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Article: Core-Shell Imprinted Particles for Adenovirus Binding.

Dietl, Sandra / Walther, Paul / Sobek, Harald / Mizaikoff, Boris

2021 Volume 14, Issue 24

Abstract: Virus-imprinted polymers were synthesized via surface imprinting strategies to produce core-shell imprinted particles selective for human adenovirus type 5. High binding affinity of the target virus towards the resulting imprinted layer was confirmed and ...

Abstract	Virus-imprinted polymers were synthesized via surface imprinting strategies to produce core-shell imprinted particles selective for human adenovirus type 5. High binding affinity of the target virus towards the resulting imprinted layer was confirmed and unspecific binding was reduced in presence of blocking agents, i.e., via bovine serum albumin and skim milk in combination with Tween 20. In addition, the imprinted materials were applied for adenovirus extraction from cell culture supernatants. High levels of virus binding with negligible binding of matrix proteins confirmed the suitability of these materials for binding and extraction of the target virus from complex matrices.
Language	English
Publishing date	2021-12-13
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2487261-1
ISSN	1996-1944
ISSN	1996-1944
DOI	10.3390/ma14247692
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Epitope-imprinted polymers for biomacromolecules: Recent strategies, future challenges and selected applications

Dietl, Sandra / Sobek, Harald / Mizaikoff, Boris

Trends in analytical chemistry. 2021 Oct., v. 143

2021

Abstract: Molecular imprinted polymers are a promising alternative to natural receptors due to their higher stability and lower production costs, yet with comparable performance and high selectivity. The ‘epitope imprinting approach’, in which only a small sub- ... ...

Abstract	Molecular imprinted polymers are a promising alternative to natural receptors due to their higher stability and lower production costs, yet with comparable performance and high selectivity. The ‘epitope imprinting approach’, in which only a small sub-unit of the target is used as the template during imprinting provides a solution to overcome several limitations characteristic for biomacromolecular imprinting including availability, complexity, size, and conformational flexibility associated with biomacromolecules. The present review summarizes recent strategies for epitope imprinting, and highlights the applicability of this approach to a wide range of targets with different sizes and molecular properties including proteins, biomarkers and viruses.
Keywords	biomarkers ; epitopes ; molecular imprinting
Language	English
Dates of publication	2021-10
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	2014041-1
ISSN	0165-9936
ISSN	0165-9936
DOI	10.1016/j.trac.2021.116414
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Article ; Online: An efficient capture strategy for the purification of human adenovirus type 5 from cell lysates.

Dietl, Sandra / Kiefer, Franziska / Binder, Stefanie / Walther, Paul / Sobek, Harald / Mizaikoff, Boris

Journal of biotechnology

2022

Abstract: An efficient capture step for human adenovirus type 5 from cell lysate was developed as an initial virus purification step from cell debris supernatant. Organosilane-based polymer particles were synthesized and experimental monomer screening allowed the ... ...

Abstract	An efficient capture step for human adenovirus type 5 from cell lysate was developed as an initial virus purification step from cell debris supernatant. Organosilane-based polymer particles were synthesized and experimental monomer screening allowed the selection of appropriate functionalities for the development of particles for virus binding. After elution, virus recoveries of 83% were obtained with significant reduction of matrix proteins and residual host cell DNA. Therefore, the implemented capture strategy for adenovirus via polymer particles provides a scalable and reproducible approach to reduce time and cost during virus purification processes.
Language	English
Publishing date	2022-12-06
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	843647-2
ISSN	1873-4863 ; 0168-1656 ; 1389-0352
ISSN (online)	1873-4863
ISSN	0168-1656 ; 1389-0352
DOI	10.1016/j.jbiotec.2022.11.015
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Article ; Online: Selective virus capture via hexon imprinting.

Gast, Manuela / Sobek, Harald / Mizaikoff, Boris

Materials science & engineering. C, Materials for biological applications

2019 Volume 99, Page(s) 1099–1104

Abstract: An imprinting technique has been developed to generate synthetic polymer beads suitable for selectively binding a supramolecular target. The viral hexon protein, which is the most abundant and accessible surface protein component of the human Adenovirus ... ...

