immuno testing

Question 1

difference between polyclonal and monoclonal antibodies

Answer 1

polyclonal- recognize multiple epitopes, contains various antibodies of different affinities
monoclonal- detect only 1 epitope

Question 2

steps in the production of monoclonal antibody (5)

Answer 2

• isolate spleen cells from mouse that was previously immunized against antigen
• fuse spleen cells with cell line to make hybridomas
• isolate clones
• screen supernatants for desired antibody
• expand positive clones

Question 3

4 elements in the back bone for all ABO antigens

Answer 3

2 galactose
fructose
N-acetyl-galactosamine

Question 4

extra sugar for type A

Answer 4

N-acetyl- galactosamine

Question 5

extra sugar for type B

Answer 5

galactose

Question 6

Anti-ABO antibody type, implications

Answer 6

IgM, means they will not cross placenta

Question 7

acute intravascular hemolytic transfusion run is mediated by:

Answer 7

complement (classical pathway)

Question 8

Rh antibodies type and implications

Answer 8

IgG, can cross placenta

Question 9

testing for Rh antibodies

Answer 9

coombs test

Question 10

direct coombs test uses ___from patient

Answer 10

RBCs

Question 11

indirect coombs test uses ___ from patient

Answer 11

plasma

Question 12

what is the “anti-human globulin” used in coombs testing?

Answer 12

mixture of anti-IgG and anti-complement, used to determine if there is IgG or complement present on RBC surface in direct test or in plasma in indirect test

Question 13

direct ELISA tests for presence of-

Answer 13

antigen

Question 14

indirect ELISA tests for presence of-

Answer 14

antibody

Question 15

how can recent infections be confirmed using ELISA?

Answer 15

look at labs from acute and convalescing phases of illness, 4x or greater difference in IgG = acute infection

Question 16

what is the advantage of western blot?

Answer 16

very specific, allows for determination of which viral proteins are reacting with antibody

Question 17

2 types of “scatter” in flow cytometry

Answer 17

forward = size
side = internal granularity/complexity

Question 18

what is antibody staining in flow cytometry?

Answer 18

use antibodies that have been conjugated with fluorophores to detect cellular antigens (FACS)

Question 19

why is “threshold” important in flow cytometry?

Answer 19

focuses the data on the cells of interest

Question 20

2 mitogens

Answer 20

PHA- phytohemoagglutinin

PWM- pokeweek

Question 21

3 substances that can be used to determine the extend of cell proliferation

Answer 21

BrdU
titrated thymidine
CFSE

Question 22

how to interpret results when cells have been treated with CFSE?

Answer 22

if cells are actively proliferating, will lose fluorescence (diluted)

Question 23

left shift indicates

Answer 23

increase in neutrophils