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Article ; Online: High performance production process development and scale-up of an anti-TSLP nanobody.

Li, Xiaofei / Qiao, Peng / Zhang, Yicai / Liu, Guoxin / Zhu, Min / Gai, Junwei / Wan, Yakun

2024 Volume 218, Page(s) 106441

Abstract: Nanobodies (Nbs) represent a class of single-domain antibodies with great potential application value across diverse biotechnology fields, including therapy and diagnostics. Thymic Stromal Lymphopoietin (TSLP) is an epithelial cell-derived cytokine, ... ...

Abstract	Nanobodies (Nbs) represent a class of single-domain antibodies with great potential application value across diverse biotechnology fields, including therapy and diagnostics. Thymic Stromal Lymphopoietin (TSLP) is an epithelial cell-derived cytokine, playing a crucial role in the regulation of type 2 immune responses at barrier surfaces such as skin and the respiratory/gastrointestinal tract. In this study, a method for the expression and purification of anti-TSLP nanobody (Nb3341) was established at 7 L scale and subsequently scaled up to 100 L scale. Key parameters, including induction temperature, methanol feed and induction pH were identified as key factors by Plackett-Burman design (PBD) and were optimized in 7 L bioreactor, yielding optimal values of 24 °C, 8.5 mL/L/h and 6.5, respectively. Furthermore, Diamond Mix-A and Diamond MMC were demonstrated to be the optimal capture and polishing resins. The expression and purification process of Nb3341 at 100L scale resulted in 22.97 g/L titer, 98.7% SEC-HPLC purity, 95.7% AEX-HPLC purity, 4 ppm of HCP content and 1 pg/mg of HCD residue. The parameters of the scaling-up process were consistent with the results of the optimized process, further demonstrating the feasibility and stability of this method. This study provides a highly promising and competitive approach for transitioning from laboratory-scale to commercial production-scale of nanobodies.
MeSH term(s)	Thymic Stromal Lymphopoietin ; Single-Domain Antibodies/genetics ; Single-Domain Antibodies/metabolism ; Cytokines/metabolism ; Epithelial Cells ; Diamond/metabolism
Chemical Substances	Thymic Stromal Lymphopoietin (GT0IL38SP4) ; Single-Domain Antibodies ; Cytokines ; Diamond (7782-40-3)
Language	English
Publishing date	2024-02-16
Publishing country	United States
Document type	Journal Article
ZDB-ID	1055455-5
ISSN	1096-0279 ; 1046-5928
ISSN (online)	1096-0279
ISSN	1046-5928
DOI	10.1016/j.pep.2024.106441
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Article ; Online: Identification and Characterization of a Novel Nanobody Against Human CTGF to Reveal Its Antifibrotic Effect in an in vitro Model of Liver Fibrosis.

Liu, Rong / Zhu, Min / Chen, Jiaojiao / Gai, Junwei / Huang, Jing / Zhou, Yingqun / Wan, Yakun / Tu, Chuantao

International journal of nanomedicine

2023 Volume 18, Page(s) 5407–5422

Abstract: Background: No agents are currently available for the treatment or reversal of liver fibrosis. Novel antifibrotic therapies for chronic liver diseases are thus urgently needed. Connective tissue growth factor (CTGF) has been shown to contributes ... ...

Abstract	Background: No agents are currently available for the treatment or reversal of liver fibrosis. Novel antifibrotic therapies for chronic liver diseases are thus urgently needed. Connective tissue growth factor (CTGF) has been shown to contributes profoundly to liver fibrogenesis, which makes CTGF as a promising target for developing antifibrotic agents. Methods: In this study, we identified a novel nanobody (Nb) against human CTGF (anti-CTGF Nb) by phage display using an immunized camel, which showed high affinity and specificity in vitro. LX-2 cells, the immortalized human hepatic stellate cells, were induced by transforming growth factor beta1 (TGFβ1) as an in vitro model of liver fibrosis to verify the antifibrotic activity of the anti-CTGF Nb. Results: Our data demonstrated that anti-CTGF Nb effectively alleviated TGFβ1-induced LX-2 cell proliferation, activation, and migration, and promoted the apoptosis of activated LX-2 cells in response to TGFβ1. Moreover, the anti-CTGF Nb remarkably reduced the levels of TGFβ1, Smad2, and Smad3 expression in LX-2 stellate cells stimulated by TGFβ1. Conclusion: Taken together, we successfully identified a novel Nb against human CTGF, which exhibited antifibrotic effects in vitro by regulating the biological functions of human stellate cells LX-2.
Language	English
Publishing date	2023-09-21
Publishing country	New Zealand
Document type	Journal Article
ZDB-ID	2364941-0
ISSN	1178-2013 ; 1176-9114
ISSN (online)	1178-2013
ISSN	1176-9114
DOI	10.2147/IJN.S428430
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Article ; Online: Theoretical investigations of weakly- and strongly-coupled multi-core fibers for the applications of optical submarine communications under power and fiber count limits.