Abstract	An imprinting technique has been developed to generate synthetic polymer beads suitable for selectively binding a supramolecular target. The viral hexon protein, which is the most abundant and accessible surface protein component of the human Adenovirus type 5 (hAdV5) icosahedral capsid, was applied as the template molecule to generate functional polymer beads entailing selectivity for the entire virus. Individual and competitive rebinding studies using two different viruses (i.e. hAdV5 and Minute Virus of Mice - MVM) revealed exquisite selectivity of the imprinted beads for the target hAdV5. Additionally, the morphology of thus imprinted beads was checked via scanning electron microscopy (SEM).
MeSH term(s)	Adenoviruses, Human/isolation & purification ; Capsid Proteins/chemistry ; Capsid Proteins/metabolism ; Humans ; Kinetics ; Molecular Imprinting/methods ; Polymers/chemistry ; Protein Binding ; Serum Albumin, Bovine/metabolism
Chemical Substances	Capsid Proteins ; Polymers ; hexon capsid protein, Adenovirus ; Serum Albumin, Bovine (27432CM55Q)
Language	English
Publishing date	2019-02-14
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	2012160-X
ISSN	1873-0191 ; 0928-4931
ISSN (online)	1873-0191
ISSN	0928-4931
DOI	10.1016/j.msec.2019.02.037
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Article ; Online: Nanoparticle Tracking of Adenovirus by Light Scattering and Fluorescence Detection.

Gast, Manuela / Sobek, Harald / Mizaikoff, Boris

Human gene therapy methods

2019 Volume 30, Issue 6, Page(s) 235–244

Abstract: The detailed characterization of biological nanoparticles is of paramount importance for various industrial sectors, as for production of viral therapeutics. More recently, technologies that allow real-time quantification with simultaneous sizing and ... ...

Abstract	The detailed characterization of biological nanoparticles is of paramount importance for various industrial sectors, as for production of viral therapeutics. More recently, technologies that allow real-time quantification with simultaneous sizing and determination of surface potentials of virus particles in solution have been developed. In this study, nanoparticle tracking analysis (NTA) was applied to determine the size and the zeta potential of human adenovirus type 5 (AdV5), one the most frequently used therapeutic/oncolytic agents and viral vectors. Virus aggregation was detected, and the kinetics of the dissolution of virus aggregates were studied in real time. In addition, advanced fluorescence detection of AdV5 was performed enabling the measurements in matrices and discrimination of viral subpopulations. It was shown that NTA is an efficient approach for investigating infectious viruses in a live viewing mode. Consequently, NTA provides a promising methodology for virus particle detection and analysis in real time beyond assays requiring nucleic acids or infectivity.
MeSH term(s)	A549 Cells ; Adenoviridae/chemistry ; Adenoviridae/isolation & purification ; Dynamic Light Scattering ; Fluoresceins/chemistry ; Humans ; Nanoparticles/chemistry ; Spectrometry, Fluorescence
Chemical Substances	Atto 488 maleimide ; Fluoresceins
Language	English
Publishing date	2019-11-21
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2654800-8
ISSN	1946-6544 ; 1946-6536
ISSN (online)	1946-6544
ISSN	1946-6536
DOI	10.1089/hgtb.2019.172
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Article ; Online: Towards the direct detection of viral materials at the surface of protective face masks via infrared spectroscopy.

Schorer, Vanessa / Haas, Julian / Stach, Robert / Kokoric, Vjekoslav / Groß, Rüdiger / Muench, Jan / Hummel, Tim / Sobek, Harald / Mennig, Jan / Mizaikoff, Boris

Scientific reports

2022 Volume 12, Issue 1, Page(s) 2309

Abstract: The ongoing COVID-19 pandemic represents a considerable risk for the general public and especially for health care workers. To avoid an overloading of the health care system and to control transmission chains, the development of rapid and cost-effective ... ...

Abstract	The ongoing COVID-19 pandemic represents a considerable risk for the general public and especially for health care workers. To avoid an overloading of the health care system and to control transmission chains, the development of rapid and cost-effective techniques allowing for the reliable diagnosis of individuals with acute respiratory infections are crucial. Uniquely, the present study focuses on the development of a direct face mask sampling approach, as worn (i.e., used) disposable face masks contain exogenous environmental constituents, as well as endogenously exhaled breath aerosols. Optical techniques-and specifically infrared (IR) molecular spectroscopic techniques-are promising tools for direct virus detection at the surface of such masks. In the present study, a rapid and non-destructive approach for monitoring exposure scenarios via medical face masks using attenuated total reflection infrared spectroscopy is presented. Complementarily, IR external reflection spectroscopy was evaluated in comparison for rapid mask analysis. The utility of a face mask-based sampling approach was demonstrated by differentiating water, proteins, and virus-like particles sampled onto the mask. Data analysis using multivariate statistical algorithms enabled unambiguously classifying spectral signatures of individual components and biospecies. This approach has the potential to be extended towards the rapid detection of SARS-CoV-2-as shown herein for the example of virus-like particles which are morphologically equivalent to authentic virus-without any additional sample preparation or elaborate testing equipment at laboratory facilities. Therefore, this strategy may be implemented as a routine large-scale monitoring routine, e.g., at health care institutions, nursing homes, etc. ensuring the health and safety of medical personnel.
MeSH term(s)	Masks/virology ; SARS-CoV-2/isolation & purification ; Spectrophotometry, Infrared
Language	English
Publishing date	2022-02-10
Publishing country	England
Document type	Evaluation Study ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-022-06335-z
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Article: Use of Super-Resolution Optical Microscopy To Reveal Direct Virus Binding at Hybrid Core–Shell Matrixes

