Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
4.7
3.5
Journals
Processes
Special Issues
Recent Advances in Protein Separation and Purification Methods
share announcement

Recent Advances in Protein Separation and Purification Methods

A special issue of Processes (ISSN 2227-9717). This special issue belongs to the section "Biological Processes and Systems".

Deadline for manuscript submissions: closed (30 September 2022) | Viewed by 11405

Special Issue Editor

Prof. Dr. Yesong Gu

E-Mail Website
Guest Editor
Department of Chemical and Materials Engineering, Tunghai University, Taichung City 40704, Taiwan
Interests: protein purification process; green chemical process design for biocompatible and degradable polymers; biosensor fabrication for clinical diagnosis; genetic engineering and protein drug development; cancer research

Special Issue Information

Dear Colleagues,

Recombinant DNA technology has contributed greatly to modern biotechnology, which has enabled us to introduce various human genes into bacteria, yeast, and other mammalian cells in order to produce prospective recombinant proteins with potential diagnostic and therapeutic applications. In comparison to chemical synthetic drugs, therapeutic protein drugs may have the advantages of biocompatibility, biosafety, effectiveness, and reduced side effects. Currently, the therapeutic peptide and protein drugs have become more competitive in the entire pharmaceutical market and are a major new class of therapeutics approved for clinical use by the US-FDA. However, therapeutic proteins are not easily manufactured since they are genetically produced in the biological cells that are often under the situation of extremely low concentration and high complexity; therefore, a successful separation and purification process becomes especially important. Due to there being strict demands on the expected biological activity, extremely high purity, and biosafety, a therapeutic protein drug relies on a highly controlled biological process—in particular, the state-of-the-art design of protein purification process development.

This Special Issue on “Recent Advances on Protein Separation and Purification Methods” aims to explore the innovative advances in the development and application of modern bioseparation and purification techniques, process design optimization, as well as artificial intelligence methods in successful protein purification process design. Topics include but are not limited to:

  • Modern techniques for the protein separation and purification;
  • Concepts and methodologies in optimizing protein separation and purification process design;
  • Applications of artificial intelligence in protein process development;

Prof. Dr. Yesong Gu
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Processes is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2400 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • proteins
  • purification techniques
  • purification process design
  • process optimization

Published Papers (4 papers)

Download All Papers
Order results
Result details
Show export options expand_more Show export options expand_less
Select all
Export citation of selected articles as:

Research

16 pages, 2274 KiB  
Article
Purification of High-Molecular-Weight Antibacterial Proteins of Insect Pathogenic Brevibacillus laterosporus Isolates
by Tauseef K. Babar, Travis R. Glare, John G. Hampton, Mark R. H. Hurst, Josefina O. Narciso and Amy Beattie
Processes 2022, 10(10), 1932; https://0-doi-org.brum.beds.ac.uk/10.3390/pr10101932 - 25 Sep 2022
Cited by 1 | Viewed by 1577
Abstract
Brevibacillus laterosporus (Bl) is a Gram-positive and spore-forming bacterium belonging to the Brevibacillus brevis phylogenetic cluster. Globally, insect pathogenic strains of the bacterium have been isolated, characterised, and some activities have been patented. Two isolates, Bl 1821L and Bl 1951, exhibiting [...] Read more.
Brevibacillus laterosporus (Bl) is a Gram-positive and spore-forming bacterium belonging to the Brevibacillus brevis phylogenetic cluster. Globally, insect pathogenic strains of the bacterium have been isolated, characterised, and some activities have been patented. Two isolates, Bl 1821L and Bl 1951, exhibiting pathogenicity against the diamondback moth and mosquitoes, are under development as a biopesticide in New Zealand. However, due to the suspected activity of putative antibacterial proteins (ABPs), the endemic isolates often grow erratically. Various purification methods, including size exclusion chromatography, sucrose density gradient centrifugation, polyethylene glycol precipitation, and ammonium sulphate precipitation employed in this study, enabled the isolation of two putative antibacterial proteins of ~30 and ~48 kD from Bl 1821L and one putative antibacterial protein of ~30 kD from Bl 1951. Purification of the uninduced cultures of Bl 1821L and Bl 1951 also yielded protein bands of ~30 and ~48 kD on SDS-PAGE, which indicated their spontaneous induction. A disc diffusion assay was used to determine the antagonistic activities of the putative ABPs. Subsequent transmission electron microscope (TEM) examination of a purified putative antibacterial protein-containing solution showed the presence of encapsulin (~30 kD) and polysheath (~48 kD)-like structures. Although only the ~30 kD protein was purified from Bl 1951, both structures were seen in this strain under TEM. Furthermore, while assessing the antibacterial activity of some fractions of Bl 1951 against Bl 1821L in the size exclusion chromatography method, the population of Bl 1821L persister cells was noted. Overall, this work added a wealth of knowledge about the purification of the high-molecular-weight (HMW) proteins (bacteriocins) of Gram-positive bacteria including Bl. Full article
(This article belongs to the Special Issue Recent Advances in Protein Separation and Purification Methods)
►▼ Show Figures

