Related Experiment Video

Updated: May 10, 2026

Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope
08:09

Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope

Published on: March 24, 2017

Polyclonal antibody against conserved peptide in transmembrane protein of avian leukosis virus subgroup J

Guihua Wang1, Xiaowei Wang, Qingqing Xu

  • 1Molecular Pathology Laboratory, Department of Fundamental Veterinary, College of Veterinary Medicine, Shandong Agricultural University, Tai'an, China.

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
|June 12, 2013
PubMed

The ALV-J gp37 pocket region located in transmembrane (TM) protein is an ideal viral target because it is extracellular, highly conserved, and essential for viral entry. Although it has been a target of drug and vaccine design, there are not any polyclonal antibody tools to specifically probe the antigenicity and immunogenicity of the pocket region. Our goal was to elicit a neutralizing polyclonal antibody that targets this pocket region. A conserved peptide designated R73 that is derived from the pocket region of TM of ALV-J was synthesized. New Zealand rabbits were immunized with R73 conjugated with keyhole limpet hemocyanin (KLH), and the antiserum was gained 10 days after the third immunity boost. The antibody titers were tested by endpoint ELISA, and the specificity and affinity were tested by Western blot analysis. The results showed that R73 has good antigenicity (antibody titers≈1:64,000∼1:128,000), immunogenicity, and affinity. The results of the indirect fluorescent assay (IFA) and real-time RT-PCR test showed that R73 polyclonal antibody can recognize ALV-J antigen and completely inhibit viral replication in DF-1 cells. The results of the neutralization assay showed that neutralization titer of the antiserum was 1:16. Therefore, a polyclonal antibody against the neutralizing epitope on an existing pocket region may be a useful tool in ALV-J diagnosis or for preventing infection.

More Related Videos

Generation of Recombinant Arenavirus for Vaccine Development in FDA-Approved Vero Cells10:03

Generation of Recombinant Arenavirus for Vaccine Development in FDA-Approved Vero Cells

Published on: August 1, 2013

Bacterial Artificial Chromosomes: A Functional Genomics Tool for the Study of Positive-strand RNA Viruses12:20

Bacterial Artificial Chromosomes: A Functional Genomics Tool for the Study of Positive-strand RNA Viruses

Published on: December 29, 2015

Related Concept Videos

Hybridoma Technology01:31

Hybridoma Technology

Hybridoma technology is used for the large-scale production of monoclonal antibodies. Monoclonal antibodies bind to only a single antigenic determinant or epitope. Such antibodies are used in research, diagnostics, and disease therapy. The hybridoma technology established in 1975 by Georges Köhler and Cesar Milstein was awarded the Nobel Prize in Medicine in 1984 for revolutionizing research and therapy.
Hybridoma Selection
Commonly used fusion techniques — electroporation, polyethylene glycol...
Antibody Structure01:10

Antibody Structure

Overview
Antibodies, also known as immunoglobulins (Ig), are essential players of the adaptive immune system. These antigen-binding proteins are produced by B cells and make up 20 percent of the total blood plasma by weight. In mammals, antibodies fall into five different classes, which each elicits a different biological response upon antigen binding.
The Y-Shaped Structure of Antibodies Consists of Four Polypeptide Chains
Antibodies consist of four polypeptide chains: two identical heavy...
Antibody Structure01:10

Antibody Structure

Overview
Antibodies, also known as immunoglobulins (Ig), are essential players of the adaptive immune system. These antigen-binding proteins are produced by B cells and make up 20 percent of the total blood plasma by weight. In mammals, antibodies fall into five different classes, which each elicits a different biological response upon antigen binding.
The Y-Shaped Structure of Antibodies Consists of Four Polypeptide Chains
Antibodies consist of four polypeptide chains: two identical heavy...
Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen.  Enzyme-substrate reaction allows the antigen to be visualized or quantified.
Leaky Scanning02:28

Leaky Scanning

During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R stands for...
Immunoglobulin-like Cell Adhesion Molecules01:31

Immunoglobulin-like Cell Adhesion Molecules

Immunoglobulin-like cell adhesion molecules or Ig-CAMs are a versatile group of cell surface glycoproteins belonging to the immunoglobulin protein superfamily. Ig-CAMs possess the characteristic immunoglobulin protein domains and other domains such as the fibronectin type III domain. The Ig domains are glycosylated to varying degrees in different Ig-CAMs.
Ig-CAMs exhibit either homophilic binding (to other Ig-CAMs) or heterophilic binding (to other ligands such as integrins). While most Ig-CAMs...