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Article ; Online: Vaccine Adjuvants Differentially Affect Kinetics of Antibody and Germinal Center Responses.

Pedersen, Gabriel Kristian / Wørzner, Katharina / Andersen, Peter / Christensen, Dennis

2020 Volume 11, Page(s) 579761

Abstract: Aluminum salts and squalene based oil-in-water emulsions (SE) are widely used adjuvants in licensed vaccines, yet their mechanisms are not fully known. Here we report that induction of antibody responses displays different kinetics dependent on the ... ...

Abstract	Aluminum salts and squalene based oil-in-water emulsions (SE) are widely used adjuvants in licensed vaccines, yet their mechanisms are not fully known. Here we report that induction of antibody responses displays different kinetics dependent on the adjuvant used. SE facilitated a rapid antibody response in contrast to aluminum hydroxide (AH) and the depot-forming cationic liposome-based adjuvant (CAF01). Antigen given with the SE adjuvant rapidly reached follicular B cells in the draining lymph node, whereas antigen formulated in AH or CAF01 remained at the site of injection as a depot. Removal of the injection site early after immunization abrogated antibody responses only when antigen was given in the depot-forming adjuvants. Despite initial delays in B cell activation and germinal center (GC) formation when antigen was given in depot-forming adjuvants, the antibody levels reached higher magnitudes than when the antigen was formulated in SE. This study demonstrates that the kinetic aspect of antibody responses is critical in adjuvant benchmarking and suggests that the optimal vaccination regime depends on the adjuvant used.
MeSH term(s)	Adjuvants, Immunologic/administration & dosage ; Adjuvants, Immunologic/chemistry ; Aluminum/chemistry ; Animals ; Antibodies/metabolism ; Antibody Formation ; B-Lymphocytes/immunology ; Female ; Germinal Center/immunology ; Liposomes/administration & dosage ; Lymphocyte Activation ; Mice ; Squalene/chemistry ; Vaccination ; Vaccines
Chemical Substances	Adjuvants, Immunologic ; Antibodies ; Liposomes ; Vaccines ; Squalene (7QWM220FJH) ; Aluminum (CPD4NFA903)
Language	English
Publishing date	2020-09-23
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2020.579761
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Article ; Online: Author Correction: Co-adjuvanting DDA/TDB liposomes with a TLR7 agonist allows for IgG2a/c class-switching in the absence of Th1 cells.

Zimmermann, Julie / van Haren, Simon D / Diray-Arce, Joann / Adriawan, Ignatius Ryan / Wørzner, Katharina / Krog, Ricki T / Guleed, Safia / Hu, Tu / Mortensen, Rasmus / Dietrich, Jes / Solbak, Sara M Ø / Levy, Ofer / Christensen, Dennis / Pedersen, Gabriel K

NPJ vaccines

2024 Volume 9, Issue 1, Page(s) 13

Language	English
Publishing date	2024-01-11
Publishing country	England
Document type	Published Erratum
ISSN	2059-0105
ISSN (online)	2059-0105
DOI	10.1038/s41541-024-00804-4
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Article ; Online: Repeated immunization with ATRA-containing liposomal adjuvant transdifferentiates Th17 cells to a Tr1-like phenotype.

Wørzner, Katharina / Zimmermann, Julie / Buhl, Regitze / Desoi, Anna / Christensen, Dennis / Dietrich, Jes / Nguyen, Nina Dieu Nhien Tran / Lindenstrøm, Thomas / Woodworth, Joshua S / Alhakeem, Reham Sabah / Yu, Steven / Ødum, Niels / Mortensen, Rasmus / Ashouri, Judith F / Pedersen, Gabriel K

Journal of autoimmunity

2024 Volume 144, Page(s) 103174

Abstract: In many autoimmune diseases, autoantigen-specific Th17 cells play a pivotal role in disease pathogenesis. Th17 cells can transdifferentiate into other T cell subsets in inflammatory conditions, however, there have been no attempts to target Th17 cell ... ...