Sun, Lin / Zhang, Junwei / Zhou, Gai / Chen, Bin / Liu, Gordon Ning / Cai, Yi / Li, Zhaohui / Lu, Chao / Shen, Gangxiang

Optics express

2023 Volume 31, Issue 3, Page(s) 4615–4629

Abstract: The practical cable design for optical submarine communications has a limited fiber pair count due to the mechanical considerations of cable weight and size. Consequently, multi-core fibers (MCFs) could exhibit higher capacity than conventional single- ... ...

Abstract	The practical cable design for optical submarine communications has a limited fiber pair count due to the mechanical considerations of cable weight and size. Consequently, multi-core fibers (MCFs) could exhibit higher capacity than conventional single-mode fibers (SMFs) thanks to space division multiplexing (SDM). That is because the power supply to a submarine cable is fed by the voltage difference between shores. Under the power-limited condition, SDM improves the cable capacity by using more paths which outperforms the SMF link whose capacity approximately complies with a logarithmic relationship to optical power. At the same time, fiber nonlinearity can be alleviated by the reduced power density of transmitted light in MCFs, due to the increased spatial diversity and mode coupling among coupled cores. In this work, we theoretically investigate the potentials of MCFs including weakly-coupled multicore fiber (WC-MCF) and strongly-coupled multicore fiber (SC-MCF) as the propagation media for submarine communications across the Atlantic and the Pacific. To fairly compare the performances of SMFs- and MCFs-based submarine cables, the Gaussian noise (GN) model for SDM links is employed to optimize the systematic settings including spatial multiplicity and single span length. Then, we develop an SDM and wavelength division multiplexing (WDM) fiber transmission model based on coupled nonlinear Schrodinger equations (CNSE) to investigate the optical filed coupling effect in MCFs-based cables. The developed transmission model has been self-examined by measuring the inter-core crosstalk (IC-XT) and spatial mode dispersion (SMD), referring to the set values. As indicated by the theoretical analysis, the WC-MCFs cable exhibits a larger capacity than the SMFs cable, when the fiber pair count is limited below 32. Moreover, the SC-MCFs cable outperforms the WC-MCFs cable thanks to the reduced fiber nonlinearity due to the random mode coupling and the assistance of multiple-input and multiple-output digital signal processing (MIMO-DSP). At last, the marginal influences of IC-XT, SMD, and insertion loss of Fan-in and Fan-out couplers are also analyzed for the MCFs cable.
Language	English
Publishing date	2023-01-30
Publishing country	United States
Document type	Journal Article
ZDB-ID	1491859-6
ISSN	1094-4087 ; 1094-4087
ISSN (online)	1094-4087
ISSN	1094-4087
DOI	10.1364/OE.480344
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Article ; Online: TROP2-directed nanobody-drug conjugate elicited potent antitumor effect in pancreatic cancer.

Xu, Caili / Zhu, Min / Wang, Qian / Cui, Jiajun / Huang, Yuping / Huang, Xiting / Huang, Jing / Gai, Junwei / Li, Guanghui / Qiao, Peng / Zeng, Xian / Ju, Dianwen / Wan, Yakun / Zhang, Xuyao

Journal of nanobiotechnology

2023 Volume 21, Issue 1, Page(s) 410

Abstract: Background: Pancreatic cancer is a highly aggressive malignancy with limited treatment options and a poor prognosis. Trophoblast cell surface antigen 2 (TROP2), a cell surface antigen overexpressed in the tumors of more than half of pancreatic cancer ... ...

Abstract	Background: Pancreatic cancer is a highly aggressive malignancy with limited treatment options and a poor prognosis. Trophoblast cell surface antigen 2 (TROP2), a cell surface antigen overexpressed in the tumors of more than half of pancreatic cancer patients, has been identified as a potential target for antibody-drug conjugates (ADCs). Almost all reported TROP2-targeted ADCs are of the IgG type and have been poorly studied in pancreatic cancer. Here, we aimed to develop a novel nanobody-drug conjugate (NDC) targeting TROP2 for the treatment of pancreatic cancer. Results: In this study, we developed a novel TROP2-targeted NDC, HuNb Conclusion: HuNb
MeSH term(s)	Humans ; Antigens, Surface ; Cell Line, Tumor ; Immunoconjugates/chemistry ; Pancreatic Neoplasms/pathology ; Xenograft Model Antitumor Assays ; Animals ; Pancreatic Neoplasms
Chemical Substances	Antigens, Surface ; Immunoconjugates ; TACSTD2 protein, human
Language	English
Publishing date	2023-11-06
Publishing country	England
Document type	Journal Article
ZDB-ID	2100022-0
ISSN	1477-3155 ; 1477-3155
ISSN (online)	1477-3155
ISSN	1477-3155
DOI	10.1186/s12951-023-02183-9
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Article: Increased H

