Ab-Ag Interactions Flashcards Preview

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Flashcards in Ab-Ag Interactions Deck (23):
1

Ab AVIDITY

Strength of multivalent antiserum to multivalent Ag
Most Ags have numerous EPITOPES and are also mulltivalent --> Multiple Ab species that can bind to several of these epitopes
NOT TESTED ON THIS WORD

2

Absorption

The use of cross reacting material to remove the activity that causes the cross reactivity

3

Affinity Chromatography

Used to purify many immunological reagents

4

Monoclonal Ab

A single clone of one B-cell is produced by fusing that cell to a tumor cell and isolating the clone with the Ab specificity of interest
NO HETEROGENEITY
Have low affinity, but no avidity

5

Steps to making a monoclonal Ab

Immunize an animal
Isolate spleen cells from immunized animal
Fuse spleen cells to cancer cells
Select only those that are hybrids
Clone the hybrids
Select clone with specificity that you are interested in

6

Problem with using non human Abs in humans

Have an immune response to non-human Abs

7

Chimeric

Variable regions from mouse
Constant from humans

8

Humanized

Only the points of contact with Ag remain mouse

9

Human monoclonal

Made through molecular biology techniques

10

Immunoassays - ELISA

-Ag stuck to bottome of well
-Ab is added, allowed to incubate, excess Ab washed
-2nd Ab that has enzyme linked molecule (that binds to 1st Ab) added
-Incubation, and excess 2ndary washed
-2ndary Ab detected by adding a chemical reagent that turns color in presence on En bound to 2ndary Ab

11

Immunofluorescence

Tissue or cells are reacted with antisera specific for a cell marker or pathogen
Unbound antiserum is removed by washing
A 2nd Ab specific for the 1st Ab is added and allowed to bind
Attached to 2nd Ab is a fluorescent molecule
Cells viewed through microscope

12

Fluorescence Activated Cell Sorter - Flow cytometry

Mixture of cells are labeled with fluorescent Abs
Analyze cells with lasers and sort

13

Western Immunoblot

-Mixture of proteins such as cell lysates or serum are separated electrophoretically
-Separated proteins bound to nitrocellulose paper
-Ab to protein of interest then added, wash unbound, add 2nd Ab
-Ab detected by En-linked 2ndary Ab

14

Possible constant light chain regions

Kappa and lambda

15

Epitope

Part bound by Ab

16

Types of Epitopes

Linear
Discontinuous

17

What is meant by Abs are HETEROGENOUS

Difference in how and where Abs bind to same Ag

18

Polyclonal Abs

Abs made from diff B-cells for same Ag

19

Uses of Abs in Lab

-Abs are used to measure many biological parameters

20

How to make Abs more specific

-Affinity purify
-Absorb non-specific antisera
-Monoclonal Ab

21

Affinity Purify Ab

Make Ab more specific by purifying the Ab you want
-Add Ag with beads
-Add diff Abs
-Wash off Abs not attached to Ag and bead
-Remove bead from Ab
-Left with specific Ab

22

Absorb Cross Reacting Ab

Remove the contaminated Ab
-Pour mixture of Ab into Ag attached to beads (that arent interested in)
-Pull out Abs not attached
-Pulled out purified Ab

23

Serum Sickness

Immune reaction to Ab (usually not our own)
-We are sometimes given Abs (that are foreign) to help, but build up Abs against foreign Ab
-Next time injected will have serum sickness