Abstract	In many autoimmune diseases, autoantigen-specific Th17 cells play a pivotal role in disease pathogenesis. Th17 cells can transdifferentiate into other T cell subsets in inflammatory conditions, however, there have been no attempts to target Th17 cell plasticity using vaccines. We investigated if autoantigen-specific Th17 cells could be specifically targeted using a therapeutic vaccine approach, where antigen was formulated in all-trans retinoic acid (ATRA)-containing liposomes, permitting co-delivery of antigen and ATRA to the same target cell. Whilst ATRA was previously found to broadly reduce Th17 responses, we found that antigen formulated in ATRA-containing cationic liposomes only inhibited Th17 cells in an antigen-specific manner and not when combined with an irrelevant antigen. Furthermore, this approach shifted existing Th17 cells away from IL-17A expression and transcriptomic analysis of sorted Th17 lineage cells from IL-17 fate reporter mice revealed a shift of antigen-specific Th17 cells to exTh17 cells, expressing functional markers associated with T cell regulation and tolerance. In the experimental autoimmune encephalomyelitis (EAE) mouse model of MS, vaccination with myelin-specific (MOG) antigen in ATRA-containing liposomes reduced Th17 responses and alleviated disease. This highlights the potential of therapeutic vaccination for changing the phenotype of existing Th17 cells in the context of immune mediated diseases.
MeSH term(s)	Mice ; Animals ; Th17 Cells ; Liposomes/metabolism ; Tretinoin/pharmacology ; Tretinoin/metabolism ; Encephalomyelitis, Autoimmune, Experimental ; Autoantigens/metabolism ; Adjuvants, Immunologic ; Immunization ; Vaccination ; Phenotype ; Mice, Inbred C57BL ; Th1 Cells
Chemical Substances	Liposomes ; Tretinoin (5688UTC01R) ; Autoantigens ; Adjuvants, Immunologic
Language	English
Publishing date	2024-02-19
Publishing country	England
Document type	Journal Article
ZDB-ID	639452-8
ISSN	1095-9157 ; 0896-8411
ISSN (online)	1095-9157
ISSN	0896-8411
DOI	10.1016/j.jaut.2024.103174
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Article ; Online: Adsorption of protein antigen to the cationic liposome adjuvant CAF®01 is required for induction of Th1 and Th17 responses but not for antibody induction.

Wørzner, Katharina / Hvannastein, Jóhanna / Schmidt, Signe Tandrup / Foged, Camilla / Rosenkrands, Ida / Pedersen, Gabriel Kristian / Christensen, Dennis

European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V

2021 Volume 165, Page(s) 293–305

Abstract: The degree of antigen adsorption to adjuvants in subunit vaccines may significantly influence the immune responses they induce upon vaccination. Commonly used approaches for studying how the level of adsorption affects the induction of antigen-specific ... ...

Abstract	The degree of antigen adsorption to adjuvants in subunit vaccines may significantly influence the immune responses they induce upon vaccination. Commonly used approaches for studying how the level of adsorption affects the induction of antigen-specific immune responses include (i) using adjuvants with different abilities to adsorb antigens, and (ii) comparing different antigens selected based on their ability to adsorb to the adjuvant. A weakness of these approaches is that not only the antigen adsorption level is varied, but also other important functional factors such as adjuvant composition and/or the B/T cell epitopes, which may affect immunogenicity. Hence, we investigated how changing the adsorption capabilities of a single antigen to an adjuvant influenced the vaccine-induced immune responses. The model antigen lysozyme, which displays a positive net charge at physiological pH due to an isoelectric point (pI) of 11, was succinylated to different extents, resulting in a reduction of the pI value to 4.4-5.9, depending on the degree of succinylation. A pronounced inverse correlation was found between the pI value of the succinylated lysozyme analogues and the degree of adsorption to a cationic liposomal adjuvant consisting of dimethyldioctadecylammonium bromide (DDA) and trehalose dibehenate (TDB) (CAF®01). Furthermore, increased adsorption to this adjuvant correlated directly with the magnitude of lysozyme-specific Th1/Th17 immune responses induced by the vaccine in mice, while there was an inverse correlation with antibody induction. However, high lysozyme-specific antibody titers were induced with an increased antigen dose, even upon vaccination with a strongly adsorbed succinylated lysozyme analogue. Hence, these data illustrate that the degree of lysozyme adsorption to CAF®01 strongly affects the quality of the resulting immune responses.
MeSH term(s)	Adjuvants, Immunologic/administration & dosage ; Adjuvants, Immunologic/chemistry ; Adsorption ; Animals ; Antigens/administration & dosage ; Antigens/chemistry ; Antigens/immunology ; Cations/administration & dosage ; Cations/chemistry ; Female ; Glycolipids/administration & dosage ; Glycolipids/chemistry ; Immunogenicity, Vaccine ; Liposomes ; Mice ; Models, Animal ; Muramidase/administration & dosage ; Muramidase/chemistry ; Muramidase/immunology ; Quaternary Ammonium Compounds/administration & dosage ; Quaternary Ammonium Compounds/chemistry ; Th1 Cells ; Th17 Cells ; Vaccines, Subunit/administration & dosage ; Vaccines, Subunit/chemistry ; Vaccines, Subunit/immunology
Chemical Substances	Adjuvants, Immunologic ; Antigens ; Cations ; Glycolipids ; Liposomes ; Quaternary Ammonium Compounds ; Vaccines, Subunit ; trehalose 6,6'-dibehenate ; dimethyldioctadecylammonium (251IW5I21C) ; Muramidase (EC 3.2.1.17)
Language	English
Publishing date	2021-05-24
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	1065368-5
ISSN	1873-3441 ; 0939-6411
ISSN (online)	1873-3441
ISSN	0939-6411
DOI	10.1016/j.ejpb.2021.05.020
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Article ; Online: Co-adjuvanting DDA/TDB liposomes with a TLR7 agonist allows for IgG2a/c class-switching in the absence of Th1 cells.

