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Article ; Online: Glioblastoma Immune Landscape and the Potential of New Immunotherapies.

Daubon, Thomas / Hemadou, Audrey / Romero Garmendia, Irati / Saleh, Maya

2020 Volume 11, Page(s) 585616

Abstract: Glioblastoma (GBM) are the most common tumors of the central nervous system and among the deadliest cancers in adults. GBM overall survival has not improved over the last decade despite optimization of therapeutic standard-of-care. While immune ... ...

Abstract	Glioblastoma (GBM) are the most common tumors of the central nervous system and among the deadliest cancers in adults. GBM overall survival has not improved over the last decade despite optimization of therapeutic standard-of-care. While immune checkpoint inhibitors (ICI) have revolutionized cancer care, they unfortunately have little therapeutic success in GBM. Here, we elaborate on normal brain and GBM-associated immune landscapes. We describe the role of microglia and tumor-associated macrophages (TAMs) in immune suppression and highlight the impact of energy metabolism in immune evasion. We also describe the challenges and opportunities of immunotherapies in GBM and discuss new avenues based on harnessing the anti-tumor activity of myeloid cells, vaccines, chimeric antigen receptors (CAR)-T and -NK cells, oncolytic viruses, nanocarriers, and combination therapies.
MeSH term(s)	Animals ; Brain Neoplasms/immunology ; Brain Neoplasms/therapy ; Glioblastoma/immunology ; Glioblastoma/therapy ; Humans ; Immunotherapy/methods ; Tumor Escape/immunology
Language	English
Publishing date	2020-10-14
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2020.585616
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Article ; Online: A Nano-Emulsion Platform Functionalized with a Fully Human scFv-Fc Antibody for Atheroma Targeting: Towards a Theranostic Approach to Atherosclerosis.

Bonnet, Samuel / Prévot, Geoffrey / Mornet, Stéphane / Jacobin-Valat, Marie-Josée / Mousli, Yannick / Hemadou, Audrey / Duttine, Mathieu / Trotier, Aurélien / Sanchez, Stéphane / Duonor-Cérutti, Martine / Crauste-Manciet, Sylvie / Clofent-Sanchez, Gisèle

International journal of molecular sciences

2021 Volume 22, Issue 10

Abstract: Atherosclerosis is at the onset of the cardiovascular diseases that are among the leading causes of death worldwide. Currently, high-risk plaques, also called vulnerable atheromatous plaques, remain often undiagnosed until the occurrence of severe ... ...

Abstract	Atherosclerosis is at the onset of the cardiovascular diseases that are among the leading causes of death worldwide. Currently, high-risk plaques, also called vulnerable atheromatous plaques, remain often undiagnosed until the occurrence of severe complications, such as stroke or myocardial infarction. Molecular imaging agents that target high-risk atheromatous lesions could greatly improve the diagnosis of atherosclerosis by identifying sites of high disease activity. Moreover, a "theranostic approach" that combines molecular imaging agents (for diagnosis) and therapeutic molecules would be of great value for the local management of atheromatous plaques. The aim of this study was to develop and characterize an innovative theranostic tool for atherosclerosis. We engineered oil-in-water nano-emulsions (NEs) loaded with superparamagnetic iron oxide (SPIO) nanoparticles for magnetic resonance imaging (MRI) purposes. Dynamic MRI showed that NE-SPIO nanoparticles decorated with a polyethylene glycol (PEG) layer reduced their liver uptake and extended their half-life. Next, the NE-SPIO-PEG formulation was functionalized with a fully human scFv-Fc antibody (P3) recognizing galectin 3, an atherosclerosis biomarker. The P3-functionalized formulation targeted atheromatous plaques, as demonstrated in an immunohistochemistry analyses of mouse aorta and human artery sections and in an
MeSH term(s)	Animals ; Atherosclerosis/immunology ; Atherosclerosis/pathology ; Contrast Media ; Emulsions ; Female ; Humans ; Magnetic Resonance Imaging ; Magnetite Nanoparticles/administration & dosage ; Magnetite Nanoparticles/chemistry ; Mice ; Mice, Inbred C57BL ; Mice, Knockout, ApoE ; Molecular Imaging/methods ; Polyethylene Glycols ; Single-Chain Antibodies/immunology ; Theranostic Nanomedicine/methods
Chemical Substances	Contrast Media ; Emulsions ; Magnetite Nanoparticles ; Single-Chain Antibodies ; Polyethylene Glycols (3WJQ0SDW1A)
Language	English
Publishing date	2021-05-14
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms22105188
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Article ; Online: In Vivo Human Single-Chain Fragment Variable Phage Display-Assisted Identification of Galectin-3 as a New Biomarker of Atherosclerosis.

