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IMSE LEC > FINALS - IMMUNOASSAY 2 > Flashcards

FINALS - IMMUNOASSAY 2 Flashcards

1
Q

agglutination means there will be a formation of

A

large complexes

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2
Q

What would be the particular antibody involved when
it comes to the agglutination reactions

A

IgM

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3
Q

Many of the immunoassays right now are based on those
principle of __ AND ___ reaction

A

agglutination and precipitation

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4
Q

Immunoassays have been developed to detect either __ or ___, and they vary from easily performed manual tests to highly complex automated assays.

A

antigen or antibody

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5
Q

is the initial force of attraction that exists
between a SINGLE FAB SITE on an antibody molecule and a SINGLE EPITOPE or determinant site on the corresponding
antigen

A

affinity

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6
Q

how antige and antibody binds? through ___

A

non covalent bond

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7
Q

Antibodies are capable of reacting with antigens that
are structurally similar to the original antigen that
induced antibody production.

This is known as ___

A

CROSS-REACTIVITY

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8
Q

The strength of attraction depends on the ___
of antibody for a particular antigen.

A

specificity

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9
Q

represents the sum of all the attractive forces
between an antigen and an antibody.

A

Avidity

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10
Q

This involves the strength with which a multivalent antibody binds a multivalent antigen, and it is a measure of the overall stability of an
antigen–antibody complex

A

avidity

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11
Q

it is the forces that keeps the molecules together. We need to stabilized the antigen-antibody
complex that would be the mechanisms of
the ___.

A

avidity

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12
Q

A high avidity can actually compensate for a __

A

low affinity

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13
Q

Stability of the antigen–antibody complex is essential to detecting the presence of _____, whether it
is antigen or antibody.

A

an unknown

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14
Q

antigen and antibody binding is

A

affinity

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15
Q

satbilization of antigen antibody binding

A

avidity

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16
Q

what is the requirement for the precipitation reaction?

A

equal or proportion strength of affinity and avidity

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17
Q

The precipitation reaction will be depending on the ___ of antigen and antibody

A

relative porportion

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18
Q

precipitation curve is also known as

A

precipitin curve

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19
Q

the optimum precipitation or the precipitin
curve that we have occurs in the ___in
which the number of the multivalence sites of the antigen-antibody are approximately equal

A

zone of
equivalence

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20
Q

in the zone of equivalence, the precipitation reaction may be declined on the
other side of that particular equivalence zone due to the excess of either antigen or antibody

rthat is what we called as

A

PROZONE and POST ZONE

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21
Q

excess antibody

A

prozone

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22
Q

his phenomenon of
Antigen combines with only one or two antibody molecules
and cross linkages are formed that would be the ___.

A

prozone

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23
Q

excess antigens

A

postzone

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24
Q

In this case every
available antibody site is bound in a single antigen and no
cross-links are formed

A

POSTZONE

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25
Q

his phenomenon is not
correlated with a lattice network, so basically there is no
lattice network formed on the ___.

A

postzone

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26
Q

Measurement of precipitation by light scattering.

A

o Turbidimetry
o Nephelometry

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27
Q

Determination of Precipitation via Passive
immunodiffusion techniques.

A

o Radial immunodiffusion
o Ouchterlony double diffusion or Ouchterlony
double immunodiffusion

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28
Q

Determination of Precipitation via
Immunoelectrophoretic technique

A

o Rocket Immunoelectrophoresis
o Immunoelectrophoresis
o Immunofixation electrophoresis
o Countercurrent Immunoelectrophoresis

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29
Q

is a measure of the turbidity or cloudiness of a
solution.

A

TURBIDIMETRY:

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30
Q

On that particular turbidimetry the light
scatter occurs in proportion to the __, __ and ____ of the
molecules present in the solution

A

size, shape and the concentration

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31
Q

So the measurement when it comes to
the turbidimetry can be made using a
___ or ___.

