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Antibodies as Diagnostic TOols Flashcards

(47 cards)

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1
Q

Which part of the antibody can we attach various things?

A

Fc constant region

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2
Q

Name four categories of things that we can attach to an antibody

A

Enzymes
Drugs
Fluorescent probes
Magnetic beads

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3
Q

Give two examples of enzymes attached to antibodies

A

Peroxidase

Alkaline phosphatase

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4
Q

Give two examples of fluorescent probes attached to antibodies

A

Dyes

Beads of different sizes

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5
Q

Give an example of why magnetic beads may be attached to antibodies

A

Purification of cell types

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6
Q

Give an example of a drug attached to antibodies

A

Kadcyla (anti-HER2 linked to emtansine)

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7
Q

Antibodies can be made against things other than cellular proteins. Give two examples of this.

A

anti DNA antibodies

anti-Ab antibodies e.g. RF

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8
Q

Where are the antibodies used in diagnostic tools actually obtained?

A

Produced by patients

Manufactured antibodies

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9
Q

Give two scenarios where a patient would produce antibodies which can be used as diagnostic tools

A

In autoimmune disease

For defence against infection

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10
Q

Give three examples of manufacture antibodies

A

Antisera from immunised animals (contain lots of different antibodies)
Monoclonal antibodies
Genetically engineered antibodies

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11
Q

What do you call it when you use an antibody to detect an antibody

A

Indirect labelling

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12
Q

What is the problem with polyclonal antisera from animals?

A

There’s not enough antibodies

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13
Q

What has replaced antisera from animals?

A

Monoclonal antibodies

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14
Q

Explain how we generate monoclonal antibodies

A

Immune mouse with antigen
Remove spleen cells (as source of Ab producing b-cells)
Fuse this with external myeloma cells which do not produce any antibodies
Produce hybridomas
Select for hybridomas which produce the antibody of interest

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15
Q

Explain the production of antibodies using recombinant DNA technology

A

Isolate a population of genes encoding Ab variable regions.
Construct fusion protein of variable regions with bacteriophage coat protein.
Cloning a random population of variable regions gives rise to a mixture of bacteriophages (phage display library).
Select a phage with a desired V region which binds to specific antigens.

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16
Q

Give four therapeutic uses of manufactured antibodies

A

Prophylactic (against microbes)
Anticancer therapy
Removal of T-cells from bone marrow grafts
Block cytokine activity

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17
Q

Which antibody is used prophylactically against RSV?

A

Synagis (anti-RSV given to babies)

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18
Q

Which antibody is used against migraines?

A

Anti-calcitonin gene-related peptide (anti-CGRP)

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19
Q

Which antibody is used to eliminate T-cells from bone marrow grafts?

20
Q

Name an antibody used in cancer

21
Q

Give three diagnostic uses of manufactured antibodies

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A

To work out blood group
Quantitative immunoassays
Immunodiagnosis

22
Q

Give three things on which can be quantified using manufactured antibodies (quantitative immunoassays)

23
Q

Give four things which can be diagnosed using antibodies

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A

Infectious diseases
Autoimmune diseases
Allergic diseases
Malignancy

24
Q

What can be used to measure quantitative assays?

25
What does ELISA stand for?
Enzyme linked immunosorbent assay
26
Explain how ELISA works
A plastic plate is coated with an antibody. The suspected antigen is applied to the plate and if present, it will stick to the antibody. A second antibody which has an enzyme attached to it is applied. This binds AT A DIFFERENT SITE to the antigen, and changes colour when binding. The colour is proportional to the amount of antigen there is. Therefore it is quantitative.
27
What is a faster method?
Point of care rapid testing
28
Explain how rapid strip tests work
The strip contains antibodies in gold nanoparticles. Further along, there are two lines: the first line contains more of these antibodies, and the second line contains anti-antibody antibodies (anti-Ig antibodies). When the sample is applied, the gold nanoparticles containing Ab should stick to the antigen, and the other side of the antigen should stick to the first line of Ab. Remaining Ab should stick to the second line of the anti-Ig antibodies.
29
What is the point of having the second anti-Ig line in the strip test?
To ensure the test has worked properly (control).
30
What draws the sample from the strip test towards the antibodies?
Capillary action
31
What is the first and second line called?
Test line | Control line
32
How can immune complex deposition be assessed?
Direct immunohistochemistry of antibodies | Detection of cytokines
33
Give three areas of the body where immune complexes may deposit
Glomerulus of kidney Lungs Skin Joints
34
If we are concerned about immunodeficiency, what three things can be screened for?
Serum Ig levels Specific antibodies Lymphocyte subsets
35
What three things can be used to measure serum Ig levels?
Serum electrophoresis ELISA Nephelometry
36
Antibodies produced against which pneumococcal antigen is detected? (for specific antibody testing)
Antibodies against polysaccharide antigens
37
Specific antibodies can be measured against protein antigens. Name two conditions for which this is the case.
Tetanus | Haemphilus
38
What can be used to detect specific antibodies?
ELISA
39
What can be used to detect lymphocyte subsets?
Flow cytometry
40
What would a single band indicate in serum electrophoresis?
Monoclonal expansion/malignancy
41
How does flow cytometry work?
Various cells and mixed with various fluorescently labelled antibodies. This mixture is passed through a tube, and a laser is shined through which detects different colours and therefore different cell types.
42
In which condition is flow cytometry of utmost importance, and why?
HIV - flow cytometry is used to detect CD4+ T-cells as this is predicts short term outlook in patients who have not had ART.
43
In flow cytometry, CD3+ is used to measure what?
T- cells; it's a pan T cell maker
44
In flow cytometry, CD4+ is used to measure what?
T helper cells
45
In flow cytometry, CD8+ is used to measure what?
Cytotoxic T cells
46
In flow cytometry, CD19+ is used to measure what?
B cells
47
In flow cytometry, CD56+ is used to measure what?
NK cells

Diagnostics (7 decks)

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