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Process Development for the Producti...

Corbin, Jasmine Marie.

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Process Development for the Production of Recombinant Butyrylcholinesterase in Metabolically-Regulated, Transgenic Rice Cell Suspension Cultures.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Process Development for the Production of Recombinant Butyrylcholinesterase in Metabolically-Regulated, Transgenic Rice Cell Suspension Cultures./
Author:	Corbin, Jasmine Marie.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 2018,
Description:	275 p.
Notes:	Source: Dissertations Abstracts International, Volume: 80-05, Section: B.
Contained By:	Dissertations Abstracts International80-05B.
Subject:	Pharmacology. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=10928193
ISBN:	9780438628526

Process Development for the Production of Recombinant Butyrylcholinesterase in Metabolically-Regulated, Transgenic Rice Cell Suspension Cultures.
Corbin, Jasmine Marie.

Process Development for the Production of Recombinant Butyrylcholinesterase in Metabolically-Regulated, Transgenic Rice Cell Suspension Cultures. - Ann Arbor : ProQuest Dissertations & Theses, 2018 - 275 p.

Source: Dissertations Abstracts International, Volume: 80-05, Section: B.

Thesis (Ph.D.)--University of California, Davis, 2018.

This item must not be sold to any third party vendors.

The human enzyme butyrylcholinesterase (BChE) has been extensively studied as a therapeutic and prophylactic treatment against organophosphate poisoning and cocaine toxicity, but its use has been limited by the prohibitive costs of production and purification from human plasma (estimated US$20,000 per dose). To address the need for this life-saving drug, we have produced a recombinant version of BChE in a metabolically-regulated transgenic rice cell suspension culture (rrBChE). The culture, described in Chapter 3, is operated using a semicontinuous strategy that allows for independent optimization of growth and production phases, reduces the need for frequent operation of long seed trains, and minimizes turn-around time, CIP and SIP operations, chemicals, and energy. Our data demonstrate that rrBChE can be produced at 10 mg/L and higher through multiple cycles of growth and BChE expression at process scales from 50 mL up to 10 L. To complement this upstream process, we have also developed a downstream processing (Chapter 4) scheme that can recover 42% of active, >95% pure therapeutic-quality rrBChE using widely available unit operations including tangential flow filtration (TFF), diethylaminoethanol (DEAE) anion-exchange chromatography, and affinity chromatography. Use of such operations removes the need for custom equipment, specialized supplies, or significant reconfiguration of existing facilities developed for other cell-based manufacturing scale processes used today. Once pure, we characterized rrBChE's amino acid sequence, enzymatic activity, glycosylation profile, and in vitro inhibition by various organophosphates to confirm its structural and functional comparability to human plasma BChE (Chapter 5). Finally, to assess the impact of these potential cost-saving measures at the manufacturing scale, we scaled up the process in silico using SuperPro Designer (Intelligen, Inc.) and used it to perform a technoeconomic analysis (Chapter 6). Using data obtained from laboratory-scale process optimization experiments, we selected equipment and operational conditions that would enable this specific process to reach realistic production goals for rrBChE. Based on this simulation, this novel process is expected to reduce the cost of each dose nearly 100-fold compared with purification from human plasma BChE, significantly increasing the accessibility of this important drug.

ISBN: 9780438628526Subjects--Topical Terms:

634543
Pharmacology.

Process Development for the Production of Recombinant Butyrylcholinesterase in Metabolically-Regulated, Transgenic Rice Cell Suspension Cultures.
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500 $a Source: Dissertations Abstracts International, Volume: 80-05, Section: B.
500 $a Publisher info.: Dissertation/Thesis.
500 $a Advisor: McDonald, Karen A.
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506 $a This item must not be sold to any third party vendors.
520 $a The human enzyme butyrylcholinesterase (BChE) has been extensively studied as a therapeutic and prophylactic treatment against organophosphate poisoning and cocaine toxicity, but its use has been limited by the prohibitive costs of production and purification from human plasma (estimated US$20,000 per dose). To address the need for this life-saving drug, we have produced a recombinant version of BChE in a metabolically-regulated transgenic rice cell suspension culture (rrBChE). The culture, described in Chapter 3, is operated using a semicontinuous strategy that allows for independent optimization of growth and production phases, reduces the need for frequent operation of long seed trains, and minimizes turn-around time, CIP and SIP operations, chemicals, and energy. Our data demonstrate that rrBChE can be produced at 10 mg/L and higher through multiple cycles of growth and BChE expression at process scales from 50 mL up to 10 L. To complement this upstream process, we have also developed a downstream processing (Chapter 4) scheme that can recover 42% of active, >95% pure therapeutic-quality rrBChE using widely available unit operations including tangential flow filtration (TFF), diethylaminoethanol (DEAE) anion-exchange chromatography, and affinity chromatography. Use of such operations removes the need for custom equipment, specialized supplies, or significant reconfiguration of existing facilities developed for other cell-based manufacturing scale processes used today. Once pure, we characterized rrBChE's amino acid sequence, enzymatic activity, glycosylation profile, and in vitro inhibition by various organophosphates to confirm its structural and functional comparability to human plasma BChE (Chapter 5). Finally, to assess the impact of these potential cost-saving measures at the manufacturing scale, we scaled up the process in silico using SuperPro Designer (Intelligen, Inc.) and used it to perform a technoeconomic analysis (Chapter 6). Using data obtained from laboratory-scale process optimization experiments, we selected equipment and operational conditions that would enable this specific process to reach realistic production goals for rrBChE. Based on this simulation, this novel process is expected to reduce the cost of each dose nearly 100-fold compared with purification from human plasma BChE, significantly increasing the accessibility of this important drug.
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