Gast, Manuela / Wondany, Fanny / Raabe, Bastian / Michaelis, Jens / Sobek, Harald / Mizaikoff, Boris

Analytical chemistry. 2020 Jan. 06, v. 92, no. 4

2020

Abstract: Polymer particles with antibody-like affinity, i.e., molecularly imprinted polymers, offer an ideal platform for biopharmaceutical virus purification. In recent years, attempts combining molecular imprinting technology with a variety of visualization and ...

Abstract	Polymer particles with antibody-like affinity, i.e., molecularly imprinted polymers, offer an ideal platform for biopharmaceutical virus purification. In recent years, attempts combining molecular imprinting technology with a variety of visualization and detection techniques have been reported for directly confirming the localized presence of the template. Direct target visualization is crucial for the characterization of molecularly imprinted polymers, especially if biological templates such as viruses are used. In the present study, for the first time the viral binding behavior at virus-imprinted polymers (VIPs) via stimulated emission depletion (STED) microscopy is shown by imaging individual, fluorescently labeled virus particles. STED microscopy achieves among various other super-resolution techniques the best temporal resolution at high spatial resolution. An innovative virus purification material selective for human adenovirus type 5 (AdV5) offered highly purified virus for the subsequent fluorescent labeling procedure, thus enabling STED imaging. Excellent binding affinities (150-fold higher versus control particles) and high selectivity toward the target virus (AdV5) were observed at those VIPs, even in competitive binding experiments with minute virus of mice using dual-label STED microscopy.
Keywords	Human mastadenovirus C ; Rodent protoparvovirus 1 ; animal viruses ; binding capacity ; biopharmaceuticals ; fluorescent labeling ; image analysis ; light microscopy ; molecular imprinting ; polymers ; virion ; viruses
Language	English
Dates of publication	2020-0106
Size	p. 3050-3057.
Publishing place	American Chemical Society
Document type	Article
ZDB-ID	1508-8
ISSN	1520-6882 ; 0003-2700
ISSN (online)	1520-6882
ISSN	0003-2700
DOI	10.1021/acs.analchem.9b04328
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Article ; Online: Use of Super-Resolution Optical Microscopy To Reveal Direct Virus Binding at Hybrid Core-Shell Matrixes.

Gast, Manuela / Wondany, Fanny / Raabe, Bastian / Michaelis, Jens / Sobek, Harald / Mizaikoff, Boris

Analytical chemistry

2020 Volume 92, Issue 4, Page(s) 3050–3057

Abstract	Polymer particles with antibody-like affinity, i.e., molecularly imprinted polymers, offer an ideal platform for biopharmaceutical virus purification. In recent years, attempts combining molecular imprinting technology with a variety of visualization and detection techniques have been reported for directly confirming the localized presence of the template. Direct target visualization is crucial for the characterization of molecularly imprinted polymers, especially if biological templates such as viruses are used. In the present study, for the first time the viral binding behavior at virus-imprinted polymers (VIPs) via stimulated emission depletion (STED) microscopy is shown by imaging individual, fluorescently labeled virus particles. STED microscopy achieves among various other super-resolution techniques the best temporal resolution at high spatial resolution. An innovative virus purification material selective for human adenovirus type 5 (AdV5) offered highly purified virus for the subsequent fluorescent labeling procedure, thus enabling STED imaging. Excellent binding affinities (150-fold higher versus control particles) and high selectivity toward the target virus (AdV5) were observed at those VIPs, even in competitive binding experiments with minute virus of mice using dual-label STED microscopy.
MeSH term(s)	Adenoviruses, Human/chemistry ; Humans ; Microscopy, Fluorescence ; Molecular Imprinting ; Polymers/chemistry ; Virus Attachment
Chemical Substances	Polymers
Language	English
Publishing date	2020-01-16
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1508-8
ISSN	1520-6882 ; 0003-2700
ISSN (online)	1520-6882
ISSN	0003-2700
DOI	10.1021/acs.analchem.9b04328
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Article ; Online: Enhanced Selectivity by Passivation: Molecular Imprints for Viruses with Exceptional Binding Properties.

