Immunoassays

Question 1

polyclonal

Answer 1

many B cells respond to antigen, antibodies to many different antigen in serum, multiple specificities and affinities

Question 2

affinity purify

Answer 2

purify antibody you want

Question 3

monoclonal antibodies

Answer 3

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.

Question 4

hybridoma

Answer 4

immortal and monoclonal, result of a fusion between B cells and myeloma (tumor cells that are immortal)

Question 5

serum sickness

Answer 5

hypersensitivity reaction, immune system reaction to antiserum, can result in fever, rash, swollen lymph nodes

Question 6

ELISA enzyme linked immunosorbant assay

Answer 6

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

Question 7

flow cytometry

Answer 7

mixture of cells labeled with fluorescent antibodies is sent through a stream and is counted by detector

Question 8

western blot

Answer 8

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.

Question 9

tetanus toxins

Answer 9

will kill you before your body develops antibodies to it

Question 10

tetanus toxoid

Answer 10

neutralized toxin that is used in vaccines

Question 11

affinity purify antibody

Answer 11

makes the antibody more specific by purifying the antibody you want

Question 12

absorb the cross reacting antibody

Answer 12

makes the antibody more specific by removing the contaminating antibody

Question 13

framework residues

Answer 13

part of the variable region, do not vary much from one protein to another, forms the bulk of the variable region

Question 14

hypervariable region

Answer 14

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

Question 15

preparation of monoclonal antibodies

Answer 15

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

Question 16

immunofluorescence

Answer 16

can be used to identify a specific cell type or find a specific cellular structure. It may also be used to identify a pathogen within cells or tissue