Antibody-antigen interaction —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: 4400, 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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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: 3019, 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 ...

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: 6812, 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. ...

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: 4974, 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 ...

Kinetic Analysis of Monoclonal Antibody Binding to HIV-1 gp120-derived Hyperglycosylated Cores

Authors: Jidnyasa Ingale and Richard T Wyatt, date: 10/05/2015, view: 7346, Q&A: 0

[Abstract] Kinetic analysis of antibodies is one of the important studies for characterization of antibodies and screening of ligands. In our recent study (Ingale et al., 2014), we compared the antigenic profiles and binding characteristics of four HIV-1 envelope glycoprotein (Env) core immunogens using multiple monoclonal antibodies by Bio-Layer ...

Fluorescence-linked Antigen Quantification (FLAQ) Assay for Fast Quantification of HIV-1 p24^Gag

Authors: Marianne Gesner, Mekhala Maiti, Robert Grant and Marielle Cavrois, date: 12/20/2014, view: 10102, Q&A: 1

[Abstract] The fluorescence-linked antigen quantification (FLAQ) assay allows a fast quantification of HIV-1 p24^Gag antigen. Viral supernatant are lysed and incubated with polystyrene microspheres coated with polyclonal antibodies against HIV-1 p24^Gag and detector antibodies conjugated to fluorochromes (Figure 1). After washes, the ...

Binding Affinity Measurement of Antibodies from Crude Hybridoma Samples by SPR

Authors: Dorin Mlaki Ndao, David T. Hickman, María Pilar López-Deber, Aurélien Davranche, Andrea Pfeifer and Andreas Muhs, date: 11/05/2014, view: 14505, Q&A: 0

[Abstract] Surface Plasmon Resonance (SPR) is widely used to generate kinetic and affinity information on specific interactions between biomolecules. This technique is label-free and monitors the binding event in real-time. It is generally used for characterization of monoclonal antibody - antigen interactions. This protocol describes specifically the use of ...

α2β1-integrin Clustering and Internalization Protocol

Authors: Varpu Marjomäki, Mikko Karjalainen, Paula Upla, Elina Siljamäki, Nina Rintanen and Paula Turkki, date: 04/05/2014, view: 8442, Q&A: 0

[Abstract] α2β1-integrin clustering experiment can be used to trigger internalization of α2β1-integrin. When clustering is performed with sequential administration of primary and fluorescent secondary antibodies, the entry kinetics of integrin can be followed into the cell. The idea is first to allow binding of primary antibodies (recognizing the ...

Chikungunya ELISA Protocol

Authors: Caitlin Mattos Briggs, Amanda Piper and Raquel Hernandez, date: 02/20/2014, view: 9021, Q&A: 0

[Abstract] This protocol will result in the accurate qualitative measurement of anti-Chikungunya virus antibody (Ab) from infected mouse tissue or serum. This assay was developed by Dr. Caitlin Briggs, Arbovax, Inc. Chikungunya is a BL3 agent and should be handled in a biosafety level 3 laboratory under BL3 conditions. This protocol was used in the ...

Surface Plasmon Resonance Analysis of Antigen-Antibody Interaction

Authors: Mitsuhiro Nishimura and Kazuyoshi Ikuta, date: 12/20/2013, view: 11993, Q&A: 0

[Abstract] Molecular interaction between monoclonal antibodies (MAbs) and their recognized antigen is a fundamental event leading to the neutralization activity. Estimation of their binding affinity gives beneficial information to characterize the MAbs and to develop more effective MAbs. Surface plasmon resonance (SPR) analysis is a powerful tool to analyze ...

Immunology

Protocol for Isolation of Cardiomyocyte from Adult Mouse and Rat

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