Graphical abstract

18 pages, 2805 KiB  
Article
An Optimized Purification Design for Extracting Active ADAMTS13 from Conditioned Media
by Katarzyna I. Jankowska, Upendra Katneni, Brian C. Lin, Randilu Amarasinghe, Je-Nie Phue, Wells W. Wu, Nobuko Hamasaki-Katagiri, Wojciech Jankowski, Rong-Fong Shen and Chava Kimchi-Sarfaty
Processes 2022, 10(2), 322; https://0-doi-org.brum.beds.ac.uk/10.3390/pr10020322 - 08 Feb 2022
Cited by 1 | Viewed by 2131
Abstract
ADAMTS13 is a hemostatic enzyme that breaks down pro-thrombotic ultra-large multimers of von Willebrand factor (VWF). The deficiency of ADAMTS13 increases VWF-mediated thrombogenic potential and may lead to thrombotic thrombocytopenic purpura (TTP). Recently, clinical studies have shown the development of acquired TTP after [...] Read more.
ADAMTS13 is a hemostatic enzyme that breaks down pro-thrombotic ultra-large multimers of von Willebrand factor (VWF). The deficiency of ADAMTS13 increases VWF-mediated thrombogenic potential and may lead to thrombotic thrombocytopenic purpura (TTP). Recently, clinical studies have shown the development of acquired TTP after COVID-19 infection and a correlation between low ADAMTS13 plasma levels and increased mortality. As a result, investigating ADAMTS13 as a potential recombinant therapeutic is of broad interest in the field of hematology. ADAMTS13 is considered challenging to purify in its biologically active state. Current purification methods utilize immobilized metal ions, which can interfere with ADAMTS13 metalloprotease activity. For this reason, we optimized an alternative strategy to isolate milligram quantities of highly active recombinant ADAMTS13 (rADAMTS13) from conditioned media after exogenous expression in human cell line, HEK293. HEK293 cells stably expressing C-terminal V5-His-tagged ADAMTS13 were grown in two parallel systems, culture bottles and flasks, for identifying an optimal cultivation strategy. Subsequently, we employed anion exchange followed by anti-V5-tag affinity chromatography to purify rADAMTS13, and extracted rADAMTS13 of high specific activity while preserving its native post-translational modifications. In addition, this process has been optimized and scaled up to produce active rADAMTS13 at levels sufficient for laboratory-scale structural, enzymatic, and biochemical studies. Full article
(This article belongs to the Special Issue Recent Advances in Protein Separation and Purification Methods)
►▼ Show Figures

