Page 314 - Color Atlas of Biochemistry

Immune response 305

A. Monoclonal antibodies

Immunized Antigen Hybridoma cells Antibody test:
mouse
Positive
cultures
A B C A B With HAT
1. Immunization Spleen cells C PEG medium
Fusion Primary culture
2. 3.
Myeloma cells

Cell culture

Mouse with
ascites
Subcloning 4. Cloning
5.
C C C
Production of C
monoclonal C C C
6. antibodies
Culture of Selection C C B B A A
a selected of positive C B A
clone clones
Clone
Cell culture

B. Immunoassay
Enzyme:
E
peroxidase
E
Sample Tracer

Antikörper E E
Antibodies

Incubate
Microtiter plate
1. 2.

Absorption
Working range
-6
C 10 -10 -12 M
Read
E absorption Wash
F Substrate: H O
2 2
Concentration Chromogen: C

F C C F E
C E C
C
C F F C
Incubate
4. 3.

C F
Chromogen + H O 2 Peroxidase Dye + H O
2
1.11.1.7
2

Koolman, Color Atlas of Biochemistry, 2nd edition © 2005 Thieme
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