	The Ohio State University / OhioLINK 2018
Subjects:	Chemical Engineering
Online Access:	http://rave.ohiolink.edu/etdc/view?acc_num=osu15229401048581

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spelling	ndltd-OhioLink-oai-etd.ohiolink.edu-osu152294010485812021-08-03T07:05:52Z Column-free Purification Method for Recombinant Proteins Using a Self-Cleaving Aggregating tag Miozzi, Jackelyn Chemical Engineering Conventional column-based chromatography processes for the purification of recombinant proteins result in high production costs and slow volumetric throughput at both laboratory and large scale. Non-chromatographic purifications based on selective aggregating tags have the potential to reduce costs with acceptable protein yields. A significant drawback, however, is that current proteolytic approaches for tag removal after the purification are expensive and non-scalable. To address this problem, we have developed a non-chromatographic purification strategy that uses the ¿-roll tag (BRT17) in combination with an engineered split intein for tag removal. The use of the split intein eliminates premature cleavage during expression, which occurs when using a full-length intein such as ¿I-CM, and provides controlled cleavage under mild conditions after purification. The full-length intein aggregating tag was used to effectively purify ¿-lactamase (¿-lac) and resulted in low protein yield due to pre-cleavage. The split intein aggregating tag was used to efficiently purify ¿-lactamase (¿-lac), super-folder green fluorescent protein (sfGFP), streptokinase (SK) and maltose binding protein (MBP), resulting in increased yields compared to previous BRT17-based purifications. 2018-08-14 English text The Ohio State University / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=osu15229401048581 http://rave.ohiolink.edu/etdc/view?acc_num=osu15229401048581 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.
collection	NDLTD
language	English
sources	NDLTD
topic	Chemical Engineering
spellingShingle	Chemical Engineering Miozzi, Jackelyn Column-free Purification Method for Recombinant Proteins Using a Self-Cleaving Aggregating tag
author	Miozzi, Jackelyn
author_facet	Miozzi, Jackelyn
author_sort	Miozzi, Jackelyn
title	Column-free Purification Method for Recombinant Proteins Using a Self-Cleaving Aggregating tag
title_short	Column-free Purification Method for Recombinant Proteins Using a Self-Cleaving Aggregating tag
title_full	Column-free Purification Method for Recombinant Proteins Using a Self-Cleaving Aggregating tag
title_fullStr	Column-free Purification Method for Recombinant Proteins Using a Self-Cleaving Aggregating tag
title_full_unstemmed	Column-free Purification Method for Recombinant Proteins Using a Self-Cleaving Aggregating tag
title_sort	column-free purification method for recombinant proteins using a self-cleaving aggregating tag
publisher	The Ohio State University / OhioLINK
publishDate	2018
url	http://rave.ohiolink.edu/etdc/view?acc_num=osu15229401048581
work_keys_str_mv	AT miozzijackelyn columnfreepurificationmethodforrecombinantproteinsusingaselfcleavingaggregatingtag
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Column-free Purification Method for Recombinant Proteins Using a Self-Cleaving Aggregating tag

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