Hemadou, Audrey / Fontayne, Alexandre / Laroche-Traineau, Jeanny / Ottones, Florence / Mondon, Philippe / Claverol, Stéphane / Ducasse, Éric / Sanchez, Stéphane / Mohamad, Sarah / Lorenzato, Cyril / Duonor-Cerutti, Martine / Clofent-Sanchez, Gisèle / Jacobin-Valat, Marie-Josée

Journal of the American Heart Association

2021 Volume 10, Issue 19, Page(s) e016287

Abstract: Background Atherosclerosis is a complex pathology in which dysfunctional endothelium, activated leucocytes, macrophages, and lipid-laden foam cells are implicated, and in which plaque disruption is driven by many putative actors. This study aimed to ... ...

Abstract	Background Atherosclerosis is a complex pathology in which dysfunctional endothelium, activated leucocytes, macrophages, and lipid-laden foam cells are implicated, and in which plaque disruption is driven by many putative actors. This study aimed to identify accurate targetable biomarkers using new in vivo approaches to propose tools for improved diagnosis and treatment. Methods and Results Human scFv (single-chain fragment variable) selected by in vivo phage display in a rabbit model of atherosclerosis was reformatted as scFv fused to the scFv-Fc (single-chain fragment variable fused to the crystallizable fragment of immunoglobulin G format) antibodies. Their reactivity was tested using flow cytometry and immunoassays, and aorta sections from animal models and human carotid and coronary artery specimens. A pool of atherosclerotic proteins from human endarterectomies was co-immunoprecipitated with the selected scFv-Fc followed by mass spectrometry for target identification. Near-infrared fluorescence imaging was performed in
MeSH term(s)	Animals ; Apolipoproteins E ; Atherosclerosis/diagnosis ; Atherosclerosis/genetics ; Bacteriophages ; Biomarkers ; Galectin 3/genetics ; Humans ; Mice ; Plaque, Atherosclerotic ; Rabbits ; Single-Chain Antibodies/genetics
Chemical Substances	Apolipoproteins E ; Biomarkers ; Galectin 3 ; Single-Chain Antibodies
Language	English
Publishing date	2021-09-25
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2653953-6
ISSN	2047-9980 ; 2047-9980
ISSN (online)	2047-9980
ISSN	2047-9980
DOI	10.1161/JAHA.120.016287
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Article ; Online: Development of anti-chloro

Lokeshwaran, Kirubanandhan / Hemadou, Audrey / Jayaprakash, N S / Prasanna, R R / Jacobin-Valat, Marie-Josée / Dieryck, Wilfrid / Joucla, Gilles / Vijayalakshmi, M A / Clofent-Sanchez, Gisèle / Santarelli, Xavier / Venkataraman, Krishnan

Journal of immunological methods

2019 Volume 474, Page(s) 112637

Abstract: High density lipoproteins (HDL) are considered cardio protective. Apolipoprotein A-I (apoA-I), a major component of HDL helps in reverse cholesterol transport, whose function is greatly affected during atherosclerosis due to oxidation by myeloperoxidase. ...

