Antibody analysis —Immunology

Production of the Receptor-binding Domain of the Viral Spike Proteins from 2003 and 2019 SARS CoVs and the Four Common Human Coronaviruses for Serologic Assays and Inhibitor Screening

Authors: Bruno Segovia-Chumbez, Stephen D. Graham, Ramesh Jadi, Aravinda M. de Silva and Lakshmanane Premkumar, date: 05/20/2021, view: 3193, Q&A: 0

[Abstract]

The recombinant receptor-binding domain (RBD) of the viral spike protein from SARS-CoV-1 and 2 are reliable antigens for detecting viral-specific antibodies in humans. We and others have shown that the levels of RBD-binding antibodies and SARS-CoV-2 neutralizing antibodies in patients are correlated. Here, we report the expression and purification

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An Image-based Dynamic High-throughput Analysis of Adherent Cell Migration

Authors: Meng Sun, Bence Rethi, Akilan Krishnamurthy, Vijay Joshua, Heidi Wähämaa, Sergiu-Bogdan Catrina and Anca Catrina, date: 03/20/2021, view: 2721, Q&A: 0

[Abstract]

In this protocol, we describe a method to monitor cell migration by live-cell imaging of adherent cells. Scratching assay is a common method to investigate cell migration or wound healing capacity. However, achieving homogenous scratching, finding the optimal time window for end-point analysis and performing an objective image analysis imply, even

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Multiplication and Growth Inhibition Activity Assays for the Zoonotic Malaria Parasite, Plasmodium knowlesi

Authors: Franziska Mohring, Thomas A. Rawlinson, Simon J. Draper and Robert W. Moon, date: 09/05/2020, view: 2483, Q&A: 0

[Abstract] Malaria remains a major cause of morbidity and mortality globally. Clinical symptoms of the disease arise from the growth and multiplication of Plasmodium parasites within the blood of the host. Thus in vitro assays to determine how drug, antibody and genetic perturbations affect the growth rate of Plasmodium parasites ...

Assessments of HLA-I Specificities of Anti-HLA-I Monoclonal Antibodies Using Solid Phase Bead Arrays

Authors: Anita J. Zaitouna, Daniel S. Ramon and Malini Raghavan, date: 06/20/2020, view: 1931, Q&A: 0

[Abstract] Human leukocyte antigen class I (HLA-I) molecules are a group of structurally-related cell surface proteins with a high degree of variability within the population. While only up to six variants are expressed in an individual person, the whole population contains thousands of different variants. The ability to distinguish specific variants is ...

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: 3219, 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: 2999, 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: 5568, 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: 3663, 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: 3770, 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: 4160, 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 ...

Immunology

Antisense Oligo Pulldown of Circular RNA for Downstream Analysis

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