A

spectrophotometer or automated clinical
chemistry analyzer

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32
Q

In turbidimetry, A detection device is placed in ___ with the incident light

A

direct line

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33
Q

Collecting light after it has passed through the
solution.

A

turbidimetry

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34
Q

It thus measures the reduction in light intensity
due to reflection, absorption, or scatter

A

turbidimetry

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35
Q

It is recorded in absorbance units, a measure of
the ratio of incident light to that of transmitted
light.

A

turbidimetry

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36
Q

In turbidimetry

The unscattered light is measured at ___
degrees angle from the incident light.

A

180 degrees

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37
Q

If we are referring with the nephelometry the
intensity of ___ is measured.

A

scattered light

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38
Q

measures the light that is scattered at a particular
angle from the incident beam as it passes through a
suspension.

A

nephelometry

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39
Q

It is also the index of the solution
concentration

A

NEPHELOMETRY:

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40
Q

Nephelometers measure light scatter at angles
ranging from ___ degrees to about ___ degrees

A

10; 90

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41
Q

this would be the
reaction; we are allowing to run essential
completion but large particles tend to fall out of solution and decrease the amount of the scatter.

A

End-point nephelometry

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42
Q

the rate of
scattering increases and it is measured
immediately after the reagent is added

A

Kinetic or rate nephelometry

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43
Q

Many automated instruments utilize this principle
(_____) for the measurement of serum proteins.

A

nephelometry

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44
Q

provides accurate and precise
quantitation of serum proteins, and due to
automation, the cost per test is typically lower
than other methods.

A

Nephelometry

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45
Q

Nephelometry can provide ___

A

quantitation of complement components
quantitation of c reactive components and even clotting factors
quantification of immunoglobulins such
as the IgG, IgA, IgM and IgE and even the kappa and lambda light chain

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46
Q

among the precipitation reaction, it is less expesive

A

NEPHELOMETRY

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47
Q

When it comes to the nephelometry the
light detection device is at angle to the ___

A

incident light

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48
Q

If we are referring with the ___ again this would be the intensity of the light transmitted through a particular
medium wherein the unscattered light is measured at 180-degrees angle from the incident light beam.

A

turbidimetry

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49
Q

If we are referring with the ___ this would be the intensity of that scattered light that would be the factor that being measured on that particular nephelometry

can used an angle for about
10 to 90-degrees.

A

nephelometry

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50
Q

The precipitation of antigen–antibody complexes can also
be determined in a support medium such as a ___.

A

gel

51
Q

In this particular GEL consists of a
high-molecular-weight complex polysaccharide derived from____, are used for this purpose.

A

seaweed, and agarose, a purified
agar

52
Q

In passive
immunodiffusion technique, The reagent that we have here is an
___

A

antibody

53
Q

In passive immunodiffusion technique, Yung pinakagamit na gamit natin dito ay
yung ___

A

agar at agarose

54
Q

why agar at agarose is commonly used in passive immunodiffusion technique?

A

it helps to stabilized the
diffusion process and allow the
visualization of the precipitate bonds

55
Q

The rate of diffusion in passive immunodiffusion is affected by the ___

A

size of the particles,
the temperature,
the gel viscosity,
and the amount of hydration.

56
Q

RADIAL IMMUNODIFFUSION (RID): described by

A

JAMES OUDIN

57
Q

was the first to use gels for precipitation
reactions, and he pioneered the technique known as
single diffusion

A

JAMES OUDIN

58
Q

Antibody was incorporated into agarose in a test tube.
The antigen was layered on top, and as the
antigen moved down into the gel, precipitation
occurred and moved down the tube in proportion
to the amount of antigen present.

A

RADIAL IMMUNODIFFUSION (RID):

59
Q

A modification of the single-diffusion technique
was the ___.

A

RADIAL IMMUNODIFFUSION

60
Q

the positive result of RADIAL IMMUNODIFFUSION

A

formation of the precipitate bond.