Abstract	High density lipoproteins (HDL) are considered cardio protective. Apolipoprotein A-I (apoA-I), a major component of HDL helps in reverse cholesterol transport, whose function is greatly affected during atherosclerosis due to oxidation by myeloperoxidase. Amino acid tyrosine residue of apoA-I at position 192 and 166 are sensitive to oxidation by myeloperoxidase resulting in the generation of chlorinated and nitrated apoA-I and they are believed to be present in atherosclerotic plaques and in circulation. These oxidized apoA-I have been suggested as potential indicator(s) of CVD risks in humans. To detect the levels of oxidized apoA-I there is a need for developing monoclonal antibodies (mAbs) with high specificity and sensitivity that could be utilized routinely in clinical immune based assays for blood plasma or for in vivo imaging. In this study, chemically chlorinated apoA-I (chlorinated
MeSH term(s)	Animals ; Antibodies, Monoclonal/immunology ; Antibody Specificity ; Apolipoprotein A-I/analysis ; Apolipoprotein A-I/immunology ; Atherosclerosis/diagnosis ; Atherosclerosis/genetics ; Atherosclerosis/immunology ; Atherosclerosis/metabolism ; Biomarkers/analysis ; Coronary Artery Disease/diagnosis ; Coronary Artery Disease/immunology ; Coronary Artery Disease/metabolism ; Disease Models, Animal ; Halogenation ; Humans ; Immunologic Tests ; Lipoproteins, HDL/analysis ; Lipoproteins, HDL/immunology ; Mice, Knockout, ApoE ; Oxidation-Reduction ; Plaque, Atherosclerotic ; Predictive Value of Tests ; Reproducibility of Results ; Tyrosine
Chemical Substances	APOA1 protein, human ; Antibodies, Monoclonal ; Apolipoprotein A-I ; Biomarkers ; Lipoproteins, HDL ; Tyrosine (42HK56048U)
Language	English
Publishing date	2019-08-03
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	120142-6
ISSN	1872-7905 ; 0022-1759
ISSN (online)	1872-7905
ISSN	0022-1759
DOI	10.1016/j.jim.2019.112637
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Article: Development of anti-chloro 192 tyrosine HDL apoA-I antibodies for the immunodiagnosis of cardiovascular diseases

Lokeshwaran, Kirubanandhan / Hemadou, Audrey / Jayaprakash, N.S / Prasanna, R.R / Jacobin-Valat, Marie-Josée / Dieryck, Wilfrid / Joucla, Gilles / Vijayalakshmi, M.A / Clofent-Sanchez, Gisèle / Santarelli, Xavier / Venkataraman, Krishnan

Journal of immunological methods. 2019 July 26,

2019

Abstract	High density lipoproteins (HDL) are considered cardio protective. Apolipoprotein A-I (apoA-I), a major component of HDL helps in reverse cholesterol transport, whose function is greatly affected during atherosclerosis due to oxidation by myeloperoxidase. Amino acid tyrosine residue of apoA-I at position 192 and 166 are sensitive to oxidation by myeloperoxidase resulting in the generation of chlorinated and nitrated apoA-I and they are believed to be present in atherosclerotic plaques and in circulation. These oxidized apoA-I have been suggested as potential indicator(s) of CVD risks in humans. To detect the levels of oxidized apoA-I there is a need for developing monoclonal antibodies (mAbs) with high specificity and sensitivity that could be utilized routinely in clinical immune based assays for blood plasma or for in vivo imaging. In this study, chemically chlorinated apoA-I (chlorinated 192tyrosine- apoA-I) and a short synthetic peptide, containing the corresponding chlorinated tyrosine residue, conjugated to keyhole limpet hemocyanin (KLH) carrier protein were used for immunization. Stable hybridoma clones F7D5 and G11E3 were found to be highly sensitive and reactive towards chlorinated 192tyrosine- apoA-I. Interestingly, these mAbs also displayed positive reaction with atherosclerotic plaques obtained from mouse and human biopsies. In vitro or in vivo diagnostic tests could be developed either by detecting oxidized apoA-I in human plasma or by directly imaging atheroma plaques as both mAbs were shown to stain human atheroma. The anti-chlorinated 192tyrosine- apoA-I mAbs described in this study may have a high diagnostic potential in predicting CVD risks.
Keywords	apolipoprotein A-I ; atherosclerosis ; biopsy ; blood plasma ; cholesterol ; clones ; diagnostic techniques ; high density lipoprotein ; humans ; hybridomas ; image analysis ; immunization ; keyhole limpet hemocyanin ; mice ; monoclonal antibodies ; myeloperoxidase ; oxidation ; prediction ; risk ; serodiagnosis ; synthetic peptides ; tyrosine
Language	English
Dates of publication	2019-0726
Publishing place	Elsevier B.V.
Document type	Article
Note	Pre-press version
ZDB-ID	120142-6
ISSN	1872-7905 ; 0022-1759
ISSN (online)	1872-7905
ISSN	0022-1759
DOI	10.1016/j.jim.2019.112637
Database	NAL-Catalogue (AGRICOLA)