Wahafu, Wasilijiang / Gai, Junwei / Song, Liming / Ping, Hao / Wang, Mingshuai / Yang, Feiya / Niu, Yinong / Xing, Nianzeng

Oncology letters

2018 Volume 15, Issue 6, Page(s) 8484–8490

Abstract: ... ...

Abstract	H
Language	English
Publishing date	2018-03-29
Publishing country	Greece
Document type	Journal Article
ZDB-ID	2573196-8
ISSN	1792-1082 ; 1792-1074
ISSN (online)	1792-1082
ISSN	1792-1074
DOI	10.3892/ol.2018.8373
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Article ; Online: Preclinical development of a long-acting trivalent bispecific nanobody targeting IL-5 for the treatment of eosinophilic asthma.

Ma, Linlin / Zhu, Min / Li, Guanghui / Gai, Junwei / Li, Yanfei / Gu, Huaiyu / Qiao, Peng / Li, Xiaofei / Ji, Weiwei / Zhao, Rui / Wu, Yue / Wan, Yakun

Respiratory research

2022 Volume 23, Issue 1, Page(s) 316

Abstract: Background: Eosinophilic asthma is a common subtype of severe asthma with high morbidity and mortality. The cytokine IL-5 has been shown to be a key driver of the development and progression of disease. Although approved monoclonal antibodies (mAbs) ... ...

Abstract	Background: Eosinophilic asthma is a common subtype of severe asthma with high morbidity and mortality. The cytokine IL-5 has been shown to be a key driver of the development and progression of disease. Although approved monoclonal antibodies (mAbs) targeting IL-5/IL-5R have shown good safety and efficacy, some patients have inadequate responses and frequent dosing results in medication nonadherence. Results: We constructed a novel trivalent bispecific nanobody (Nb) consisting of 3 VHHs that bind to 2 different epitopes of IL-5 and 1 epitope of albumin derived from immunized phage display libraries. This trivalent IL-5-HSA Nb exhibited similar IL-5/IL-5R blocking activities to mepolizumab (Nucala), an approved targeting IL-5 mAb. Surprisingly, this trivalent Nb was 58 times more active than mepolizumab in inhibiting TF-1-cell proliferation. In primate studies, the trivalent IL-5-HSA Nb showed excellent pharmacokinetic properties, and peripheral blood eosinophil levels remained significantly suppressed for two months after a single dose. In addition, the trivalent IL-5-HSA Nb could be produced on a large scale in a P. pastoris X-33 yeast system with high purity and good thermal stability. Conclusions: These findings suggest that the trivalent bispecific IL-5-HSA Nb has the potential to be a next-generation therapeutic agent targeting IL-5 for the treatment of severe eosinophilic asthma.
MeSH term(s)	Animals ; Interleukin-5/metabolism ; Interleukin-5/therapeutic use ; Pulmonary Eosinophilia/drug therapy ; Pulmonary Eosinophilia/metabolism ; Asthma/metabolism ; Eosinophils/metabolism ; Antibodies, Monoclonal/therapeutic use
Chemical Substances	Interleukin-5 ; Antibodies, Monoclonal
Language	English
Publishing date	2022-11-19
Publishing country	England
Document type	Journal Article
ZDB-ID	2041675-1
ISSN	1465-993X ; 1465-993X
ISSN (online)	1465-993X
ISSN	1465-993X
DOI	10.1186/s12931-022-02240-1
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Article ; Online: Preclinical development of a novel CD47 nanobody with less toxicity and enhanced anti-cancer therapeutic potential.

Ma, Linlin / Zhu, Min / Gai, Junwei / Li, Guanghui / Chang, Qing / Qiao, Peng / Cao, Longlong / Chen, Wanqing / Zhang, Siyuan / Wan, Yakun

Journal of nanobiotechnology

2020 Volume 18, Issue 1, Page(s) 12

Abstract: Background: CD47, the integrin-related protein, plays an important role in immune resistance and escape of tumor cells. Antibodies blocking the CD47/SIRPα signal pathway can effectively stimulate macrophage-mediated phagocytosis of tumor cells, which ... ...

