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Immunosero Lec (Midterm) > 2.1 Labeled immunoassay > Flashcards

2.1 Labeled immunoassay Flashcards

1
Q

Designed for antigens and antibodies that may be small in size or present in very low concentrations

A

Labeled immunoassays

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2
Q

In labelled immunoassay, one of the components of antigen-antibody reaction is usually very low/small in concentration that it cannot be detected in the naked eye

T or F?

A

T

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3
Q

What are the types of labels?

A

Radionuclides
Enzymes
Fluorochromes
Luminescents

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4
Q

Most common types of labels

A

Enzymes

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5
Q

Used in radioactive labels where atoms with unstable niclei that spontaneously emit radiation

cannot be influenced by any other factors

A

Radionuclides

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6
Q

What is used in Beta radiation?

A

3H (Tritium)

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7
Q

Most common radionuclides for diagnosis and research in cellular immunology?

A

Tritium

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8
Q

What is used in Gamma radiation?

A

125I (iodine 125)

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9
Q

Most common in the clinical lab due to its longer half-life and appropriate energy level

A

125I

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10
Q

Gamma radiation is detected using what measuring tool?

A

Crystal Scintillation counter

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11
Q

Radioimmunoasay is developed by?

A

Yalow and Berson (1950s)

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12
Q

This assay is originally used to determined insulin-anti-insulin complexes in diabetic patients

A

Radioimmunoassay

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13
Q

Useful in measuring analytes occuring at trace concentrations

A

Radioimmunoassay

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14
Q

Alternative for RIA?

A

Enzyme labels

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15
Q

react with suitable substrate to produce breakdown products that are either chromogenic, fluorogenic, or luminescent.

A

Enzyme labels

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16
Q

Example of enzyme labels?

A

Horseradish peroxidase
Alkaline phosphatase
B-D-galactosidase

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17
Q

Most common enzyme labels?

A

Horseradish peroxidase

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18
Q

What is the component for the enzyme labels where addition of this causes colorimetric?

A

Substrate

19
Q
  1. Once binding agent (antibody) binds to the antigen, Ab will now become immobilized.
  2. Then we add a new antibody (AHg) so it can attach to the Fc portion of an unknown Ab.
  3. Once attached, it will be immobilized as well.
  4. Since the enzyme molecule is exposed, we will add a substrate for the
    enzyme (either colorimetric or chemiluminescent)

Arrange the order

A

1234

20
Q

Fluorescent labels uses?

A

Fluorochromes / Fluorophores

21
Q

Common fluorescent compounds uses is usually in what form?

A

Isothiocyanate

22
Q

Common fluorescent compounds that has yellow green color?

A

Fluorescein

23
Q

Common fluorescent compounds that has yellow orange color?

A

Auramine Rhodamine

24
Q

Direct immunofluorescence assay uses what sample?

A

Tissues

25
Q

Indirect immunofluorescence assay uses what sample?

A

Serum or plasma

26
Q

Fluorochromes emit light when subjected to?

A

UV radiation

27
Q

What is unknown in Direct immunofluorescence assay?

A

Antigen

28
Q

What is unknown in Indirect immunofluorescence assay?

A

Antibody

29
Q

Uses labels that emit light as the result of electrical or chemical reaction?

A

Chemiluminescence immunoassays

30
Q

The is the chemical reaction to emit light in chemiluminescence?

A

Oxidation

31
Q

Example of luminescent labels?

A

Luminol
Acridinium ester
Ruthenium derivatives
Nitrophenyl oxalates

32
Q

Uses electrochemical compounds that penetrate light when redox reactions occurs

A

Electrochemiluminescence immunoassay

33
Q

Measures the degree of color change of substrate

A

ELISA

34
Q
  • Reactants are mixed simultaneously
  • Labeled antigen competes with unlabelled antigen for a limited number of antibody-binding sites
  • Concentration of labeled antigen is in excess to occupy all antibodybinding sites
  • Label activity is measured to determine unlabeled antigen concentration
    (Inversely proportional)
A

Competitive assay

35
Q
  • Unknown antigen is bound by
    passive adsorption to the solid
    phase
  • Any unbound antigen is
    removed by washing
  • Incubated with the labeled
    antibody
  • Label activity is directly
    proportional with antigen
    concentration
A

Direct noncompetitive assay

36
Q
  • Initial antigen adsorption and
    washing is similar with direct
    method
  • Uses known unlabeled antibody
    to bind the immobilized unknown
    antigen
  • Following washing to remove
    unbound antibodies, known
    labeled antibodies are added
  • Label activity is directly
    proportional with antigen
    concentration
A

Indirect Noncompetitive assay

37
Q

The higher the label activity, the lower the concentration of your unlabeled antigen

Compe or noncompe

A

Compe

38
Q

The amount of labeled activity is directly proportional with the concentration of unknown substance or analyte

Compe or Noncompe

A

noncompe

39
Q

Requires compatible known antibody pair (Unlabeled and labeled) that binds
different epitopes on the unknown antigen

A

Capture immunoassay

40
Q

Uses conjugated antibody that binds with the immobilized
antigen-antibody complex on the plate well

A

Direct capture immunoassay

41
Q

Uses conjugated antibody that binds with the immobilized antigen-antibody complex on the plate well

A

Indirect capture immunoassay

42
Q

Single use, disposable assays in a plastic cartridge that formerly for PCO/home based testing

A

Rapid immunoassay

43
Q

Membrane mateiral used in Rapid immunoassays

A

Nylon

44
Q

Combination of all steps in rapid immunoassay into one

A

Immunochromatography

Immunosero Lec (Midterm) (10 decks)

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