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Détermination de la puissance Relative d’un anticorps Monoclonal Anti-TNF (mAb) de neutralisants ...
Détermination de la puissance Relative d’un anticorps Monoclonal Anti-TNF (mAb) de neutralisants ...
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JoVE Journal Medicine
Determination of the Relative Potency of an Anti-TNF Monoclonal Antibody (mAb) by Neutralizing TNF Using an In Vitro Bioanalytical Method

Détermination de la puissance Relative d’un anticorps Monoclonal Anti-TNF (mAb) de neutralisants TNF en utilisant une méthode de Bioanalytical In Vitro

Full Text
9,701 Views
16:07 min
September 16, 2017

DOI: 10.3791/55376-v

Lilia Tierrablanca-Sánchez1, Víctor Pérez Medina Martínez1, Nancy D. Ramírez Ibañez1, Néstor O. Pérez Ramírez1, Francisco Luis Flores Ortiz1, Emilio Medina-Rivero2

1Research and Development,Probiomed, 2Bioprocess Research and Development Unit,Instituto Politecnico Nacional

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This protocol outlines a method for assessing the relative potency of anti-TNF-alpha monoclonal antibodies using WEHI 164 cells. It provides a comparative analysis of the neutralization efficiency of a reference mAb against a sample mAb.

Key Study Components

Area of Science

  • Immunology
  • Cell Biology
  • Biotechnology

Background

  • Anti-TNF-alpha monoclonal antibodies are used to neutralize the effects of TNF-alpha.
  • WEHI 164 cells serve as a model for studying cytokine interactions.
  • Understanding neutralization efficiency is crucial for therapeutic applications.
  • Cells can be subjected to stress to evaluate their response to TNF-alpha.

Purpose of Study

  • To measure the relative potency of different anti-TNF-alpha mAbs.
  • To compare the neutralization efficiency of a reference mAb with a sample mAb.
  • To provide a detailed protocol for researchers in the field.

Methods Used

  • Cell culture of WEHI 164 cells.
  • Application of anti-TNF-alpha mAbs to the cultured cells.
  • Assessment of cell viability and apoptosis.
  • Comparison of neutralization potency between mAbs.

Main Results

  • Demonstrated differences in neutralization efficiency between mAbs.
  • Provided quantitative data on the anti-apoptotic activity of tested mAbs.
  • Established a reliable protocol for future studies.
  • Highlighted the importance of receptor interactions in TNF-alpha signaling.

Conclusions

  • The protocol is effective for comparing anti-TNF-alpha mAbs.
  • Understanding neutralization mechanisms can inform therapeutic strategies.
  • Further research is needed to explore additional mAb candidates.

Frequently Asked Questions

What is the significance of TNF-alpha in cell biology?
TNF-alpha is a cytokine involved in systemic inflammation and is a key player in immune responses.
How do anti-TNF-alpha mAbs work?
They bind to TNF-alpha, preventing it from interacting with its receptors, thereby inhibiting inflammatory responses.
What are WEHI 164 cells?
WEHI 164 cells are a murine cell line used for studying immune responses and cytokine interactions.
What factors can stress cells in this protocol?
Environmental factors such as starvation can be used to stress cells and evaluate their response to TNF-alpha.
Why is it important to compare mAbs?
Comparing mAbs helps identify the most effective therapeutic candidates for treating diseases associated with TNF-alpha.

Un protocole pour la détermination de l’activité anti-apoptotique relative d’un anti-TNFα mAb en utilisant un mécanisme de neutralisation avec cellules WEHI 164 est présenté ici. Ce protocole est utile pour comparer la force de la neutralisation des molécules différentes avec les mêmes fonctionnalités biologiques.

L’objectif global de cette procédure est de mesurer la puissance relative des molécules anti-TNF-alpha. Dans cet exemple, nous comparons deux anticorps monoclonaux anti-TNF-alpha afin de déterminer l’efficacité de neutralisation d’un anticorps monoclonal de référence par rapport à un échantillon d’anticorps monoclonal à l’étude. Ce tutoriel montre en détail les étapes de détermination du pouvoir de neutralisation du TNF-alpha, qui ne sont pas incluses dans la littérature.

Les cellules peuvent être stressées par différents facteurs environnementaux, par exemple, la famine. Chaque cellule a différentes protéines à sa surface appelées récepteurs. Certains d’entre eux sont spécifiques de la cytokine TNF-alpha.

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