Flashcards in Immunoassays Deck (16):
many B cells respond to antigen, antibodies to many different antigen in serum, multiple specificities and affinities
purify antibody you want
single clone of one B cell is produced by fusing that cell to a tumor cell and isolating the clone with the antibody specificity of interest. (drugs can now target specific antigen)
produces population of antibody molecules that are absolutely identical with same specificity. there is no heterogeneity.
immortal and monoclonal, result of a fusion between B cells and myeloma (tumor cells that are immortal)
hypersensitivity reaction, immune system reaction to antiserum, can result in fever, rash, swollen lymph nodes
ELISA enzyme linked immunosorbant assay
most common assay (ex. home pregnancy test), secondary antibody linked with enzyme is used that changes color, the more significant the color change, the more antibody/enzyme is bound to it (increase concentration)
coat plastic wells with antibody to IL-2. add solution of IL-2. wash away unbound IL-2. monitor conc of IL-2 by adding secondary antibody linked with enzyme that binds to first antibody.
used to assay for antibody or antigen
mixture of cells labeled with fluorescent antibodies is sent through a stream and is counted by detector
determines apparent molecular weight and concentration of antigen, separates protein by size, uses antibody to see if there is a specific protein
electric current, separates proteins by size, similar to ELISA, chemical agent glows in dark if enzyme is present.
will kill you before your body develops antibodies to it
neutralized toxin that is used in vaccines
affinity purify antibody
makes the antibody more specific by purifying the antibody you want
absorb the cross reacting antibody
makes the antibody more specific by removing the contaminating antibody
part of the variable region, do not vary much from one protein to another, forms the bulk of the variable region
provide sequence of the very specific antigen binding site, part of the variable region, forms the very tip of the variable region that binds to antigen
preparation of monoclonal antibodies
immunize animal, isolate spleen cells, fuse spleen cells with tumor cells, select hybrids of tumor and b cells, clone hybridomas, select individual clone with specificity you are interested in