Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Antigen Processing Pathways01:31

Antigen Processing Pathways

2.0K
MHC molecules are key players in the immune response, enabling T cells to recognize and respond to specific antigens. They are present on the surface of all nucleated cells in the body and are instrumental in presenting antigens to T cells and activating them. T cells recognize the MHC-antigen complex and initiate an immune response. MHC class I and MHC class II are two main types of MHC molecules, each associated with a distinct antigen processing pathway.
MHC Class I: Presenting Endogenous...
2.0K
Antigens Involved in Adaptive Immunity01:26

Antigens Involved in Adaptive Immunity

1.2K
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...
1.2K
T Cell Activation and Clonal Selection01:22

T Cell Activation and Clonal Selection

14.5K
T cells are integral to our adaptive immune system, recognizing and effectively responding to foreign antigens. T cell activation and clonal selection are pivotal in orchestrating this immune response. This article elucidates these mechanisms, detailing the roles of cluster of differentiation (CD) markers, major histocompatibility complex (MHC) molecules, costimulatory signals, and the process of clonal selection.
Naive T cells that have not yet encountered an antigen express two primary CD...
14.5K
Hybridoma Technology01:31

Hybridoma Technology

17.1K
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,...
17.1K
Cell-mediated Immune Responses01:40

Cell-mediated Immune Responses

83.3K
Overview
83.3K
B Cell Activation and Differentiation01:24

B Cell Activation and Differentiation

15.8K
The adaptive immune response, a sophisticated defense mechanism, relies on the activation and differentiation of B lymphocytes, or B cells. These processes enable our bodies to mount a tailored response against specific pathogens such as bacteria, free virus particles, toxins, and parasites.
When naive B cells encounter a specific antigen that can bind to the B cell receptor (BCR) on their surface, they undergo sensitization to respond to the antigen's presence. Sensitization begins with...
15.8K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

How conformational changes near the F pocket of MHC class I proteins mediate chaperone assisted peptide loading.

Frontiers in immunology·2025
Same author

Surface emergence and persistence of MHC class I free heavy chains.

The Journal of biological chemistry·2025
Same author

X-ray spectroscopy meets native mass spectrometry: probing gas-phase protein complexes.

Physical chemistry chemical physics : PCCP·2025
Same author

New insights into the immunomodulatory potential of sialic acid on monocyte-derived dendritic cells.

Cancer immunology, immunotherapy : CII·2024
Same author

Peptide vaccines get an OS update.

Nature chemical biology·2024
Same author

Expanding the MAPPs Assay to Accommodate MHC-II Pan Receptors for Improved Predictability of Potential T Cell Epitopes.

Biology·2023

Related Experiment Video

Updated: Jan 7, 2026

Stability and Structure of Bat Major Histocompatibility Complex Class I with Heterologous β2-Microglobulin
11:17

Stability and Structure of Bat Major Histocompatibility Complex Class I with Heterologous β2-Microglobulin

Published on: March 10, 2021

6.7K

Empty MHC Class I Protein Production.

Ankur Saikia1, Yelyzaveta Makedon1, Bianka Nagy1

  • 1Constructor University, Bremen, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|January 1, 2026
PubMed
Summary

This study details a protocol for producing and refolding peptide-empty MHC class I proteins. This method yields functional proteins for biochemical research.

Keywords:
Disulfide-mediated stabilizationE. coli expressionInclusion bodiesMHC Class IProtein refoldingSize exclusion chromatography

More Related Videos

Use of Single Chain MHC Technology to Investigate Co-agonism in Human CD8+ T Cell Activation
12:09

Use of Single Chain MHC Technology to Investigate Co-agonism in Human CD8+ T Cell Activation

Published on: February 28, 2019

10.2K
Assessing the Expression of Major Histocompatibility Complex Class I on Primary Murine Hippocampal Neurons by Flow Cytometry
08:07

Assessing the Expression of Major Histocompatibility Complex Class I on Primary Murine Hippocampal Neurons by Flow Cytometry

Published on: May 19, 2020

3.8K

Related Experiment Videos

Last Updated: Jan 7, 2026

Stability and Structure of Bat Major Histocompatibility Complex Class I with Heterologous β2-Microglobulin
11:17

Stability and Structure of Bat Major Histocompatibility Complex Class I with Heterologous β2-Microglobulin

Published on: March 10, 2021

6.7K
Use of Single Chain MHC Technology to Investigate Co-agonism in Human CD8+ T Cell Activation
12:09

Use of Single Chain MHC Technology to Investigate Co-agonism in Human CD8+ T Cell Activation

Published on: February 28, 2019

10.2K
Assessing the Expression of Major Histocompatibility Complex Class I on Primary Murine Hippocampal Neurons by Flow Cytometry
08:07

Assessing the Expression of Major Histocompatibility Complex Class I on Primary Murine Hippocampal Neurons by Flow Cytometry

Published on: May 19, 2020

3.8K

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Immunology

Background:

  • MHC class I proteins are crucial for immune response.
  • Recombinant production of functional MHC class I proteins is challenging.
  • Disulfide-stabilized, peptide-empty forms are valuable research tools.

Purpose of the Study:

  • To provide a detailed protocol for recombinant production of MHC class I proteins.
  • To describe in vitro refolding of disulfide-stabilized, peptide-empty MHC class I molecules.
  • To enable the generation of functional MHC class I proteins for biochemical and biophysical studies.

Main Methods:

  • Expression of MHC class I heavy and light chains (β2m) in E. coli as inclusion bodies.
  • Purification of expressed proteins and inclusion bodies.
  • In vitro refolding of disulfide-stabilized MHC class I molecules using a dilution method with dipeptides.

Main Results:

  • Successful recombinant production of MHC class I heavy and light chains.
  • Efficient purification of inclusion bodies containing the target proteins.
  • Achieved in vitro refolding of disulfide-stabilized, peptide-empty MHC class I proteins using a dilution method.

Conclusions:

  • The protocol effectively produces correctly folded, functional, peptide-empty MHC class I proteins.
  • This method is suitable for generating proteins required for various biochemical and biophysical assays.
  • The developed protocol facilitates research in immunology and protein structure-function relationships.