NPJ vaccines

2023 Volume 8, Issue 1, Page(s) 189

Abstract: Class-switching to IgG2a/c in mice is a hallmark response to intracellular pathogens. T cells can promote class-switching and the predominant pathway for induction of IgG2a/c antibody responses has been suggested to be via stimulation from Th1 cells. We ... ...

Abstract	Class-switching to IgG2a/c in mice is a hallmark response to intracellular pathogens. T cells can promote class-switching and the predominant pathway for induction of IgG2a/c antibody responses has been suggested to be via stimulation from Th1 cells. We previously formulated CAF®01 (cationic liposomes containing dimethyldioctadecylammonium bromide (DDA) and Trehalose-6,6-dibehenate (TDB)) with the lipidated TLR7/8 agonist 3M-052 (DDA/TDB/3M-052), which promoted robust Th1 immunity in newborn mice. When testing this adjuvant in adult mice using the recombinant Chlamydia trachomatis (C.t.) vaccine antigen CTH522, it similarly enhanced IgG2a/c responses compared to DDA/TDB, but surprisingly reduced the magnitude of the IFN-γ+Th1 response in a TLR7 agonist dose-dependent manner. Single-cell RNA-sequencing revealed that DDA/TDB/3M-052 liposomes initiated early transcription of class-switch regulating genes directly in pre-germinal center B cells. Mixed bone marrow chimeras further demonstrated that this adjuvant did not require Th1 cells for IgG2a/c switching, but rather facilitated TLR7-dependent T-bet programming directly in B cells. This study underlines that adjuvant-directed IgG2a/c class-switching in vivo can occur in the absence of T-cell help, via direct activation of TLR7 on B cells and positions DDA/TDB/3M-052 as a powerful adjuvant capable of eliciting type I-like immunity in B cells without strong induction of Th1 responses.
Language	English
Publishing date	2023-12-22
Publishing country	England
Document type	Journal Article
ISSN	2059-0105
ISSN (online)	2059-0105
DOI	10.1038/s41541-023-00781-0
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Article ; Online: Adjuvanted SARS-CoV-2 spike protein elicits neutralizing antibodies and CD4 T cell responses after a single immunization in mice.

Wørzner, Katharina / Sheward, Daniel J / Schmidt, Signe Tandrup / Hanke, Leo / Zimmermann, Julie / McInerney, Gerald / Karlsson Hedestam, Gunilla B / Murrell, Ben / Christensen, Dennis / Pedersen, Gabriel Kristian

EBioMedicine

2021 Volume 63, Page(s) 103197

Abstract: Background: SARS-CoV-2 has caused a global pandemic, infecting millions of people. A safe, effective vaccine is urgently needed and remains a global health priority. Subunit vaccines are used successfully against other viruses when administered in the ... ...