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Zs.A 795: Show issues

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Article ; Online: An innovative flow cytometry method to screen human scFv-phages selected by in vivo phage-display in an animal model of atherosclerosis.

Hemadou, Audrey / Laroche-Traineau, Jeanny / Antoine, Ségolène / Mondon, Philippe / Fontayne, Alexandre / Le Priol, Yannick / Claverol, Stéphane / Sanchez, Stéphane / Cerutti, Martine / Ottones, Florence / Clofent-Sanchez, Gisèle / Jacobin-Valat, Marie-Josée

Scientific reports

2018 Volume 8, Issue 1, Page(s) 15016

Abstract: Atherosclerosis is a chronic, progressive inflammatory disease that may develop into vulnerable lesions leading to thrombosis. This pathology is characterized by the deposition of lipids within the arterial wall and infiltration of immune cells leading ... ...

Abstract	Atherosclerosis is a chronic, progressive inflammatory disease that may develop into vulnerable lesions leading to thrombosis. This pathology is characterized by the deposition of lipids within the arterial wall and infiltration of immune cells leading to amplification of inflammation. Nowadays there is a rising interest to assess directly the molecular and cellular components that underlie the clinical condition of stroke and myocardial infarction. Single chain fragment variable (scFv)-phages issuing from a human combinatorial library were selected on the lesions induced in a rabbit model of atherosclerosis after three rounds of in vivo phage display. We further implemented a high-throughput flow cytometry method on rabbit protein extracts to individually test one thousand of scFv-phages. Two hundred and nine clones were retrieved on the basis of their specificity for atherosclerotic proteins. Immunohistochemistry assays confirmed the robustness of the designed cytometry protocol. Sequencing of candidates demonstrated their high diversity in VH and VL germline usage. The large number of candidates and their diversity open the way in the discovery of new biomarkers. Here, we successfully showed the capacity of combining in vivo phage display and high-throughput cytometry strategies to give new insights in in vivo targetable up-regulated biomarkers in atherosclerosis.
MeSH term(s)	Animals ; Antibodies, Monoclonal/genetics ; Antibodies, Monoclonal/immunology ; Atherosclerosis/genetics ; Atherosclerosis/immunology ; Atherosclerosis/pathology ; Cell Surface Display Techniques ; Disease Models, Animal ; Flow Cytometry ; Humans ; Immunohistochemistry/methods ; Rabbits ; Single-Chain Antibodies/immunology ; Single-Chain Antibodies/isolation & purification
Chemical Substances	Antibodies, Monoclonal ; Single-Chain Antibodies
Language	English
Publishing date	2018-10-09
Publishing country	England
Document type	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/s41598-018-33382-2
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Article ; Online: Multimodal molecular imaging of atherosclerosis: Nanoparticles functionalized with scFv fragments of an anti-αIIbβ3 antibody.

Larivière, Mélusine / Lorenzato, Cyril Samuel / Adumeau, Laurent / Bonnet, Samuel / Hémadou, Audrey / Jacobin-Valat, Marie-Josée / Noubhani, Abdelmajid / Santarelli, Xavier / Minder, Laetitia / Di Primo, Carmelo / Sanchez, Stéphane / Mornet, Stéphane / Laroche-Traineau, Jeanny / Clofent-Sanchez, Gisèle

Nanomedicine : nanotechnology, biology, and medicine

2019 Volume 22, Page(s) 102082

Abstract: Due to the wealth of actors involved in the development of atherosclerosis, molecular imaging based on the targeting of specific markers would substantiate the diagnosis of life-threatening atheroma plaques. To this end, TEG4 antibody is a promising ... ...

