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Related Concept Videos

Vaccine Production01:23

Vaccine Production

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Vaccine production involves a sequence of upstream and downstream processes to generate a safe and effective immunological product. It begins with cultivating microorganisms, such as viruses or bacteria, to obtain antigenic material. For viral vaccines, mammalian host cells are grown in bioreactors and subsequently infected with the target virus. The virus replicates within the host cells, which are lysed to release viral particles. This lysate is then clarified through filtration or...
136

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Related Experiment Video

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Protocol for Recombinant RBD-based SARS Vaccines: Protein Preparation, Animal Vaccination and Neutralization Detection
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Engineering a SARS-CoV-2 vaccine targeting the RBD cryptic-face via immunofocusing.

Theodora U J Bruun1,2, Jonathan Do1,2, Payton A-B Weidenbacher1,3

  • 1Sarafan ChEM-H, Stanford University, Stanford, CA 94305.

Biorxiv : the Preprint Server for Biology
|June 19, 2024
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Summary
This summary is machine-generated.

New vaccine strategies using PMD-RBD immunogens successfully focused antibody responses to target the SARS-CoV-2 spike protein's cryptic face. This approach enhances broad neutralization against Sarbecoviruses, crucial for future vaccine development.

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Area of Science:

  • Virology
  • Immunology
  • Vaccine Development

Background:

  • The SARS-CoV-2 spike protein's receptor-binding domain (RBD) is a key target for neutralizing antibodies.
  • Antibodies targeting the RBD's cryptic face show broad neutralization across Sarbecoviruses but are rarely elicited naturally.

Purpose of the Study:

  • To develop novel receptor-binding domain (RBD) immunogens using the PMD (protect, modify, deprotect) technique.
  • To focus the antibody response towards the cryptic face epitope of the RBD, recognized by broadly neutralizing antibodies like S2X259.

Main Methods:

  • Employed the immunofocusing technique PMD to create specifically designed PMD-RBD immunogens.
  • Administered PMD-RBD antigens to induce immune responses.
  • Utilized serum-depletion assays to analyze the polyclonal antibody response.

Main Results:

  • PMD-RBD immunization induced high binding titers and broad neutralizing activity against various Sarbecovirus strains.
  • Serum-depletion assays confirmed that PMD successfully directed the antibody response towards the RBD's cryptic face.
  • Demonstrated the ability of PMD to overcome immunodominance and refocus humoral immunity.

Conclusions:

  • The PMD technique effectively focuses antibody responses to cryptic epitopes on the SARS-CoV-2 RBD.
  • PMD-RBD immunogens offer a promising strategy for developing broader and more resilient vaccines against Sarbecoviruses and emerging viral threats.