Figure 1

18 pages, 1961 KiB  
Article
Design of Experiment (DoE) for Optimization of HPLC Conditions for the Simultaneous Fractionation of Seven α-Amylase/Trypsin Inhibitors from Wheat (Triticum aestivum L.)
by Sorel Tchewonpi Sagu, Gerd Huschek, Tess Waldbach Braga, Michal Rackiewicz, Thomas Homann and Harshadrai M. Rawel
Processes 2022, 10(2), 259; https://0-doi-org.brum.beds.ac.uk/10.3390/pr10020259 - 28 Jan 2022
Cited by 1 | Viewed by 3161
Abstract
Wheat alpha-amylase/trypsin inhibitors remain a subject of interest considering the latest findings showing their implication in wheat-related non-celiac sensitivity (NCWS). Understanding their functions in such a disorder is still unclear and for further study, the need for pure ATI molecules is one of [...] Read more.
Wheat alpha-amylase/trypsin inhibitors remain a subject of interest considering the latest findings showing their implication in wheat-related non-celiac sensitivity (NCWS). Understanding their functions in such a disorder is still unclear and for further study, the need for pure ATI molecules is one of the limiting problems. In this work, a simplified approach based on the successive fractionation of ATI extracts by reverse phase and ion exchange chromatography was developed. ATIs were first extracted from wheat flour using a combination of Tris buffer and chloroform/methanol methods. The separation of the extracts on a C18 column generated two main fractions of interest F1 and F2. The response surface methodology with the Doehlert design allowed optimizing the operating parameters of the strong anion exchange chromatography. Finally, the seven major wheat ATIs namely P01083, P17314, P16850, P01085, P16851, P16159, and P83207 were recovered with purity levels (according to the targeted LC-MS/MS analysis) of 98.2 ± 0.7; 98.1 ± 0.8; 97.9 ± 0.5; 95.1 ± 0.8; 98.3 ± 0.4; 96.9 ± 0.5, and 96.2 ± 0.4%, respectively. MALDI-TOF-MS analysis revealed single peaks in each of the pure fractions and the mass analysis yielded deviations of 0.4, 1.9, 0.1, 0.2, 0.2, 0.9, and 0.1% between the theoretical and the determined masses of P01083, P17314, P16850, P01085, P16851, P16159, and P83207, respectively. Overall, the study allowed establishing an efficient purification process of the most important wheat ATIs. This paves the way for further in-depth investigation of the ATIs to gain more knowledge related to their involvement in NCWS disease and to allow the absolute quantification in wheat samples. Full article
(This article belongs to the Special Issue Recent Advances in Protein Separation and Purification Methods)
►▼ Show Figures

Figure 1

9 pages, 2108 KiB  
Communication
Design of Experiments for Modeling of Fermentation Process Characterization in Biological Drug Production
by Kuang-Lung Hsueh, Tzung-Yi Lin, Meng-Tse (Gabriel) Lee, Ya-Yun Hsiao and Yesong Gu
Processes 2022, 10(2), 237; https://0-doi-org.brum.beds.ac.uk/10.3390/pr10020237 - 26 Jan 2022
Cited by 2 | Viewed by 3396
Abstract
Biological products are increasingly important, and therefore the industry has begun to adopt quality by design, as recommended by the ICH and the U.S. FDA. Smaller companies, however, have faced difficulties in employing full-scale experiments or the quality by design strategy. Thus, this [...] Read more.
Biological products are increasingly important, and therefore the industry has begun to adopt quality by design, as recommended by the ICH and the U.S. FDA. Smaller companies, however, have faced difficulties in employing full-scale experiments or the quality by design strategy. Thus, this study provides an alternative way to build a model from existing data with experimental software that does not require full-scale experiments. This empirical study hopes to provide a practical way to improve the efficiency of smaller biopharmaceutical companies and researchers. Moreover, the models provided here can be applied to process characterization in recombinant protein production. Full article
(This article belongs to the Special Issue Recent Advances in Protein Separation and Purification Methods)
►▼ Show Figures

Figure 1

Show export options expand_more Show export options expand_less
Select all
Export citation of selected articles as:
 
Displaying articles 1-4
Back to TopTop