Abstract	Due to the wealth of actors involved in the development of atherosclerosis, molecular imaging based on the targeting of specific markers would substantiate the diagnosis of life-threatening atheroma plaques. To this end, TEG4 antibody is a promising candidate targeting the activated platelets (integrin αIIbβ3) highly represented within the plaque. In this study, scFv antibody fragments were used to functionalize multimodal imaging nanoparticles. This grafting was performed in a regio-selective way to preserve TEG4 activity and the avidity of the nanoparticles was studied with respect to the number of grafted antibodies. Subsequently, taking advantage of the nanoparticle bimodality, both near infrared fluorescence and magnetic resonance imaging of the atheroma plaque were performed in the ApoE
MeSH term(s)	Animals ; Atherosclerosis/diagnosis ; Atherosclerosis/pathology ; Fluorescence ; Magnetic Resonance Imaging ; Male ; Mice ; Molecular Imaging ; Multimodal Imaging ; Nanoparticles/chemistry ; Platelet Glycoprotein GPIIb-IIIa Complex/immunology ; Rabbits ; Single-Chain Antibodies/immunology ; Tissue Distribution
Chemical Substances	Platelet Glycoprotein GPIIb-IIIa Complex ; Single-Chain Antibodies
Language	English
Publishing date	2019-08-09
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2183417-9
ISSN	1549-9642 ; 1549-9634
ISSN (online)	1549-9642
ISSN	1549-9634
DOI	10.1016/j.nano.2019.102082
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Article ; Online: A Recombinant Human Anti-Platelet scFv Antibody Produced in Pichia pastoris for Atheroma Targeting.

Vallet-Courbin, Amelie / Larivière, Mélusine / Hocquellet, Agnès / Hemadou, Audrey / Parimala, Sarjapura-Nagaraja / Laroche-Traineau, Jeanny / Santarelli, Xavier / Clofent-Sanchez, Gisèle / Jacobin-Valat, Marie-Josée / Noubhani, Abdelmajid

PloS one

2017 Volume 12, Issue 1, Page(s) e0170305

Abstract: Cells of the innate and adaptive immune system are key factors in the progression of atherosclerotic plaque, leading to plaque instability and rupture, potentially resulting in acute atherothrombotic events such as coronary artery disease, ... ...

Abstract	Cells of the innate and adaptive immune system are key factors in the progression of atherosclerotic plaque, leading to plaque instability and rupture, potentially resulting in acute atherothrombotic events such as coronary artery disease, cerebrovascular disease and peripheral arterial disease. Here, we describe the cloning, expression, purification, and immunoreactivity assessment of a recombinant single-chain variable fragment (scFv) derived from a human anti-αIIbβ3 antibody (HuAb) selected to target atheromatous lesions for the presence of platelets. Indeed, platelets within atheroma plaques have been shown to play a role in inflammation, in platelet-leucocyte aggregates and in thrombi formation and might thus be considered relevant biomarkers of atherosclerotic progression. The DNA sequence that encodes the anti-αIIbβ3 TEG4 scFv previously obtained from a phage-display selection on activated platelets, was inserted into the eukaryote vector (pPICZαA) in fusion with a tag sequence encoding 2 cysteines useable for specific probes grafting experiments. The recombinant protein was expressed at high yields in Pichia pastoris (30 mg/L culture). The advantage of P. pastoris as an expression system is the production and secretion of recombinant proteins in the supernatant, ruling out the difficulties encountered when scFv are produced in the cytoplasm of bacteria (low yield, low solubility and reduced affinity). The improved conditions allowed for the recovery of highly purified and biologically active scFv fragments ready to be grafted in a site-directed way to nanoparticles for the imaging of atherosclerotic plaques involving inflammatory processes and thus at high risk of instability.
MeSH term(s)	Blood Platelets/immunology ; Cell Surface Display Techniques ; Cloning, Molecular ; Gene Expression ; Genetic Vectors ; Humans ; Inflammation/drug therapy ; Inflammation/immunology ; Pichia/genetics ; Plaque, Atherosclerotic/drug therapy ; Plaque, Atherosclerotic/immunology ; Platelet Aggregation/immunology ; Recombinant Proteins/immunology ; Recombinant Proteins/therapeutic use ; Single-Chain Antibodies/immunology ; Single-Chain Antibodies/therapeutic use
Chemical Substances	Recombinant Proteins ; Single-Chain Antibodies
Language	English
Publishing date	2017
Publishing country	United States
Document type	Journal Article
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0170305
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Article ; Online: Solid Lipid Nanoparticles for Image-Guided Therapy of Atherosclerosis.

