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Protocol for Recombinant RBD-based SARS Vaccines: Protein Preparation, Animal Vaccination and Neutralization Detection
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Protocol for Recombinant RBD-based SARS Vaccines: Protein Preparation, Animal Vaccination and Neutralization Detection

Protocol for Recombinant RBD-based SARS Vaccines: Protein Preparation, Animal Vaccination and Neutralization Detection

DOI:
10.3791/2444-v

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12:09 min

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May 02, 2011

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, , ,
1Lindsley F. Kimball Research Institute,New York Blood Center

Chapters

  • 00:05Title
  • 01:01Introduction
  • 01:52Recombinant SARS-CoV RBD Protein Preparation
  • 04:40Mouse Immunization and Sample Collection
  • 05:56Preparation of SARS Pseudovirus in 293T Cells
  • 06:53SARS Pseudovirus Titration and Neutralization Assay
  • 11:10Evaluation of Recombinant RBD-induced Neutralizing Activity
  • 11:36Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol describes a general procedure for studying recombinant receptor-binding domain (RBD)-based subunit vaccines against SARS. It includes methods for transfection and expression of RBD protein in 293T cells, immunization of mice with RBD and detection of neutralization activity of mouse sera using an established SARS pseudovirus neutralization assay.

Tags

Recombinant RBD-based SARS VaccinesProtein PreparationAnimal VaccinationNeutralization DetectionSubunit VaccinesMammalian Cell SystemAntigenicityImmunogenicitySARS-CoVRBD ProteinReceptor BindingNeutralizing AntibodiesCulture SupernatantMammalian 293T CellsTransfection MethodCalcium Phosphate Transfection

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