In my research I would like to find effective antigens that will be able to be included in vaccines. My means of research are human PBMCs from healthy donors and a series of in vitro assays.
After a few literature reading it is not clear to me which can be the key experiments/assays that I can perform to demonstrate the immunogenicity of these antigens and that they can be good candidates to be included in future vaccines.
Do you have any experience in this or can suggest me some interesting reading?
In the same way the APCs from the antigens, you could this in vitro. Maybe use this as a technique. Sorry I can't provide any literature for this.
It doesn't seem right that you have an idea of specimen/materials to use for your study before having an adequate phrasing of your objectives, hypotheses and strategy for interrogating them. If you are planning an exploratory work for a topic or vaccine candidate lead that has not been pursued yet, consider doing an in silico approach first:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7120937/
https://www.nature.com/articles/s41598-020-79645-9
https://www.nature.com/articles/s41598-020-63640-1
A straightforward, but definitely crude, way of preliminary assessment of immunogenicity of a certain candidate is to introduce them (and their possible chemical modifications or additions) to animals such as mice and rabbits and ascertain later on if they develop binding antibodies against them (remember, though, the immunity is not limited to antibody-based responses).
Normally for Showing immunogenicity cell culture based systems is ideal.
TCID-50, tissue culture infectivity dose 50. we culture cells and plate them in a 96 well plate and then add virus and observe the CPE cytopathic effect. if some component is protecting this can be added and see how much this reduces the TCID-50 values.
Serum neutralization assays are perfomed to check the protection from the infecting viurs. SNT assay. when monoclonal antibodies are used for testing protection instead of serum monoclonal antibodies can be used.
OIE is for animal health we can search vaccines. they give all standard protocols.
if it shows good cytopathic effect then same can be injected into animals for invo testing. the protection shown in cell culture system can be replicated in animals also.
the viruses infect prefentially so we need to choose the cell lines accordingly. eg for SARS- CoV2 they use Vero cell line, for FMDV they used BHK21Cl13 cell line including for vaccine BHK Raji cell line is used. for Infecious Bovine Rhinotracheitis.
learning cell culture and having a set up with laminar flow with sterile facility is needed, little bit can be done in laminar flow.
Mª Angeles Zorrilla Lopez-Perea Aedrian Abrilla Michael Durairaj S
Thank you all for your replies!
https://www.oie.int/doc/ged/D7722.PDF
This has for animal viruses, similarly WHO should have one for human viruses but method of checking infectivity and protection is the same.
Immunogens viral,bactrial or parasitc and toxoid.
Viral vaccine development in vitro tissue vaccine,emryonated egg vaccine,and these divided into :A Inactivated vaccine(killed) by different inactivating agents like
formalin,bet_propiolactone,E imen,Methyl ethyel emin and formalin.
B: Attenuated vaccine using different passages on tissue culture,Embryonated Eggs and lab animals which leads lost it,s virulance not immunogenicity.
About bacterial vaccine the same also killed or avirulant vaccine or their product toxoid.My suggestion for science of vaccinology you must focus on your aim:
Immunogen,adjuvant and inactivating agent.
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