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Recent Advances in Bioseparation Technologies for the Capture of Emerging Biomacromolecules

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Macromolecules".

Deadline for manuscript submissions: closed (31 January 2025) | Viewed by 7600

Special Issue Editors

Dr. Luís Passarinha

E-Mail Website
Guest Editor
1. Associate Laboratory i4HB, Institute for Health and Bioeconomy, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2819-516 Caparica, Portugal
2. UCIBIO–Applied Molecular Biosciences Unit, Chemistry Department, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal
3. CICS-UBI-Health Sciences Research Centre, University of Beira Interior, 6201-506 Covilhã, Portugal
Interests: method development and validation; electrochemical detection; proteomics; protein biomarkers
Special Issues, Collections and Topics in MDPI journals
Dr. Ângela Sousa

E-Mail Website
Guest Editor
CICS-UBI–Health Sciences Research Centre, University of Beira Interior, 6201-506 Covilhã, Portugal
Interests: separation and purification methodologies; recombinant proteins; therapeutic nucleic acids
Special Issues, Collections and Topics in MDPI journals
Prof. Dr. Eugenia Gallardo

E-Mail Website
Guest Editor
1. Health Sciences Research Centre, University of Beira Interior (CICS-UBI), 6200-506 Covilhã, Portugal
2. Pharmaco-Toxicology Laboratory, UBIMedical, University of Beira Interior, 6200-284 Covilhã, Portugal
3. Centro Académico Clínico das Beiras (CACB)—Grupo de Problemas Relacionados com Toxicofilias, 6200-284 Covilhã, Portugal
Interests: toxicology; analytical method development; recreational drugs; natural psychoactive substances; therapeutic drug monitoring; sample preparation; alternative samples; miniaturized extraction procedures
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Several research groups are devoted to the dissemination of new and original knowledge in all fields of bioseparation sciences. In order to assemble these findings, a Special Issue entitled “Recent Advances in Bioseparation Technologies for the Capture of Emerging Biomacromolecules” is being launched. This Special Issue aims to gather scientific full papers, short communications, and review articles covering novel separation and isolation methods based on chromatographic and nonchromatographic platforms, demonstrating a huge societal impact—for instance, advances in the purification of biomacromolecules or pathological factors using microdevices and advanced aqueous two-phase systems. Developments with interdisciplinary approaches are particularly welcome, and systems should be proven with suitable biological, therapeutic, or biotechnological challenging samples.

Presenting a very broad scope, this Special Issue welcomes but is not limited to the following technologies and emergent applications: molecular diagnostics, biosensors and bioengineering, drug development and pharmaceutical analysis, microfluidics and nanotechnology, omics analysis (such as proteomics, metabolomics or glycomics), food science, neuroscience, biochemical and forensic analysis, industrial processes, and method development.

This issue will serve as a collection of papers for academia or as a reference tool for researchers, including health and industry professionals, particularly those working in the fields of biotechnology, biomedicine, biology, chemistry, drug design, medicine, and pharmaceutical sciences.

We look forward to receiving your contributions.

Dr. Luís Passarinha
Dr. Ângela Sousa
Prof. Dr. Eugenia Gallardo
Guest Editors

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. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. There is an Article Processing Charge (APC) for publication in this open access journal. For details about the APC please see here. 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

  • nucleic acids
  • proteins
  • purification
  • isolation

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Further information on MDPI's Special Issue policies can be found here.

Published Papers (3 papers)