Abstract	Background: CD47, the integrin-related protein, plays an important role in immune resistance and escape of tumor cells. Antibodies blocking the CD47/SIRPα signal pathway can effectively stimulate macrophage-mediated phagocytosis of tumor cells, which becomes a promising approach for tumor immunotherapy. Nanobodies (Nbs) derived from camelid animals are emerging as a new force in antibody therapy. Results: HuNb1-IgG4, an innovative anti-CD47 nanobody, was developed with high affinity and specificity. It effectively enhanced macrophage-mediated phagocytosis of tumor cells in vitro and showed potent anti-ovarian and anti-lymphoma activity in vivo. Importantly, HuNb1-IgG4 did not induce the agglutination of human red blood cells (RBCs) in vitro and exhibited high safety for hematopoietic system in cynomolgus monkey. In addition, HuNb1-IgG4 could be produced on a large scale in CHO-S cells with high activity and good stability. Also, we established anti-CD47/CD20 bispecific antibody (BsAb) consisted of HuNb1 and Rituximab, showing more preference binding to tumor cells and more potent anti-lymphoma activity compared to HuNb1-IgG4. Conclusions: Both of HuNb1-IgG4 and anti-CD47/CD20 BsAb are potent antagonists of CD47/SIRPα pathway and promising candidates for clinical trials.
MeSH term(s)	Animals ; Antineoplastic Agents/pharmacology ; CD47 Antigen/immunology ; Cell Line ; Cell Surface Display Techniques ; Female ; Humans ; Immunoglobulin G/metabolism ; Macaca fascicularis ; Mice, Inbred NOD ; Recombinant Fusion Proteins/metabolism ; Single-Domain Antibodies/pharmacology ; Single-Domain Antibodies/toxicity
Chemical Substances	Antineoplastic Agents ; CD47 Antigen ; Immunoglobulin G ; Recombinant Fusion Proteins ; Single-Domain Antibodies
Language	English
Publishing date	2020-01-13
Publishing country	England
Document type	Journal Article
ISSN	1477-3155
ISSN (online)	1477-3155
DOI	10.1186/s12951-020-0571-2
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Article ; Online: A novel bispecific nanobody with PD-L1/TIGIT dual immune checkpoint blockade.

Ma, Linlin / Gai, Junwei / Qiao, Peng / Li, Yanfei / Li, Xiaofei / Zhu, Min / Li, Guanghui / Wan, Yakun

Biochemical and biophysical research communications

2020 Volume 531, Issue 2, Page(s) 144–151

Abstract: Cancer immunotherapy have changed the paradigm of cancer treatment, but there remains a great need for improvement given that less patients with tumors respond to the treatment of PD-1/PD-L1 blockade. TIGIT (also called T cell immunoreceptor with Ig and ... ...

Abstract	Cancer immunotherapy have changed the paradigm of cancer treatment, but there remains a great need for improvement given that less patients with tumors respond to the treatment of PD-1/PD-L1 blockade. TIGIT (also called T cell immunoreceptor with Ig and ITIM domains), a novel immune checkpoint molecule, has been shown a promising target for drug development of immunotherapy. Here we report generation and characterization of a multivalent bispecific antibody (BsAb) that co-targets PD-L1 and TIGIT. The BsAb consists of tetravalent anti-PD-L1 Fc-fusion nanobody (Nb) and tetravalent anti-TIGIT Nb. The parental anti-PD-L1 Nb showed high specificity and affinity to primate PD-L1, the enhanced T cell activity in vitro and anti-tumor activity in vivo. Similarly, the parental anti-TIGIT Nb showed the high specificity and affinity to primate TIGIT and the enhanced T cell activity. Furthermore, we demonstrated that the BsAb retained high blocking activity towards PD-1/PD-L1 or TIGIT/CD155 interaction. The BsAb synergistically enhanced T cell activities in vitro compared to two parental Nbs. Taken together, we obtained a multivalent BsAb blocking biological function of PD-L1 and TIGIT and it is worthy to further study the anti-tumor activities of this BsAb in vivo.
MeSH term(s)	Animals ; Antibodies, Bispecific/immunology ; Antibodies, Blocking/immunology ; Antibody Affinity/immunology ; B7-H1 Antigen/metabolism ; Cell Line ; Female ; Humans ; Immune Checkpoint Inhibitors/pharmacology ; Mice ; Receptors, Fc/metabolism ; Receptors, Immunologic/metabolism ; Single-Domain Antibodies/immunology ; T-Lymphocytes/immunology
Chemical Substances	Antibodies, Bispecific ; Antibodies, Blocking ; B7-H1 Antigen ; Immune Checkpoint Inhibitors ; Receptors, Fc ; Receptors, Immunologic ; Single-Domain Antibodies ; TIGIT protein, human
Language	English
Publishing date	2020-08-08
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	205723-2
ISSN	1090-2104 ; 0006-291X ; 0006-291X
ISSN (online)	1090-2104 ; 0006-291X
ISSN	0006-291X
DOI	10.1016/j.bbrc.2020.07.072
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Zs.A 116: Show issues

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Article ; Online: A potent neutralizing nanobody against SARS-CoV-2 with inhaled delivery potential.

