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Immunology > 6. Antibodies and biotechnology > Flashcards

6. Antibodies and biotechnology Flashcards

1
Q

difference between monoclonal and polyclonal Ab?

A

monoclonal - Ab produced which can only recognise 1 type of Ab
polyclonal - many Ab produced - many epitopes recognised

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2
Q

what cells allow proliferation of cells?

A

tumour/myeloma cells

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3
Q

Advantage of polyclonal Ab

A

good precipitation and agglutination

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4
Q

advantages of monoclonal Ab - 2

A
  1. pure sample - specific binding

2. no batch variation

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5
Q

disadvantages of polyclonal Ab - 3

A
  1. not pure = non-specific binding
  2. limited serum supply = batch variation
  3. ethics - animals
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6
Q

disadvantages of monoclonal Ab -2

A
  1. POOR agglutination and precipitation

2. costly - labour intensive

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7
Q

4 types of blotting techniques - what do each of them detect?

A 
western = proteins 
eastern = carbs/lipids 
Southern = DNA
northern = RNA
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8
Q

what 3 things does western blotting tell us?

A
  1. presence/absence of protein
  2. drug effect
  3. subcellular distribution of proteins, glycosylation and size
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9
Q

method used for extracting pure sample of epitope

A

immunoaffinity production

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10
Q

2 types of immunoaffinity production

A
  1. affinity chromatography

2. immunoprecipitation

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11
Q

3 commonly used proteins for purification in immunoprecipitation

A
  1. 6 x histidine
  2. Gluthione-S-transferase
  3. Maltose binding protein
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12
Q

what does immunohistochemistry detect?

A

distribution of antigens in tissue

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13
Q

what does immunocytochemistry detect?

A

specific ag in whole cells

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14
Q

in IHC - what is used to preserve the cells?

A

paraffin wax

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15
Q

2 uses of ICC in clinical practice?

A

a) cancer diagnosis

b) change in disease with drug

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16
Q

what does ELISA stand for?

A

enzyme-linked immunosorbent assay

17
Q

4 types of ELISA

A
  1. direct
  2. indirect
  3. sandwich
  4. competitive
18
Q

purpose of competitive elisa?

A

test Ag concentration

19
Q

which ELISA start with an Ab coated well?

A

indirect

20
Q

which ELISA only uses 1 AB?

A

direct

21
Q

how to detect outcome in ELISA?

A

add substrate which will react with enzyme on conjugated Ab - changing colour of solution

22
Q

what method is used in diagnosing blood disorders?

A

flow cytometry

23
Q

FACS

A

fluorescence-activated cell sorting

24
Q

in FACS - what does forward scatter determine?

A

size and volume of cell

25
Q

in FACS - what does side scatter determine?

A

granularity, nucleus structure, vesicles inside of cell

26
Q

which cells have large forward and side scatter?

A

monocytes and granulocytes

27
Q

3 problems of lab methods

A
  1. non-specific binding
    2, tissues are auto-fluorescent
  2. Ab need to be optimised
28
Q

solution to non-specific binding

A

use positive and negative controls

29
Q

solution to auto-fluorescent tissues

A

enzymatic markers

30
Q

solution to Ab optimisation

A

alter conc to allow max binding

31
Q

2 enzyme markers

A
  1. horse-radish peroxidase

2. alkaline phosphatase

32
Q

2 fluorochromes

A
  1. fluorescein isiothiocyanate

2, Alexa 488

33
Q

how can Ab be altered so that they don’t cause an autoimmune response in humans?

A

changing constant region of Ab to human

34
Q

name 8 diseases Ab can be used as target therapy

A
  1. cancer
  2. transplant rejection
  3. crohn’s disease
  4. multiple sclerosis
  5. inflammatory lesions
  6. bacterial infections
  7. sepsis
  8. viral infections
35
Q

example of Ab treatment used to combat HER2 on cancer cells

A

Herceptin

36
Q

2 mechanisms of Herceptin

A
  1. binds to EGF on tumour cell - stops proliferation

2. binds to Fc domain - NK cell apoptosis

37
Q

4 ways Ab can be used against cancer

A
  1. naked
  2. attach to toxin
  3. attach radioactive substance
  4. attach enzyme which activates drug
38
Q

hormone which is elevated in pregnancy?

A

hCG

Immunology (11 decks)

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