Abstract	Background: SARS-CoV-2 has caused a global pandemic, infecting millions of people. A safe, effective vaccine is urgently needed and remains a global health priority. Subunit vaccines are used successfully against other viruses when administered in the presence of an effective adjuvant. Methods: We evaluated three different clinically tested adjuvant systems in combination with the SARS-CoV-2 pre-fusion stabilized (S-2P) spike protein using a one-dose regimen in mice. Findings: Whilst spike protein alone was only weakly immunogenic, the addition of either Aluminum hydroxide, a squalene based oil-in-water emulsion system (SE) or a cationic liposome-based adjuvant significantly enhanced antibody responses against the spike receptor binding domain (RBD). Kinetics of antibody responses differed, with SE providing the most rapid response. Neutralizing antibodies developed after a single immunization in all adjuvanted groups with ID Interpretation: These results demonstrate that adjuvanted spike protein subunit vaccine is a viable strategy for rapidly eliciting SARS-CoV-2 neutralizing antibodies and CD4 T cell responses of various qualities depending on the adjuvant used, which can be explored in further vaccine development against COVID-19. Funding: This work was supported by the European Union Horizon 2020 research and innovation program under grant agreement no. 101003653.
MeSH term(s)	Adjuvants, Immunologic/administration & dosage ; Adjuvants, Immunologic/chemistry ; Aluminum Hydroxide/chemistry ; Animals ; Antibodies, Neutralizing/immunology ; Antibodies, Neutralizing/metabolism ; Antibodies, Viral/immunology ; Antibodies, Viral/metabolism ; CD4-Positive T-Lymphocytes/cytology ; CD4-Positive T-Lymphocytes/immunology ; CD4-Positive T-Lymphocytes/metabolism ; COVID-19/pathology ; COVID-19/virology ; Female ; Immunization ; Interferon-gamma/metabolism ; Interleukin-17/metabolism ; Liposomes/chemistry ; Mice ; Mice, Inbred C57BL ; SARS-CoV-2/isolation & purification ; SARS-CoV-2/metabolism ; Spike Glycoprotein, Coronavirus/chemistry ; Spike Glycoprotein, Coronavirus/immunology ; Squalene/chemistry ; Vaccines, Subunit/immunology
Chemical Substances	Adjuvants, Immunologic ; Antibodies, Neutralizing ; Antibodies, Viral ; Interleukin-17 ; Liposomes ; Spike Glycoprotein, Coronavirus ; Vaccines, Subunit ; spike protein, SARS-CoV-2 ; Aluminum Hydroxide (5QB0T2IUN0) ; Squalene (7QWM220FJH) ; Interferon-gamma (82115-62-6)
Language	English
Publishing date	2021-01-07
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	2851331-9
ISSN	2352-3964
ISSN (online)	2352-3964
DOI	10.1016/j.ebiom.2020.103197
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Article ; Online: Comparison of two different PEGylation strategies for the liposomal adjuvant CAF09: Towards induction of CTL responses upon subcutaneous vaccine administration.

Schmidt, Signe Tandrup / Olsen, Camilla Line / Franzyk, Henrik / Wørzner, Katharina / Korsholm, Karen Smith / Rades, Thomas / Andersen, Peter / Foged, Camilla / Christensen, Dennis

European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V

2019 Volume 140, Page(s) 29–39

Abstract: Using subunit vaccines, e.g., based on peptide or protein antigens, to teach the immune system to kill abnormal host cells via induction of cytotoxic T lymphocytes (CTL) is a promising strategy against intracellular infections and cancer. However, ... ...