Gai, Junwei / Ma, Linlin / Li, Guanghui / Zhu, Min / Qiao, Peng / Li, Xiaofei / Zhang, Haiwei / Zhang, Yanmin / Chen, Yadong / Ji, Weiwei / Zhang, Hao / Cao, Huanhuan / Li, Xionghui / Gong, Rui / Wan, Yakun

MedComm

2021 Volume 2, Issue 1, Page(s) 101–113

Abstract: The coronavirus disease 2019 (COVID-19) pandemic has become a serious burden on global public health. Although therapeutic drugs against COVID-19 have been used in many countries, their efficacy is still limited. We here reported nanobody (Nb) phage ... ...

Abstract	The coronavirus disease 2019 (COVID-19) pandemic has become a serious burden on global public health. Although therapeutic drugs against COVID-19 have been used in many countries, their efficacy is still limited. We here reported nanobody (Nb) phage display libraries derived from four camels immunized with the SARS-CoV-2 spike receptor-binding domain (RBD), from which 381 Nbs were identified to recognize SARS-CoV-2-RBD. Furthermore, seven Nbs were shown to block interaction of human angiotensin-converting enzyme 2 (ACE2) with SARS-CoV-2-RBD variants and two Nbs blocked the interaction of human ACE2 with bat-SL-CoV-WIV1-RBD and SARS-CoV-1-RBD. Among these candidates, Nb11-59 exhibited the highest activity against authentic SARS-CoV-2 with 50% neutralizing dose (ND
Language	English
Publishing date	2021-03-04
Publishing country	China
Document type	Journal Article
ISSN	2688-2663
ISSN (online)	2688-2663
DOI	10.1002/mco2.60
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Article: A novel bispecific nanobody with PD-L1/TIGIT dual immune checkpoint blockade

Ma, Linlin / Gai, Junwei / Qiao, Peng / Li, Yanfei / Li, Xiaofei / Zhu, Min / Li, Guanghui / Wan, Yakun

Biochemical and biophysical research communications. 2020 Oct. 15, v. 531, no. 2

2020

Abstract	Cancer immunotherapy have changed the paradigm of cancer treatment, but there remains a great need for improvement given that less patients with tumors respond to the treatment of PD-1/PD-L1 blockade. TIGIT (also called T cell immunoreceptor with Ig and ITIM domains), a novel immune checkpoint molecule, has been shown a promising target for drug development of immunotherapy. Here we report generation and characterization of a multivalent bispecific antibody (BsAb) that co-targets PD-L1 and TIGIT. The BsAb consists of tetravalent anti-PD-L1 Fc-fusion nanobody (Nb) and tetravalent anti-TIGIT Nb. The parental anti-PD-L1 Nb showed high specificity and affinity to primate PD-L1, the enhanced T cell activity in vitro and anti-tumor activity in vivo. Similarly, the parental anti-TIGIT Nb showed the high specificity and affinity to primate TIGIT and the enhanced T cell activity. Furthermore, we demonstrated that the BsAb retained high blocking activity towards PD-1/PD-L1 or TIGIT/CD155 interaction. The BsAb synergistically enhanced T cell activities in vitro compared to two parental Nbs. Taken together, we obtained a multivalent BsAb blocking biological function of PD-L1 and TIGIT and it is worthy to further study the anti-tumor activities of this BsAb in vivo.
Keywords	T-lymphocytes ; antibodies ; antineoplastic activity ; cancer therapy ; drug development ; immunologic receptors ; immunotherapy ; research
Language	English
Dates of publication	2020-1015
Size	p. 144-151.
Publishing place	Elsevier Inc.
Document type	Article
Note	NAL-AP-2-clean
ZDB-ID	205723-2
ISSN	0006-291X ; 0006-291X
ISSN (online)	0006-291X
ISSN	0006-291X
DOI	10.1016/j.bbrc.2020.07.072
Database	NAL-Catalogue (AGRICOLA)

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