Gast, Manuela / Kühner, Stefanie / Sobek, Harald / Walther, Paul / Mizaikoff, Boris

Analytical chemistry

2018 Volume 90, Issue 9, Page(s) 5576–5585

Abstract: Inspired by the recognition processes found in biology such as enzyme-substrate and antibody-antigen interactions, synthetic systems with comparable molecular recognition properties have been investigated during recent years based on molecular imprinting ...

Abstract	Inspired by the recognition processes found in biology such as enzyme-substrate and antibody-antigen interactions, synthetic systems with comparable molecular recognition properties have been investigated during recent years based on molecular imprinting strategies. While materials with recognition capabilities for small molecules (i.e., with low molecular weight) have achieved substantial advancements, the synthesis of molecularly imprinted materials with virus recognition properties remains challenging to date. Likewise, protein-surface and protein-protein interactions are essential for a wide variety of biological applications in biotechnology. In biological sensor technology the coating of surfaces to prevent nonspecific adsorption interactions plays an important role. Particularly, polyethylene glycol (PEG) stands out for its high performance in preventing proteins from nonspecifically interactions. However, blocking agents such as the protein bovine serum albumin (BSA) can also be useful as unspecific binding prevention agents for passivation, without modification of the surface. Herein the influence of blocking agents as unspecific reaction components is investigated on the enhancements of selectivity from adenovirus-imprinted particles, whereas adenovirus was used as target species in molecular imprinting. Furthermore, quantitative polymerase chain reaction (qPCR) was used for the first time as virus quantification approach in this context.
MeSH term(s)	Adenoviridae/genetics ; Adenoviridae/isolation & purification ; Molecular Imprinting ; Molecular Structure ; Silicon Dioxide/chemical synthesis ; Silicon Dioxide/chemistry
Chemical Substances	Silicon Dioxide (7631-86-9)
Language	English
Publishing date	2018-03-19
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1508-8
ISSN	1520-6882 ; 0003-2700
ISSN (online)	1520-6882
ISSN	0003-2700
DOI	10.1021/acs.analchem.7b05148
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Article: Plastic Antibodies Mimicking the ACE2 Receptor for Selective Binding of SARS-CoV-2 Spike.

Batista, Alex D / Rajpal, Soumya / Keitel, Benedikt / Dietl, Sandra / Fresco-Cala, Beatriz / Dinc, Mehmet / Groß, Rüdiger / Sobek, Harald / Münch, Jan / Mizaikoff, Boris

Advanced materials interfaces

2022 Volume 9, Issue 5, Page(s) 2101925

Abstract: Molecular imprinting has proven to be a versatile and simple strategy to obtain selective materials also termed "plastic antibodies" for a wide variety of species, i.e., from ions to macromolecules and viruses. However, to the best of the authors' ... ...

Abstract	Molecular imprinting has proven to be a versatile and simple strategy to obtain selective materials also termed "plastic antibodies" for a wide variety of species, i.e., from ions to macromolecules and viruses. However, to the best of the authors' knowledge, the development of epitope-imprinted polymers for selective binding of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is not reported to date. An epitope from the SARS-CoV-2 spike protein comprising 17 amino acids is used as a template during the imprinting process. The interactions between the epitope template and organosilane monomers used for the polymer synthesis are predicted via molecular docking simulations. The molecularly imprinted polymer presents a 1.8-fold higher selectivity against the target epitope compared to non-imprinted control polymers. Rebinding studies with pseudoviruses containing SARS-CoV-2 spike protein demonstrate the superior selectivity of the molecularly imprinted matrices, which mimic the interactions of angiotensin-converting enzyme 2 receptors from human cells. The obtained results highlight the potential of SARS-CoV-2 molecularly imprinted polymers for a variety of applications including chem/biosensing and antiviral delivery.
Language	English
Publishing date	2022-01-05
Publishing country	Germany
Document type	Journal Article
ISSN	2196-7350
ISSN	2196-7350
DOI	10.1002/admi.202101925
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