Immunology Lecture 3. Flashcards
What is the equation for measuring antibody affinity?
K = (Ab Ag) / (Ab) (Ag) —> (antibody bound to antigen / non bound antibody x non bound antigen)
What is an epitope?
the part of protein that is actually bound, for example, the hypervariable region is the epitope of an antibody
What is avidity?
the strength of binding of multivalent antiserum to multivalent antigen
What are the steps in producing a monoclonal antibody?
- immunize an animal 2. isolate spleen cells 3. fuse spleen cells to melanoma cells 4. select for hybrids 5. clone the hybridomas so that each single cell grows up independently 6. select the individual clone with the specificity you are interested in
Why are monoclonal antibodies so important?
we can make huge amounts of a specific antibody - there will be NO heterogeneity
What is serum sickness?
immune response to injected proteins
How do you know if a drug is a monoclonal antibody?
ends with -mab
What is enzyme-linked immunosorbent assay
ELISA - antigen stuck to bottom of tube - antibody added, second antibody binds to first, amount of second antibody is detected by chemical reagent that turns color - more enzyme = more color change - a way of screening blood from an antigen (ex. HIV)
What is immunofluorescence?
tissue/cells reacted with antisera specific for a cell maker or pathogen, second antibody added which has an attached fluorescent
What is flow cytometry?
a machine that scans large numbers of cells for immunofluorescence as well as cell size (graphed on a two dimensional dot plot)
What is western immunoblot?
separate PROTEIN by weight and concentration - quantitative and qualitative
What can you learn about an antigen using a western immunoblot but not an ELISA?
molecular weight
