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microbio > lecture 14 chapter 18 > Flashcards

lecture 14 chapter 18 Flashcards

1
Q

the use of antibody molecules to “report” or signal the presence the molecule, cell, or entity to which they bind

A

immunoassays

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2
Q

examples of nature and properties of antibody molecules

A

binding specificity
binding strength
ease of purification
multiple classes
ease of production
east to modify

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3
Q

serum from immunized individuals or experimental animals was the source of the antibodies, but was soon replaced by _________, then by ________

A

bulk purified igG, affinity purified antibody molecules

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4
Q

the study of in vitro antibody-antigen interactions

A

serology

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5
Q

individual not yet exposed to antigen

A

seronegative

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6
Q

_______ has no specific antibodies to that pathogen

A

seronegative

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7
Q

individual has been exposed to antigen

A

seropositive

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8
Q

_______ has produced specific antibodies to pathogen

A

seropositive

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9
Q

any in vitro test that uses antibody-antigen interactions to detect or quantify given antigens or antibodies in a sample

A

immunoassay

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10
Q

simple, but common examples of immunoassays

A

binding of known antibodies identifies unknown pathogen

binding of patient’s antibodies to known pathogen demonstrates current or previous infection

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11
Q

antibody concentration determined by _______

A

serial dilution

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12
Q

serial dilutions are

A

sequential 2- or 10- fold dilutions

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13
Q

in serial dilutions, antigens are added to each _______

A

dilution

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14
Q

_______ is reciprocal of last dilution that gives a detectable antigen-antibody reaction

A

titer (concentration)

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15
Q

example of antibody titer

A

positive observed in 1:256 but not 1:512, antibody titer is 256

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16
Q

where are serial dilutions usually done?

A

in plastic microtiter platers

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17
Q

________ bind yo various epitopes on antigen

A

polyclonal antibodies

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18
Q

what happens with polyclonal antibodies?

A

animals immunized with whole or partial agent; resulting antibodies are collected from animal’s serum

multiple B cells respond, produce mix of antibodies to variety of epitopes

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19
Q

in polyclonal antibodies, more complex antigens will yield more _______

A

antibodies

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20
Q

some polyclonal antibodies may bind ______

A

closely related organisms (for example, Shigella sp. and E. coli outer membrane proteins)

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21
Q

________ produced by animals immunized with IgG from human serum

A

anti-human igG antibodies

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22
Q

anti-human antibodies are available ________

A

commercially

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23
Q

example of anti-human IgG antibody

A

mouse, anti-human IgG (antibody raised in a mouse that was immunized with human IgG)

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24
Q

antibodies such as anti-human IgG antibodies are _______ for immunoassays

A

invaluable

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25
Q

_______ recognize a single epitope; produce only in the laboratory (not naturally occurring)

A

monoclonal antibodies (MAbs, Mabs, mAbs)

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26
Q

monoclonal antibodies require

A

considerable “up-front” investment of time/resources

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27
Q

the production of monoclonal antibodies is a complex process that requires

A

fusing animal B cells and cells that divide continuously in culture

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28
Q

monoclonal antibodies produce

A

clones of a single cell and antibody to one epitope

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29
Q

Mabs can be used in

A

immunoassays or in therapy

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30
Q

Mabs medications end in

A

“-mab”

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31
Q

______ neutralized anthrax toxin

A

obiltoxaximab

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32
Q

______ can be tagged with drug or toxin and delivered to specific cells in vivo

A

Mabs

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33
Q

_______ are being developed against certain types of cancers

A

antibody-drug conjugates (ADCs)

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34
Q

example of antibody-drug conjugates

A

immuno-toxin therapeutics

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35
Q

the normal humoral response is, by nature _____ because numerous clones of B-cells will collectively produce antibodies that react with numerous epitopes

A

polyclonal

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36
Q

each individual B-cell (plasma cell) produces antibody to _______

A

only ONE epitope

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37
Q

if we could isolate a PARTICULAR plasma cell producing against an epitope of OUR choosing, we will have a plasma cell making a _______ antibody

A

monoclonal

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38
Q

plasma cells isolated from animals (humans) do not grow in ______

A

culture

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39
Q

cells die within a few days of removal

A

plasma cells isolated from animals

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40
Q

how do you find the plasma cell producing the antibody you want from billions of individual plasma cells?