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32 pages, 11157 KiB  
Article
Simultaneous Purification of Human Interferon Alpha-2b and Serum Albumin Using Bioprivileged Fluorinated Ionic Liquid-Based Aqueous Biphasic Systems
by Sara F. Carvalho, Ana B. Pereiro and João M. M. Araújo
Int. J. Mol. Sci. 2024, 25(5), 2751; https://doi.org/10.3390/ijms25052751 - 27 Feb 2024
Cited by 2 | Viewed by 2461
Abstract
Interferon alpha-2b (IFN-α2b) is an essential cytokine widely used in the treatment of chronic hepatitis C and hairy cell leukemia, and serum albumin is the most abundant plasma protein with numerous physiological functions. Effective single-step aqueous biphasic system (ABS) extraction for [...] Read more.
Interferon alpha-2b (IFN-α2b) is an essential cytokine widely used in the treatment of chronic hepatitis C and hairy cell leukemia, and serum albumin is the most abundant plasma protein with numerous physiological functions. Effective single-step aqueous biphasic system (ABS) extraction for the simultaneous purification of IFN-α2b and BSA (serum albumin protein) was developed in this work. Effects of the ionic liquid (IL)-based ABS functionalization, fluorinated ILs (FILs; [C​2C​1Im][C​4F​9SO​3] and [N​1112(OH)][C​4F​9SO​3]) vs. mere fluoro-containing IL ([C​4C​1Im][CF​3SO​3]), in combination with sucrose or [N​1112(OH)][H​2PO​4] (well-known globular protein stabilizers), or high-charge-density salt K​3PO​4 were investigated. The effects of phase pH, phase water content (%wt), phase composition (%wt), and phase volume ratio were investigated. The phase pH was found to have a significant effect on IFN-α2b and BSA partition. Experimental results show that simultaneous single-step purification was achieved with a high yield (extraction efficiency up to 100%) for both proteins and a purification factor of IFN-α2b high in the enriched IFN-α2b phase (up to 23.22) and low in the BSA-enriched phase (down to 0.00). SDS-PAGE analysis confirmed the purity of both recovered proteins. The stability and structure of IFN-α2b and BSA were preserved or even improved (FIL-rich phase) during the purification step, as evaluated by CD spectroscopy and DSC. Binding studies of IFN-α2b and BSA with the ABS phase-forming components were assessed by MST, showing the strong interaction between FILs aggregates and both proteins. In view of their biocompatibility, customizable properties, and selectivity, FIL-based ABSs are suggested as an improved purification step that could facilitate the development of biologics. Full article
(This article belongs to the Special Issue Recent Advances in Bioseparation Technologies for the Capture of Emerging Biomacromolecules)
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18 pages, 2208 KiB  
Article
Isomers Recognition in HPLC-MS/MS Analysis of Human Plasma Samples by Using an Ion Trap Supported by a Linear Equations-Based Algorithm
by Marco Pallecchi, Luigi Lucio, Laura Braconi, Marta Menicatti, Silvia Dei, Elisabetta Teodori and Gianluca Bartolucci
Int. J. Mol. Sci. 2023, 24(13), 11155; https://doi.org/10.3390/ijms241311155 - 6 Jul 2023
Cited by 5 | Viewed by 1710
Abstract
The tandem mass spectrometry (MS/MS) approach employing an ion trap mass analyzer (IT) was evaluated in isomers recognition. The proposed approach consists of sole, simple, and rapid liquid chromatographic separation (HPLC) without requiring resolution between the analytes. Then, the MS/MS properties were optimized [...] Read more.
The tandem mass spectrometry (MS/MS) approach employing an ion trap mass analyzer (IT) was evaluated in isomers recognition. The proposed approach consists of sole, simple, and rapid liquid chromatographic separation (HPLC) without requiring resolution between the analytes. Then, the MS/MS properties were optimized to solve the signal assignment using post-processing data elaboration (LEDA). The IT-MS/MS experiment uses the same site, helium as collision gas, and different time steps to modify the applied conditions on the studied ions. Nevertheless, helium cannot ensure the quick energization of the precursor ion due to its small cross-section. Then, different combinations between excitation amplitude (ExA) and excitation time (ExT) were tested to achieve the activation of the fragmentation channels and the formation of the MS/MS spectrum. Usually, the IT-MS/MS acquisition cycle is longer for other multistage instruments, decreasing the frequency of sample data collection and influencing the chromatographic profile. To solve these problems, two time segments were set up, and the elution conditions were optimized with a compromise between peaks distinction and run time reduction. The developed HPLC-MS/MS method was checked and applied to analyze a series of human plasma samples spiked with an equimolar mixture of pair of isomers. Full article
(This article belongs to the Special Issue Recent Advances in Bioseparation Technologies for the Capture of Emerging Biomacromolecules)
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19 pages, 3030 KiB  
Article
Specific Six-Transmembrane Epithelial Antigen of the Prostate 1 Capture with Gellan Gum Microspheres: Design, Optimization and Integration
by João Batista-Silva, Diana Gomes, Jorge Barroca-Ferreira, Eugénia Gallardo, Ângela Sousa and Luís A. Passarinha
Int. J. Mol. Sci. 2023, 24(3), 1949; https://doi.org/10.3390/ijms24031949 - 18 Jan 2023
Viewed by 2401
Abstract
This work demonstrates the potential of calcium- and nickel-crosslinked Gellan Gum (GG) microspheres to capture the Six-Transmembrane Epithelial Antigen of the Prostate 1 (STEAP1) directly from complex Komagataella pastoris mini-bioreactor lysates in a batch method. Calcium-crosslinked microspheres were applied in an ionic exchange [...] Read more.
This work demonstrates the potential of calcium- and nickel-crosslinked Gellan Gum (GG) microspheres to capture the Six-Transmembrane Epithelial Antigen of the Prostate 1 (STEAP1) directly from complex Komagataella pastoris mini-bioreactor lysates in a batch method. Calcium-crosslinked microspheres were applied in an ionic exchange strategy, by manipulation of pH and ionic strength, whereas nickel-crosslinked microspheres were applied in an affinity strategy, mirroring a standard immobilized metal affinity chromatography. Both formulations presented small diameters, with appreciable crosslinker content, but calcium-crosslinked microspheres were far smoother. The most promising results were obtained for the ionic strategy, wherein calcium-crosslinked GG microspheres were able to completely bind 0.1% (v/v) DM solubilized STEAP1 in lysate samples (~7 mg/mL). The target protein was eluted in a complexed state at pH 11 with 500 mM NaCl in 10 mM Tris buffer, in a single step with minimal losses. Coupling the batch clarified sample with a co-immunoprecipitation polishing step yields a sample of monomeric STEAP1 with a high degree of purity. For the first time, we demonstrate the potential of a gellan batch method to function as a clarification and primary capture method towards STEAP1, a membrane protein, simplifying and reducing the costs of standard purification workflows. Full article
(This article belongs to the Special Issue Recent Advances in Bioseparation Technologies for the Capture of Emerging Biomacromolecules)
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