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Purification of Protein-complexes from the Cyanobacterium Synechocystis sp. PCC 6803 Using FLAG-affinity Chromatography

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cDNA Display Mediated Immuno-PCR (cD-IPCR): A Novel PCR-based Antigen Detection Method

Authors: Chathuni Jayathilake, Takuya Terai and Naoto Nemoto, date: 12/20/2019, view: 1301, Q&A: 0

[Abstract] Immuno-PCR (IPCR) is a powerful method in antigen detection where a PCR-amplifiable DNA reporter is conjugated to a specific antibody or an aptamer for the target molecule. In the development and application of IPCR, successful conjugation of a protein (an antibody) with a reporter DNA becomes challenging. To address this issue, we recently ...

A Refined Protocol for Identifying Citrulline-specific Monoclonal Antibodies from Single Human B Cells from Rheumatoid Arthritis Patient Material

Authors: Khaled Amara, Lena Israelsson, Ragnhild Stålesen, Peter Sahlström, Johanna Steen, Vivianne Malmström and Caroline Grönwall, date: 09/05/2019, view: 1302, Q&A: 0

[Abstract] We describe here a detailed, refined protocol for the generation of citrulline-specific monoclonal antibodies from single human B cells from rheumatoid arthritis (RA) patients. This protocol provides a detailed guide of the procedure starting from single B cells of your choice and followed by amplification of the variable region of immunoglobulin ...

Production and Purification of Dengue Virus-like Particles from COS-1 Cells

Authors: Jedhan Ucat Galula, Gwong-Jen J. Chang and Day-Yu Chao, date: 06/20/2019, view: 2533, Q&A: 0

[Abstract] Non-infectious virus-like particles (VLPs) containing dengue virus (DENV) pre-membrane (prM) and envelope (E) proteins have been demonstrated to be highly immunogenic and can be used as a potential vaccine candidate as well as a tool for serodiagnostic assays. Successful application of VLPs requires abundant, and high-purity production methods. ...

Quantification of Serum Ovalbumin-specific Immunoglobulin E Titre via in vivo Passive Cutaneous Anaphylaxis Assay

Authors: Kyle T Mincham, Jonatan Leffler, Naomi M Scott, Jean-Francois Lauzon-Joset, Philip A Stumbles, Patrick G Holt and Deborah Strickland, date: 03/05/2019, view: 2053, Q&A: 0

[Abstract] Murine models of allergic airway disease are frequently used as a tool to elucidate the cellular and molecular mechanisms of tissue-specific asthmatic disease pathogenesis. Paramount to the success of these models is the induction of experimental antigen sensitization, as indicated by the presence of antigen-specific serum immunoglobulin E. The ...

Characterization of Mouse Adult Testicular Macrophage Populations by Immunofluorescence Imaging and Flow Cytometry

Authors: Noushin Mossadegh-Keller and Michael H. Sieweke, date: 03/05/2019, view: 1988, Q&A: 0

[Abstract] Testicular macrophages (tMΦ) are the most abundant immune cells residing in the testis, an immune-privileged organ. TMΦ are known to exhibit different functions, such as protecting spermatozoa from auto-immune attack by producing immunosuppressive cytokines and trophic roles in supporting spermatogenesis and male sex hormone production. They also ...

DNA Immunization Using in vivo Electroporation for Generating Monoclonal Antibodies Against Mouse IL-9R

Authors: Shogo Takatsuka, Hiroyuki Yamada, Kei Haniuda, Marina Ichihashi, Joe Chiba and Daisuke Kitamura, date: 02/20/2019, view: 2169, Q&A: 0

[Abstract] Membrane proteins such as cytokine receptors and G protein-coupled receptors can be drug targets. Recently, we have generated specific monoclonal antibodies (mAbs) against the mouse IL-9 receptor (IL-9R) and found that IL-9R on memory B cells have critical roles in T-dependent immune response. So far, most antibodies against cell surface proteins ...

Electroporation of Labeled Antibodies to Visualize Endogenous Proteins and Posttranslational Modifications in Living Metazoan Cell Types

Authors: Sascha Conic, Dominique Desplancq, László Tora and Etienne Weiss, date: 11/05/2018, view: 2419, Q&A: 0

[Abstract] The spatiotemporal localization of different intracellular factors in real-time and their detection in live cells are important parameters to understand dynamic protein-based processes. Therefore, there is a demand to perform live-cell imaging and to measure endogenous protein dynamics in single cells. However, fluorescent labeling of endogenous ...

Nab Escaping AAV Mutants Isolated from Mouse Muscles

Authors: Zheng Chai, R. Jude Samulski and Chengwen Li, date: 05/05/2018, view: 3476, Q&A: 0

[Abstract] Neutralizing antibodies (Nabs) are a major challenge in clinical trials of adeno-associated virus (AAV) vector gene therapy, because Nabs are able to inhibit AAV transduction in patients. We have successfully isolated several novel Nab-escaped AAV chimeric capsids in mice by administrating a mixture of AAV shuffled library and patient serum. These ...

Qualitative and Quantitative Assay for Detection of Circulating Autoantibodies against Human Aortic Antigen

Authors: Brent Veerman and Ritu Chakravarti, date: 07/05/2017, view: 5955, Q&A: 0

[Abstract] Increased amount of autoantibodies in human sera are the hallmark of autoimmune diseases (Wang et al., 2015). In case of known antigen, detection of autoantibodies is done using laboratory based methods. However, in most autoimmune diseases, knowledge of self-antigen is still vague. We have developed an ELISA-based quantitative assay to ...

Generation of IgG-Fc Glycovariants Using Recombinant Glycosidases and Glycosyltransferases

Authors: Isaak Quast, Michael A. Maurer and Jan D. Lünemann, date: 08/05/2016, view: 8253, Q&A: 0

[Abstract] The immunoglobulin G (IgG) fragment crystallizable (Fc) domain contains a single, highly conserved asparagine 297 (N297) glycosylation site in the CH2 domain, which is buried within the hydrophobic core of each of the two heavy chains. The biantennary core glycan structure, composed of 2 N-acetylglucosamine (GlcNAc) and 3 mannose residues, can be ...

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