Understanding of key Ab functional concepts Flashcards

1
Q

The interaction between an antibody (Ab) and an antigen (Ag) depends on…

A

four types of non-covalent forces

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2
Q

what are the four types of non covakent forces?

A

1) ionic bonding
2) hydrogen bonding
3) London dispersion
4) Hydrophobic interaction

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3
Q

where are ionic boning found?

A

between oppositely charged residues (amino acids)

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4
Q

where are hydrogen bonding found?

A

the electromagnetic interaction between
hydrogen and two electronegative atoms

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5
Q

where are london dispersion interactions found?

A

between the outer electron clouds
of two or more atoms

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6
Q

where are hydrophobic interactions found?

A

where water forces hydrophobic groups
together

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7
Q

The Ab:Ag complex is formed through

A

weak interactions

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8
Q

Because the Ab:Ag complex is formed through weak
interactions, the complex…

A

readily dissociates

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9
Q

What happens to the Ab:Ag complex in low pH (<3.5)?

A

alters protonation and charge depending on residue pKa

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10
Q

What happens to the Ab:Ag complex in high pH (>10.5) ?

A

alters protonation and charge depending on residue
pKa

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11
Q

High salt concentration (e.g. NaCl > 0.3M) causes

A

ionic displacement

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12
Q

Chaotrophic agents (e.g. cyanates) interfere with

A

H-bonding in water
molecules

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13
Q

Strength of total non-covalent interactions between a single Ag-binding site on an Ab and a single epitope is …

A

…the affinity of the Ab for that epitope

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14
Q

Low affinity Abs bind

A

weakly and tend to dissociate

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15
Q

High affinity Abs bind

A

Ag more tightly and remain associated longer

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16
Q

The interaction of an Ab with and Ag at one site will…

A

…increase the probability of a reaction between Ab and Ag at a different site

17
Q

The strength of such multiple interactions between a multivalent Ab (e.g.
IgM) and multiple epitopes of an Ag is called…

A

… avidity

18
Q

what is valence?

A

The number of epitopes the antibody interacts with- so IgG has a valence of 2, IgM has a valence of 5 (remember rigid structure doesn’t let more than this bind at once). A single Fab fragment has valence of 1.

19
Q

Interactions between Ag-Ab are usually ….

A

… specific

20
Q

Interactions between Ag-Ab are usually specific but what can occur?

A

cross reactivbity

21
Q

Interactions between Ag-Ab are usually specific but cross reactivity can occur with…

A

Ag’s that share an identical epitope or if the Ab specific for
one epitope also binds to an unrelated epitope possessing similar
chemical properties

22
Q

Affinity refers to…

A

… the strength of a signle antibody-antigen interactions. Each IgG antigen binding site typically has high affinity for its target

23
Q

Avidity refers to…

A

the strength of all interactions combined. IgM typically has low affinity antigen binding sites but there are ten of the,, so avidity is high

24
Q

An antibody may react with…

A

… two different epitopes

25
IgG, IgA and IgM can precipitate antigen at...
... equivalemce zone. ALlows for the formation of a solid particle, enabling phagocytosis
26
IgG, IgA and IgM can agglutinate...
.... whole cells at ‘equivalence zone’
27
What is Haemaglutination
agglutination of erythrocytes
28
Antibodies interact with chemically distinct regions of...
... Ag known as epitopes
29
Antibodies that interact with the same Ag may do so through...
... distinct epitopes (improves avidity)
30
In polyclonal antiserum, epitope specific Ab ‘s recognise...
... the same Ag. (the same Ag, different epitopes recognised.)
31
Monoclonal antibodies interact with...
the same epitope on the same Ag
32
Inject rabbit with ...
...human Ag of choice (50 - 1000 ug of AgX)
33
Inject rabbit with human Ag of choice (50 1000 ug of AgX) –
primary response
34
Inject rabbit with human Ag of choice (50 1000 ug of AgX) –primary response. Repeat after ...
... 30 days – secondary response
35
Harvest...
... serum and purify the AgX specific antibodies
36
Mouse immunised with ...
... AgX -> harvest spleen cells and fused with immortalised myeloma cells (hybridoma) -> expand in cell culture.
37
Screen each ...
... cell clone antibody for AgX reactivity
38