61
Q

two tchnique tomeasure RID

A

mancini method
fahey and mckelvey method

62
Q

one of the method to measure RID that uses end point method

A

mancini

63
Q

a method to measure RID in which the antigen is allowed to diffuse to completion

A

Mancini

64
Q

in mancini method, the determination of IgM is completed for how many hrs?

A

50-72 hrs

65
Q

in mancini method, the determination of IgG is completed for how many hrs?

A

24 hrs

66
Q

the presence of rig formation in mancini method signifies,

A

antigena nd antibody formation

within the diameter is the ANTIGEN CONCENTRATION

67
Q

a method to measure RID in which the method used is kinetic method, using measurements taken before the point of equivalence is reached

A

fahey and mckelvey method

68
Q

the readings for fahey and mckelvey takeshow many hrs

A

reactions is measured after 18 hrs

69
Q

sources of error RID

A

overfilling and underfilling of wells
nicking the side of the wells when filling
spilling sample outside the well
improper incbatio time and temp
incorrect measurement

70
Q

RID has been used to measure ___

A

Ig M, G , A, and complement components

71
Q

it is simple to perform and requires no instrumentation but it is fairly expensive to run

A

Radial immunodiffusion

72
Q

I radial diffusion, we can as well use ELISA but not for __

A

not for low volume analytes such as Ig D or IgG

73
Q

in this technique, both antigen and antibody diffuse independently through a semisolid medium in two dimensions, horizontally and vertically

A

ouchterlony double diffusion

74
Q

wells are cut in a gel - reactants are added to the wells incubate for 12 and 48 hrs in a most chamber - precipitin lines form here the moving front of antigen meets that of antibody

which process is this one?

A

ouchterlony double diffusion

75
Q

ouchterlony double diffusion is also called as

A

ouchterlony double immunodiffusion

76
Q

In letter A in ouchterlony double diffusion

A

common epitope - resembles smooth curve

77
Q

In letter B ouchterlony double diffusion

A

cross line pattern - demonstrate 2 separate reactions
no common epitope
no identity

78
Q

category C ouchterlony double diffusion

A

diffusion of 2 lines - partial identity - with spur?

79
Q

rocket immunoelectrophoresis is also called as

A

laurell technique

80
Q

antibody is distributed i the fell and the antigen is added just like RID

switch ON the electric current

when antigen diffuses, precipitation begis

end result: conical shape precipitin line

A

rocket immunoelectrophoresis

81
Q

why do we need to switch on th electric crrent in rocket imunoelectrophoresis?

A

to facilitate the migration of the antigen

82
Q

end result of the rocket immunophoresis

A

conical shape precipitin line

83
Q

double diffusion technique that incorporates electrophoresis crrent to enhance results

A

immunoelectrophoresis

84
Q

immunoelectrophoresis is introduced by

A

grabar and williams in 1953

85
Q

this is performed as a two major step process and can be used for semi quantitation of a wide range of antigens

A

immunoelectrophoresis

86
Q

2 step in immunoelectrophoresis

A

electrophoresis - separation of protein components

diffusion of antigen and antibodies

87
Q

immunoelectrophoresis takes how many hrs

A

18 - 24 hrs

88
Q

what is the source of antigen of immunoelectrophoresis

A

serum

89
Q

typically the source of antigen is serum, which is electrophoresed to separate out the main protein fractions

a trough is cut in the gell parallel to the line of separation

antiserum is places in the trough, and the gel is incubated for 18 - 24 hrs

these lines or arcs ca be compared in shape , intensity, and location to that of a normal serum to detect abnormalities

A

immunoelectrophoresis

90
Q

used as a screening tool for many serum proteins including the major class of Ig and a qualitative and semi quantitative technique

for detection of myeloma

can detect Waldenstrom macroglobulinemia

A

immunoelectrophoresis

91
Q

in immunoelectrophoresis, we are using goat antibodies and a serum with ___

A

immunodeficiency

92
Q

___, either mataas
yung value nung isang antibody at yung isa naman ay
deficient.

A

B lymphocyte deficiency

93
Q

deficiency that will cause increase bacterial deficiency

A

IgG deficiency

94
Q

in immunodeficiency, what deficency is involved?

