10 Immunoassays and Flow Cytometry Flashcards Preview

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Flashcards in 10 Immunoassays and Flow Cytometry Deck (35):
1

Western Blot and Gel based immunoassays are unable to tell what?

Concentration of antibodies in serum

2

What is affinity?

Strength of binding on one site of a molecule on its ligand

3

What is avidity?

Strength of binding of multiple binding sites of a molecule on its ligand

4

What Ig has the highest avidity?

IgM

5

What tests are non-quantitative analysises of antibody levels?

Immunodiffusion and Immunoelectrophoresis gels
Western Blot

6

How do most titers show antibody levels?

By serial dilution

7

What tests are used for blood matching?

Coomb's test

8

What does the presence of RBCs in hemmaglutination test indicate?

Negative antibody presence

9

What does the presence of RBCs in complement fixation assay indicate?

Positive antibody presence, these cells do not get lysed

10

What does a hemagluttination test look for?

Self antibodies

11

What does a complement fication assay look for?

Antibody-antigen complexes, if it binds the RBCs are not lysed

12

No RBC lysis is positive in what test?

Complement

13

What does self-recognizing antibodies to RBCs indicate?

Autoimmune hemolytic anemia

14

Direct Coombs test outline:

Patient blood taken
Antihuman antibodies added
Binding of antibodies means RBCs have antigens recognized by that antibody

15

Indirect Coombs test outline:

Patient Serum obtained
Donor blood added
Antibody binding occurs
If antibody binds, cells clump
Positive means NO TRANSFUSION

16

What is a monospot test for?

Epstein-Barr Virus

17

What test is positive for antibodies if no lysis occurs?

Complement fixation assay

18

How are monoclonal antibodies produced?

Fusion of B cells with Myelonas to make eternal antibody factory called a hybridoma

19

How can we purify antibodies?

Affinity chromatography (Ig binding beads, then wash)

20

How can we make antibodies easy to detect?

Radiolabeling
Enzyme addition
Fluorescent tags

21

What is a primary antibody?

Test sample, may or may not bind to antigen

22

What is a secondary antibody?

Pre-made antibody, known specificity, conjugated to indicator molecule

23

What is serum that is drawn after infection and Ig's have been made?

Convalescent Serum
may be of use in treating another person with the same infection

24

What is serum that is drawn during an active infection?

Acute phase serum

25

What does acute phase serum contain?

High IgM, Low IgG
High Mannose Binding Protein and C reactive protein

26

What does convalescent serum contain?

Antibody specific Ig's

27

What test uses pre-bound known antibodies in a plate of wells?

ELISA

28

What does foward scatter in flow cytometry show?

Cell size

29

What does side scatter in flow cytometry show?

Cell granularity

30

What is a "gated population" in flow cytometry?

A parameter of cell features that can allow you to focus in on a certain group of cell types within a "gate"

31

What allows analysis of two types of cell reactions in flow cytometry?

Double staining by Fluorescence of two different colors

32

What can be used to separate cells?

Fluorescent Activated Cell Sorting in a flow cytometery

33

What color is FITC?

Green

34

What color is PE?

Red

35

Do you ever find monoclonal antibodies in patient serum

(resounding) NO, because of hypermutation