Oumzil, Khalid / Ramin, Michael A / Lorenzato, Cyril / Hémadou, Audrey / Laroche, Jeanny / Jacobin-Valat, Marie Josée / Mornet, Stephane / Roy, Claude-Eric / Kauss, Tina / Gaudin, Karen / Clofent-Sanchez, Gisèle / Barthélémy, Philippe

Bioconjugate chemistry

2016 Volume 27, Issue 3, Page(s) 569–575

Abstract: Although the application of nanotechnologies to atherosclerosis remains a young field, novel strategies are needed to address this public health issue. In this context, the magnetic resonance imaging (MRI) approach has been gradually investigated in ... ...

Abstract	Although the application of nanotechnologies to atherosclerosis remains a young field, novel strategies are needed to address this public health issue. In this context, the magnetic resonance imaging (MRI) approach has been gradually investigated in order to enable image-guided treatments. In this contribution, we report a new approach based on nucleoside-lipids allowing the synthesis of solid lipid nanoparticles (SLN) loaded with iron oxide particles and therapeutic agents. The insertion of nucleoside-lipids allows the formation of stable SLNs loaded with prostacycline (PGI2) able to inhibit platelet aggregation. The new SLNs feature better relaxivity properties in comparison to the clinically used contrast agent Feridex, indicating that SLNs are suitable for image-guided therapy.
MeSH term(s)	Atherosclerosis/therapy ; Epoprostenol/administration & dosage ; Epoprostenol/therapeutic use ; Lipids/chemistry ; Magnetic Resonance Imaging/methods ; Nanoparticles
Chemical Substances	Lipids ; Epoprostenol (DCR9Z582X0)
Language	English
Publishing date	2016-03-16
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1024041-x
ISSN	1520-4812 ; 1043-1802
ISSN (online)	1520-4812
ISSN	1043-1802
DOI	10.1021/acs.bioconjchem.5b00590
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Article: Pacific Biosciences Sequencing and IMGT/HighV-QUEST Analysis of Full-Length Single Chain Fragment Variable from an

Hemadou, Audrey / Giudicelli, Véronique / Smith, Melissa Laird / Lefranc, Marie-Paule / Duroux, Patrice / Kossida, Sofia / Heiner, Cheryl / Hepler, N Lance / Kuijpers, John / Groppi, Alexis / Korlach, Jonas / Mondon, Philippe / Ottones, Florence / Jacobin-Valat, Marie-Josée / Laroche-Traineau, Jeanny / Clofent-Sanchez, Gisèle

Frontiers in immunology

2017 Volume 8, Page(s) 1796

Abstract: Phage-display selection of immunoglobulin (IG) or antibody single chain Fragment variable (scFv) from combinatorial libraries is widely used for identifying new antibodies for novel targets. Next-generation sequencing (NGS) has recently emerged as a new ... ...

Abstract	Phage-display selection of immunoglobulin (IG) or antibody single chain Fragment variable (scFv) from combinatorial libraries is widely used for identifying new antibodies for novel targets. Next-generation sequencing (NGS) has recently emerged as a new method for the high throughput characterization of IG and T cell receptor (TR) immune repertoires both
Language	English
Publishing date	2017
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2606827-8
ISSN	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2017.01796
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