A

serum IgA
deficiency.

95
Q

As first described by Alper and Johnson

A

IMMUNOFIXATION ELECTROPHORESIS

96
Q

It is similar to electrophoresis except that it has no trough for the antibody reagent. The antibody is applied directly to the gel surface rather than phase

A

IMMUNOFIXATION ELECTROPHORESIS

97
Q

IMMUNOFIXATION ELECTROPHORESIS reaction time

A

less than 1 hr

98
Q

Modification of immunoelectrophoresis with only 30 - 60 mns of reaction time

A

COUNTERCURRENT IMMUNOELECTROPHORESIS

99
Q

There is a side-to-side distribution of the antigen and
antibody

A

COUNTERCURRENT IMMUNOELECTROPHORESIS

100
Q

negative charge for COUNTERCURRENT IMMUNOELECTROPHORESIS

anode

A

we have antigen 1 (Ag1), antigen 2 (Ag2),
and antigen 3 (Ag3).

101
Q

in countercurrent immunoelectrophoresis,

Kapag sa ___ naman, we have the positive
charge. We have the antibodies there

A

cathode

102
Q

in counter current mmunoelectrophoresis, a side to side means

A

concentration of that Ig or Ag

103
Q

___ are designed for antigens and antibodies that may
be small in size or present in very low concentrations.

A

Labeled immunoassays

104
Q

in labelled immunoassays

the presence of such antigens or antibodies is determined indirectly by using
a ___ to detect whether or not specific binding has taken place

A

labeled reactant

105
Q

in labelled immunoassays
The substance to be measured is known as the __.

A

analyte

106
Q

in labelled immunoassays

__ are bound by molecules that react specifically with them.

A

analytes

107
Q

Characteristics of Labelled Immunoassay

A

▪ Competitive Assay
▪ Non-competitive Assay

108
Q

Classification of Labelled Immunoassay

A

▪ Radioimmunoassay (RIA) -
Competitive and Noncompetitive RIA

▪ Enzyme Immunoassay (EIA) -
Competitive: EIA -
Noncompetitive: Indirect ELISA

▪ Fluorescent Immunoassay (FIA)

109
Q

All the reactants are mixed together simultaneously, and labeled antigen
competes with unlabeled patient antigen for a limited number of antibody
binding sites.

A

Competitive Immunoassay

110
Q

The amount of bound label is Inversely Proportional to the concentration of
the patient antigen.

A

Competitive Immunoassay

111
Q

Antibody, is first passively absorbed to a solid phase (which is also a medium
well).

A

Noncompetitive Immunoassay

112
Q

Unknown patient antigen is then allowed to react with and be captured by the
antibody.

A

non competitive immunoassay

113
Q

After washing to remove unbound antigen, a second antibody with label is
added to the reaction

A

non competitive immunoassay

114
Q

The amount of label measured is Directly Proportional to the amount of
patient antigen

A

non competitive immunoassay

115
Q

what type of antibodies used for labelled immuoassays

A

monoclonal antibodies

116
Q

monoclonal antibodies which are
discovered by __

A

George Kohler and Cesar Milstein

117
Q

. __ remains as a
constant source of high specific antibody.

A

Monoclonal antibodies

118
Q

Aka Calibrators

A

standards

119
Q

Are unlabeled analytes that are made up in known concentrations of the
substances to be measure

A

Standards

120
Q

So the bound and unbound fractions are usually separated by physical means (such
as ___).

A

decantation, centrifugation, and filtration

121
Q

The last step common to all immunoassays is the ___

A

detection of the labeled
analyte

122
Q

Radioimmunoassay Used ___ as a label

A

radioactive substances

123
Q
A

IMSE LEC (15 decks)

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