Hybridoma & Vaccines CS Flashcards
what is hybridoma technology
one of the technique used to produce monoclonal antibodies
what is the antibodies that are generated by immunization
are polyclonal antibodies.
with different specificities, affinities and properties
how to solve above problem
create ab with homogeneous structure, same specificity and affinity. that will be monoclonal ab
what is the difference btn monoclonal and polyclonal antibody
monoclonal antibody are antibody produced by one B cells that recognize only one epitope of antigen
polyclonal antibody present different antibodies that are produced by different B cells and can recognize multiple epitope of same antigen
what are strategies of producing monoclonal antibody
- preparation of immunogen
- immunization
- early testing
- cloning and antibody production
- purification
- quality control
how is immunization done
you first inject immunogen / target antigen in mouse and isolate antibody producing cells (B cells) from spleen of mouse
how is immortalization done
is fusion of immortalized myeloma cell line and B cells via PEG fusion. cell will produce ab and divide indefinity
how is HAT selection work
for cell to grow in HAT they have to be positive FOR HGPRT enzyme. B cells can grow in HAT but myeloma cell can’t.
normal cells tolerate HAT but can’t survive in culture
what is the ability of fused hybridoma cells
hybridoma cells are HGPRT+, they are able to survive in culture and even in the presence of HAT media
how cloning base on detected ab is done
you first select hybridoma that produce ab of desired specificity to antigen. then production of monoclonal ab
answer this questions
- What is the difference between an antigen and an immunogen?
- What is the difference between antisera technology and hybridoma technology?
- What are the six main strategy points to be followed for the production of a monoclonal
antibody (mAb)? Mention and elaborate on them. - Why can’t we use B cells alone to create a mAb?
* B cells can’t survive in the culture
* B cells will produce polyclonal antibodies - What is the importance of using myeloma cells that lack the enzyme HGPRT?
* to help hybridoma cells to survive in culture
Vaccines & vaccination
what is vaccine
is biological product that induce immune response for protection
what does vaccine have to contain
it have to contain antigen either from pathogen or produced one to represent component of antigen
why do we need vaccine
to protect us against diseases
