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Determination of the Relative Potency of an Anti-TNF Monoclonal Antibody (mAb) by Neutralizing TNF Using an In Vitro Bioanalytical Method
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JoVE Journal Medicina
Determination of the Relative Potency of an Anti-TNF Monoclonal Antibody (mAb) by Neutralizing TNF Using an In Vitro Bioanalytical Method

Determination of the Relative Potency of an Anti-TNF Monoclonal Antibody (mAb) by Neutralizing TNF Using an In Vitro Bioanalytical Method

DOI:
10.3791/55376-v

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16:07 min

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September 16, 2017

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

Capítulos

  • 00:00Título
  • 00:31Introduction
  • 02:32Methodology: Solutions
  • 02:51Methodology: Cells Thawing and First Subculturing
  • 04:33Methodology: Verification of Confluency
  • 05:21Methodology: Second Subculturing, Cells Detaching and Counting
  • 07:45Methodology: Antibody Quantitation and Dilutions
  • 09:37Methodology: Neutralization Assay
  • 13:00Methodology: Spectrophotometric Analysis
  • 13:54Results: Dose Response Curve
  • 15:23Conclusion

Summary

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Tadução automática

A protocol for the determination of the relative anti-apoptotic activity of an anti-TNFα mAb using a neutralization mechanism with WEHI 164 cells is presented here. This protocol is useful for comparing the neutralization strength of different molecules with the same biological functionality.

Tags

Keywords: Anti-TNF Monoclonal AntibodyTNF-alpha NeutralizationIn Vitro Bioanalytical MethodRelative PotencyWEHI 164 CellsApoptosisCell CultureCell DensityAntibody DilutionSpectrophotometric Analysis

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