Comparative evaluations of bulk seeded protein crystallization in batch versus continuous slug flow crystallizers

The present work evaluated for the first time the use of continuous slug flow crystallizer (CSFC) in bulk seeded protein crystallization, using lysozyme (LYZ) as the model protein. LYZ crystals pre-prepared in batch crystallizer were used as seeds. The crystallization was performed at supersaturatio...

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Main Authors: Pu, Siyu, Hadinoto, Kunn
Other Authors: School of Chemical and Biomedical Engineering
Format: Article
Language:English
Published: 2022
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Online Access:https://hdl.handle.net/10356/160115
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Institution: Nanyang Technological University
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spelling sg-ntu-dr.10356-1601152022-07-13T01:34:22Z Comparative evaluations of bulk seeded protein crystallization in batch versus continuous slug flow crystallizers Pu, Siyu Hadinoto, Kunn School of Chemical and Biomedical Engineering Engineering::Chemical engineering Continuous Crystallization Protein Crystallization The present work evaluated for the first time the use of continuous slug flow crystallizer (CSFC) in bulk seeded protein crystallization, using lysozyme (LYZ) as the model protein. LYZ crystals pre-prepared in batch crystallizer were used as seeds. The crystallization was performed at supersaturation level below the metastable-zone-limit. The resultant crystal size distribution (CSD), CSD's reproducibility, LYZ's bioactivity, and crystallization efficiency as characterized by the space-time-yield (STY, mg/h·L) were determined. The results showed that the CSFC's performance was governed by the flowrate, where a trade-off existed between crystal quality and crystallization efficiency upon varying the flowrate. At low flowrates, which reduced the shear rate and prolonged the residence time, well-defined large tetragonal crystals were produced attributed to suppressed secondary nucleation and extended crystal growth, but low STY due to difficulty in transporting the seeds/products crystals. Higher flow rates led to higher STY, but increased production of small non-tetragonal crystals that formed agglomerates. Compared to batch crystallizer, the CSFC produced LYZ crystals of similar average size, morphology, and bioactivity, but with roughly 50% lower STY, as the batch's high-shear environment promoted secondary nucleation, hence higher crystallization rate. The CSD's width and reproducibility was nevertheless significantly improved in CSFC. Ministry of Education (MOE) The authors would like to acknowledge the funding from Ministry of Education Singapore under Academic Research Fund Tier 1RG82/20 (2021). 2022-07-13T01:34:22Z 2022-07-13T01:34:22Z 2021 Journal Article Pu, S. & Hadinoto, K. (2021). Comparative evaluations of bulk seeded protein crystallization in batch versus continuous slug flow crystallizers. Chemical Engineering Research and Design, 171, 139-149. https://dx.doi.org/10.1016/j.cherd.2021.05.011 0263-8762 https://hdl.handle.net/10356/160115 10.1016/j.cherd.2021.05.011 2-s2.0-85106467750 171 139 149 en RG82/20 Chemical Engineering Research and Design © 2021 Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic Engineering::Chemical engineering
Continuous Crystallization
Protein Crystallization
spellingShingle Engineering::Chemical engineering
Continuous Crystallization
Protein Crystallization
Pu, Siyu
Hadinoto, Kunn
Comparative evaluations of bulk seeded protein crystallization in batch versus continuous slug flow crystallizers
description The present work evaluated for the first time the use of continuous slug flow crystallizer (CSFC) in bulk seeded protein crystallization, using lysozyme (LYZ) as the model protein. LYZ crystals pre-prepared in batch crystallizer were used as seeds. The crystallization was performed at supersaturation level below the metastable-zone-limit. The resultant crystal size distribution (CSD), CSD's reproducibility, LYZ's bioactivity, and crystallization efficiency as characterized by the space-time-yield (STY, mg/h·L) were determined. The results showed that the CSFC's performance was governed by the flowrate, where a trade-off existed between crystal quality and crystallization efficiency upon varying the flowrate. At low flowrates, which reduced the shear rate and prolonged the residence time, well-defined large tetragonal crystals were produced attributed to suppressed secondary nucleation and extended crystal growth, but low STY due to difficulty in transporting the seeds/products crystals. Higher flow rates led to higher STY, but increased production of small non-tetragonal crystals that formed agglomerates. Compared to batch crystallizer, the CSFC produced LYZ crystals of similar average size, morphology, and bioactivity, but with roughly 50% lower STY, as the batch's high-shear environment promoted secondary nucleation, hence higher crystallization rate. The CSD's width and reproducibility was nevertheless significantly improved in CSFC.
author2 School of Chemical and Biomedical Engineering
author_facet School of Chemical and Biomedical Engineering
Pu, Siyu
Hadinoto, Kunn
format Article
author Pu, Siyu
Hadinoto, Kunn
author_sort Pu, Siyu
title Comparative evaluations of bulk seeded protein crystallization in batch versus continuous slug flow crystallizers
title_short Comparative evaluations of bulk seeded protein crystallization in batch versus continuous slug flow crystallizers
title_full Comparative evaluations of bulk seeded protein crystallization in batch versus continuous slug flow crystallizers
title_fullStr Comparative evaluations of bulk seeded protein crystallization in batch versus continuous slug flow crystallizers
title_full_unstemmed Comparative evaluations of bulk seeded protein crystallization in batch versus continuous slug flow crystallizers
title_sort comparative evaluations of bulk seeded protein crystallization in batch versus continuous slug flow crystallizers
publishDate 2022
url https://hdl.handle.net/10356/160115
_version_ 1738844866976153600