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cell and molecular biology of the immune system > Lecture 12 > Flashcards

Lecture 12 Flashcards

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1
Q

What do antibodies secreted from BCR bind?

A

Native antigens - they are monoclonal

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2
Q

Where are antibodies found?

A

In the blood

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3
Q

What is a serum?

A

Plasma once blood clot is removed

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4
Q

What is an antiserum?

A

Serum from immunised person

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5
Q

What do antiserums not contain?

A

Cells or clotting proteins

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6
Q

What is a standard way of purifying?

A

Chromatography based on their molecular weight

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7
Q

What comes out for the column first?

A

Big things

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8
Q

What are the two types of chromatography used for antibodies?

A

Gel filtration and affinity

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9
Q

What is purified antiserum sometimes known as?

A

Polyclonal antiserum

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10
Q

What is the same for all antibodies?

A

Their MW

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11
Q

How is the filtration column modified?

A

By using beads

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12
Q

What antibodies will be attached to the beads?

A

Antibodies that are specific to the antigen

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13
Q

What happens to the antibodies that don’t attach to the beads?

A

Go through the column

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14
Q

What do patients with multiple myelomas produce?

A

Large amounts of homogenous antibodies

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15
Q

What are hybridomas?

A

Cells that are fused by myeloma cells with b cells

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16
Q

When did the generation of monoclonal antibodies start?

A

1975

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17
Q

What can live in the drug forever?

A

B cells fusions as they have properties of myelomas

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18
Q

What is a monoclonal antibody?

A

Has come from a single hybridoma

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19
Q

What do many methods using antibodies rely on?

A

Rely on labels attached to the antibodies in order to detect them once they are bound to antigens

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20
Q

What should labels not affect?

A

Antibody or antigen binding

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21
Q

What happens if an antibody has bound to an antigen?

A

An enzyme is bought in and can detect the present of an antigen antibody

22
Q

What is the enzyme?

A

HRP or AP

23
Q

What do many methods of antibodies also use?

A

Secondary antibodies to detect the primary antibody binding to its antigen

24
Q

What does using the same antibody that is not labelled with a secondary antibody that is labelled = ?

A

More sensitivity

25
Q

What is an example of a labelled primary antibody?

A

Mouse IgG monoclonal

26
Q

How long does it take to collect rat anti-mouse antiserum?

A

2-6 weeks

27
Q

What would happen if you were to put one antibody into another animals antibody?

A

It would make another antibody

28
Q

What two ways can antibodies be used in?

A

Research and diagnostics

29
Q

What ways can antibodies be used in research? 3 ways

A

Affinity chromatography, immune precipitation and immunofluorescence microscopy

30
Q

What is affinity chromatography?

A

Can purify molecules from complex mixtures and use antibodies to pull out proteins from a mixture

31
Q

What are magnetic bead isolations?

A

Little beads with antibodies that bind to a Magnet which form a column

32
Q

What happens when the magnet turns off?

A

Antibodies will fall off the magnet pulling the proteins with them

33
Q

What happens in immunofluorescence microscopy?

A

Identifying the location of a protein within the cell

34
Q

What happens to the antibodies in fluorescent microscopy?

A

Antibodies light up different parts of the cell depending on where the protein is being expressed

35
Q

What does western blotting allow?

A

Allows you to look at different proteins inside cells

36
Q

What is a very common technique used in both research and diagnostic?

A

ELISA

37
Q

What does ELISA stand for?

A

Enzyme-linked immunosorbent assay

38
Q

What does ELISA detect?

A

Non linear and linear epitopes

39
Q

What can ELISA be used for in respect to antibodies?

A

Can be used to quantitate amount of antigen present

40
Q

What happens in direct ELISA?

A

Blood is put on a plate and a positive signal is received in the well where the antibody is attached to the antigen

41
Q

What is sandwich ELISA?

A

Even more sensitive and uses less of limited sample

42
Q

What is different to sandwich ELISA compared to direct ELISA?

A

Only dented the well being positive if the antigen has been captured

43
Q

What is flow cytometry (FACS)?

A

A technique used for the characterisation of cells based not their light scattering properties

44
Q

What can the properties be?

A

Either natural or induced by pre-incubation of cells with antibodies labelled with dyes

45
Q

How many cells are able to pass through the lazor at a time?

A

2000 per second

46
Q

What are the two ways cells can move through the lazor?

A

Side scatter and forward scatter

47
Q

At does forward scatter result from?

A

A cells size

48
Q

What does side scatter result from?

A

A cells granularity

49
Q

What can be obtained if you add antibodies to the cell before they go through the lazor?

A

Expression of various molecules

50
Q

What happened in 1975?

A

Never ending supply of monoclonal antibodies

cell and molecular biology of the immune system (12 decks)

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