The molecular biology platform performs PCR-based assays to support vaccine studies. Our qPCR and RT-qPCR assays are used for quantification of the vaccine candidate particles (genome copies) in the samples to support viral shedding studies.
Cell-Mediated Immunity Assays
Vaccine development and evaluations require highly sensitive and quantitative methods that can measure a broad range of molecules such as cytokines, antigenic and cellular markers. Our cellular assays include the following:
Enzyme-linked immunosorbent spot (ELISpot) is a highly flexible and versatile assay that can be adapted to multiple readout formats. ELISpot assays are quantitative and measure key cellular functions of immune system cells. They are useful for assessing both adaptive (humoral) and innate (cell-mediated) immune responses to a vaccine.
Humoral Responses (B-cell)
- Detection of Antibody Secreting Cells (ASC) — IgG/IgA
- Detection of memory B-cells
Cell-Mediated Immune Responses (T-cell)
- Cytokine quantification
- Vaccine efficacy (IFN-γ)
- Quantify immune responses (Th1/Th2)
- Perforin & Granzyme B
- Cytotoxic T-cell responses
In order to identify mediators of short- and/or long-term immunity, a robust method of identification, sorting, and characterization is needed.
Examples of cell types and markers detected using flow cytometry:
- Isolation Peripheral Blood Mononuclear Cells (PBMC) or T-cells (CD4/CD8)
- Intracellular Cytokine Staining (ICS)
- Cytokines (IFN-γ, IL-2, IL-4, IL-5, IL-10, IL-13, IL-17, TNF-α)
- Cytotoxic potential (CD107a, Granzymes, Perforin)
- B-cell help (CD40L)
- Memory profile (CD45RA, CCR7, CD27, CD28)
- Phenotyping (antigen-specific T-cell): ASC (CD3)
- Antibody-Dependent Cellular Cytotoxicity (ADCC) Assays