A

fusing the plasma cells with an “immortal” myeloid cell line allows hybrid cell to live and grow in a culture (a so-called hybridoma)

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41
Q

fused plasma cells (hybridoma) continues to produce _______

A

antibody forever

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42
Q

treating fused plasma cells with a drug that kills non-fused myeloid cells does what?

A

selects for survival of only hybridoma (fused) cells

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43
Q

diluting the fused cell population prior to plating such that each microtiter well has only one cell will allow for growth of

A

a “pure” hybridoma cell line

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44
Q

_________ can be “humanized”

A

monoclonal antibodies

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45
Q

what does being “humanized” mean?

A

most of the animal-derived antibody molecule is replaced with human equivalent using recombinant DNA techniques

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46
Q

what is a humanized monoclonal antibody called?

A

a rhuMab: recombinant humanized monoclonal antibody

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47
Q

rhuMabs have ______ half-life than standard monoclonal antibodies

A

longer

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48
Q

human immune system less likely to destroy _____ antibodies

A

rhuMab

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49
Q

rhuMab medications have named ending in

A

“-zumab”

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50
Q

some rhuMabs are used to treat

A

certain cancers

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51
Q

_________ won a Nobel prize in 1984 for their work on monoclonal antibodies

A

Kohler and Milstein

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52
Q

Adalimumab drug name

A

Humira

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53
Q

Bezlotoxumab drug name

A

Zinplava

54
Q

Capromab pendetide drug name

A

Prostascint

55
Q

Cedelizumab drug name

A

CIMZIA

56
Q

IMC-225 drug name

A

Erbitux

57
Q

Cosfroviximab drug name

A

ZMapp

58
Q

Adalimumab treatment

A

rheumatoid arthritis

59
Q

Adalimumab targets

A

TNF alpha

60
Q

Adecatummab MT-201 treatment

A

breast cancer
other neoplasias

61
Q

Adecatummab MT-201 targets

A

EpCAM (CD326) epithelial cell adhesion molecule

62
Q

Capromab pendetide treatment

A

prostatic carcinoma cells detection

63
Q

Capromab pendetide targets

A

tumor surface antigen PSMA

64
Q

Bexlotoxumab targets

A

clostridium difficile colitis anti-B toxin

65
Q

Bexlotoxumab treatment

A

pseudomembranous colitis

66
Q

removab targets

A

EpCAM, CD3

67
Q

removab treats

A

ovarian cancer

68
Q

cedelizumab treatment

A

prevent organ transplant rejection

69
Q

cedelizumab targets

A

CD4

70
Q

IMC-225 treats

A

metastaticolorectal cancer

71
Q

IMC-225 targets

A

EGFR

72
Q

cosfroviximab treats

A

disease modifying Ebola virus

73
Q

cosfroviximab targets

A

Ebola virus glycoprotein

74
Q

diridavumab treats

A

infectious disease, influenza A

75
Q

diridavumab targets

A

influenza A hemagglutinin

76
Q

_______ can detect a given antigen, or can detect antibodies that bind to a given antigen

A

labeled antibodies

77
Q

_______ identify unknown antigen

A

direct immunoassays

78
Q

in direct immunoassays, where are antigens attached?

A

to solid surface

79
Q

in direct immunoassays, _____ of known specificity added-washed free of unbound

A

labeled antibodies

80
Q

specific binding identifies ______ in direct immunoassays

A

antigen

81
Q

______ detect specific antibodies

A

indirect immunoassays

82
Q

in indirect immunoassays, ______ attached to surface; serum added

A

known antigen

83
Q

in indirect immunoassays, _____ removed unbound primary antibodies

A

washing

84
Q

_______ are added after washing in indirect immunoassays

A

secondary antibodies (labeled anti-human IgG)

85
Q

in indirect immunoassays, washing removes unbound: binding identifies ________

A

primary antibody bound to antigen

86
Q

direct immunoassays identify

A

unknown antigen

87
Q

indirect immunoassays detect

A

specific antibody in patient’s serum

88
Q

fluorescent antibody (FA) test

A

use fluorescence microscopy to locate labeled antibodies bound to antigens fixed to microscope slide

