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Related Concept Videos

Antibody Actions01:26

Antibody Actions

Antibodies, or immunoglobulins, are critical players in the immune system's arsenal against invading pathogens. Produced by B cells and plasma cells, their primary role is to detect and bind to specific antigens, molecules found on the surface of pathogens like bacteria or viruses. Beyond antigen recognition, antibodies perform several vital functions that contribute to immune defense.
Neutralization
Antibodies can bind to pathogens, preventing them from infecting host cells. This process...
Immunoprecipitation01:20

Immunoprecipitation

Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
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...
Affinity and Avidity01:41

Affinity and Avidity

Overview
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...
Antigens Involved in Adaptive Immunity01:26

Antigens Involved in Adaptive Immunity

An antigen is any substance the immune system identifies as foreign and potentially harmful to the body, prompting an immune response. Antigens have two functional properties: immunogenicity and reactivity. Immunogenicity is the ability of an antigen to stimulate a specific immune response. At the same time, reactivity describes the antigen's ability to react with the cells and antibodies produced in response to it.
Complete Antigens
Complete antigens possess both immunogenicity and reactivity.

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Updated: Jun 20, 2026

Initial Evaluation of Antibody-conjugates Modified with Viral-derived Peptides for Increasing Cellular Accumulation and Improving Tumor Targeting
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Initial Evaluation of Antibody-conjugates Modified with Viral-derived Peptides for Increasing Cellular Accumulation and Improving Tumor Targeting

Published on: March 8, 2018

Antibody internalization after cell surface antigen binding is critical for immunotoxin development.

Shu-Ru Kuo1, Randall W Alfano, Arthur E Frankel

  • 1Cancer Research Institute of Scott & White Hospital and Department of Internal Medicine, Health Science Center, College of Medicine, Texas A&M University, 5701 South Airport Road, Temple, TX 76502, USA.

Bioconjugate Chemistry
|September 30, 2009
PubMed
Summary

This study introduces a new assay using DTG3 to evaluate antibody internalization efficiency for developing effective immunotoxins. This method helps select antibodies that efficiently target cancer cells for improved therapeutic outcomes.

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Detection of Antibodies That Neutralize the Cellular Uptake of Enzyme Replacement Therapies with a Cell-based Assay
07:52

Detection of Antibodies That Neutralize the Cellular Uptake of Enzyme Replacement Therapies with a Cell-based Assay

Published on: September 10, 2018

Area of Science:

  • Immunology
  • Molecular Biology
  • Oncology

Background:

  • Immunotoxin efficacy relies on antibody binding specificity and cellular internalization.
  • Existing antibody selection methods prioritize affinity and yield, potentially overlooking internalization efficiency crucial for immunotoxin design.

Purpose of the Study:

  • To develop and validate a novel assay for comparing monoclonal antibody internalization efficiency.
  • To identify optimal antibodies for immunotoxin development, overcoming limitations of current selection strategies.

Main Methods:

  • A novel fusion protein, DTG3 (diphtheria toxin N-terminus and Protein G domains), was engineered.
  • Antibody-DTG3 complexes were formed, tested for cell surface binding, internalization, and diphtheria toxin-mediated cytotoxicity.
  • The assay was validated using anti-CD3ε and anti-PSMA antibodies and applied to screen anti-CD123 antibodies for leukemia-targeting immunotoxins.

Main Results:

  • The DTG3 assay successfully demonstrated antibody-mediated cell internalization and subsequent apoptosis.
  • The system proved effective in evaluating known antibodies and screening novel hybridomas for immunotoxin development.
  • Commercial anti-CD123 antibodies were assessed for potential in leukemia-targeting immunotoxin therapies.

Conclusions:

  • The developed DTG3-based assay is sensitive, efficient, and reproducible for comparing antibody internalization.
  • This assay provides a valuable tool for selecting optimal antibodies in the early stages of immunotoxin development.
  • The findings facilitate the clinical development of more effective immunotoxins for treating various neoplasms.