Abstract	Using subunit vaccines, e.g., based on peptide or protein antigens, to teach the immune system to kill abnormal host cells via induction of cytotoxic T lymphocytes (CTL) is a promising strategy against intracellular infections and cancer. However, customized adjuvants are required to potentiate antigen-specific cellular immunity. One strong CTL-inducing adjuvant is the liposomal cationic adjuvant formulation (CAF)09, which is composed of dimethyldioctadecylammonium (DDA) bromide, monomycoloyl glycerol (MMG) analogue 1 and polyinosinic:polycytidylic acid [poly(I:C)]. However, this strong CTL induction requires intraperitoneal administration because the vaccine forms a depot at the site of injection (SOI) after subcutaneous (s.c.) or intramuscular (i.m.) injection, and depot formation impedes the crucial vaccine targeting to the cross-presenting dendritic cells (DCs) residing in the lymph nodes (LNs). The purpose of the present study was to investigate the effect of polyethylene glycol (PEG) grafting of CAF09 on the ability of the vaccine to induce antigen-specific CTL responses after s.c. administration. We hypothesized that steric stabilization and charge shielding of CAF09 by PEGylation may reduce depot formation at the SOI and enhance passive drainage to the LNs, eventually improving CTL induction. Hence, the vaccine (antigen/CAF09) was post-grafted with a novel type of anionic PEGylated peptides based on GDGDY repeats, which were end-conjugated with one or two PEG
MeSH term(s)	Adjuvants, Immunologic/chemistry ; Adjuvants, Immunologic/pharmacology ; Animals ; Antigens/immunology ; CD8-Positive T-Lymphocytes/immunology ; Cross-Priming/immunology ; Dendritic Cells/immunology ; Female ; Immunity, Cellular/immunology ; Immunization/methods ; Liposomes/chemistry ; Mice ; Mice, Inbred C57BL ; Ovalbumin/immunology ; Phosphatidylethanolamines/chemistry ; Polyethylene Glycols/chemistry ; Quaternary Ammonium Compounds/chemistry ; T-Lymphocytes, Cytotoxic/immunology ; Tissue Distribution ; Vaccines, Subunit/immunology
Chemical Substances	Adjuvants, Immunologic ; Antigens ; Liposomes ; Phosphatidylethanolamines ; Quaternary Ammonium Compounds ; Vaccines, Subunit ; dimethyldioctadecylammonium (251IW5I21C) ; Polyethylene Glycols (3WJQ0SDW1A) ; Ovalbumin (9006-59-1) ; polyethylene glycol 2000 (HAF0412YIT) ; polyethylene glycol 1000 (U076Q6Q621)
Language	English
Publishing date	2019-05-02
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	1065368-5
ISSN	1873-3441 ; 0939-6411
ISSN (online)	1873-3441
ISSN	0939-6411
DOI	10.1016/j.ejpb.2019.04.020
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Article ; Online: Adjuvanted SARS-CoV-2 spike protein elicits neutralizing antibodies and CD4 T cell responses after a single immunization in mice

Katharina Wørzner / Daniel J. Sheward / Signe Tandrup Schmidt / Leo Hanke / Julie Zimmermann / Gerald McInerney / Gunilla B. Karlsson Hedestam / Ben Murrell / Dennis Christensen / Gabriel Kristian Pedersen

EBioMedicine, Vol 63, Iss , Pp 103197- (2021)

2021

Abstract	Background: SARS-CoV-2 has caused a global pandemic, infecting millions of people. A safe, effective vaccine is urgently needed and remains a global health priority. Subunit vaccines are used successfully against other viruses when administered in the presence of an effective adjuvant. Methods: We evaluated three different clinically tested adjuvant systems in combination with the SARS-CoV-2 pre-fusion stabilized (S-2P) spike protein using a one-dose regimen in mice. Findings: Whilst spike protein alone was only weakly immunogenic, the addition of either Aluminum hydroxide, a squalene based oil-in-water emulsion system (SE) or a cationic liposome-based adjuvant significantly enhanced antibody responses against the spike receptor binding domain (RBD). Kinetics of antibody responses differed, with SE providing the most rapid response. Neutralizing antibodies developed after a single immunization in all adjuvanted groups with ID50 titers ranging from 86–4063. Spike-specific CD4 T helper responses were also elicited, comprising mainly of IFN-γ and IL-17 producing cells in the cationic liposome adjuvanted group, and more IL-5- and IL-10-secreting cells in the AH group. Interpretation: These results demonstrate that adjuvanted spike protein subunit vaccine is a viable strategy for rapidly eliciting SARS-CoV-2 neutralizing antibodies and CD4 T cell responses of various qualities depending on the adjuvant used, which can be explored in further vaccine development against COVID-19. Funding: This work was supported by the European Union Horizon 2020 research and innovation program under grant agreement no. 101003653.
Keywords	SARS-CoV-2 ; Alum ; Subunit vaccine ; CAF®01 ; MF59TM ; Squalene emulsion ; Medicine ; R ; Medicine (General) ; R5-920
Subject code	570
Language	English
Publishing date	2021-01-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Adjuvanted SARS-CoV-2 Spike Protein Elicits Neutralizing Antibodies and CD4 T Cell Responses after a Single Immunization in Mice

Wørzner, Katharina / Sheward, Daniel J. / Schmidt, Signe Tandrup / Hanke, Leo / Zimmermann, Julie / McInerney, Gerald / Karlsson Hedestam, Gunilla B. / Murrel, Ben / Christensen, Dennis / Pedersen, Gabriel

SSRN Electronic Journal ; ISSN 1556-5068

2020

Keywords	covid19
Language	English
Publisher	Elsevier BV
Publishing country	us
Document type	Article ; Online
DOI	10.2139/ssrn.3701554
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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