89
Q

in the fluorescent antibody test, the size/shape of antigens are seen, but

A

procedure is time-consuming

90
Q

in the fluorescent antibody test, using different colored labels can

A

detect more than one antigen

91
Q

in fluorescent antibody test, rabies can be detected in

A

brain tissue

92
Q

enzyme-linked immunosorbent assay is also called

A

ELISA

93
Q

direct ELISA is also known as the

A

sandwich method

94
Q

steps of direct ELISA

A

antigen “captured” by antibodies attached to surface

enzyme-labeled antibody added to detect antigens

95
Q

examples of direct ELISA

A

rapid group A strep tests, home pregnancy kits

96
Q

indirect ELISA detects antibodies in ______

A

human blood and serum

97
Q

blood is routinely screened for antibodies to _____ in indirect ELISA

A

HIV

98
Q

indirect ELISA may yield false positives; more reliable test such as _____ used to confirm

A

western blotting

99
Q

ELISA is a form of the general term ______, which is synonymously with ELISA

A

enzyme immunoassay (EIA)

100
Q

in ELISA, antibodies are labeled with

A

enzyme (such as peroxidase)

101
Q

in ELISA, enzyme converts colorless substrate to colored product; detected using ______

A

colorimetric assay

102
Q

ELISA is relatively easy and the basis of many

A

rapid tests, blood screenings

103
Q

ELISA is often done in ______ to test many samples at one

A

microtiter plate

104
Q

pregnancy tests test for the presence of ____ in urine

A

hCG

105
Q

the covid antigen test tests for the presence of

A

SARS-CoV2 spike antigen

106
Q

_______ was developed as a method to detect the presence of a specific protein in a complex mixture such as a cell lysate

A

western blotting

107
Q

in western blotting, proteins are first separated by size using

A

SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis)

108
Q

steps of western blotting

A

proteins are first separated by size

serum (or purified antibody) is added; unbound antibodies washed off

enzyme- or radioactively labeled secondary antibodies added; unbound washed off

label is detected (indicates which proteins on the blot were recognized by antibodies in the serum)

109
Q

_______ counts, sorts cells labeled with fluorescent antibodies

A

fluorescence-activated cell sorter (FACS)

110
Q

for examples, fluorescence-activated cell sorter can track progression of HIV by determining

A

serum levels of CD4 T cells

111
Q

western blots are used extensively

A

in research laboratories

112
Q

in which test does each specimen need to be examined individually?

A

fluorescent antibody (FA) tests

113
Q

which test is widely used to diagnose and monitor the progression of diseases involving white blood cells?

A

fluorescence-activated cell sorter

114
Q

which test is used to characterize different types and stages of lymphomas and leukemias, and to diagnose immunodeficiency diseases

A

fluorescence-activated cell sorter

115
Q

which test is used in clinical labs to diagnose certain diseases including syphilis, rabies, and others

A

fluorescent antibody (FA) tests

116
Q

which test is used in pregnancy tests, strep throat, Lyme disease, West Nile fever, hepatitis B and C, HIV disease

A

enzyme-linked immunosorbent assay (ELISA)

117
Q

immunoassays may involve visible _______

A

antigen-antibody aggregates

118
Q

antibodies that cross-link antigens create larger “mouthfuls” for

A

phagocytic cells

119
Q

clumped antigen-antibody complexes form the basis of

A

agglutination reactions and precipitation reactions

120
Q

_______ cross-link relatively large particles such as cells

A

agglutination reactions

121
Q

_____ used for blood typing

A

hemagglutination

122
Q

________ antibody mixed with antigen (cells, microorganisms); binding yields clumping

A

direct agglutination test (DAT)

123
Q

______ used when antigens are small

A

passive agglutination

124
Q

_______ attached to particles (latex beads) to form larger aggregates

A

antibodies or antigens

125
Q

latex beads in agglutination reactions are mixed with

A

drop of body fluid or microbial culture

126
Q

antibodies bound to soluble antigens can cross-link, form

A

precipitate

127
Q

in precipitation reactions, there are certain relative concentrations of

A

antibody and antigens

128
Q

proper concentrations of antibody and antigen in precipitation reactions are determined by

A

placing separate suspensions near each other in gel, allow molecules to diffuse

129
Q

precipitate will form in

A

“zone of optimal proportions”

130
Q

although largely replaced, _____ illustrates immunodiffusion test

A

ouchterlony technique

microbio (8 decks)

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