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An Example of Antibody
Characterization
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Background:
Molecule A has been known to
express on the epithelial cell membrane of human gastric mucosa. It has also
been shown to express on the cell membrane of certain types of carcinomas. Two
types (mouse and human) of monoclonal antibodies were generated against the
human type molecule A. To explore the therapeutic potential of the human
antibody and the target, it was important to determine: 1. expression pattern of
the molecule among a variety of normal and tumor tissues; 2. cross-reactivity of
the human monoclonal antibody in human tissues. Biotinylated antibodies were
prepared. IHC was performed in tissue arrays containing 135 human normal tissues from 34 sites and 95 tumor
tissues from 22 cancer types. |
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Procedures |
Results |
Picture |
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IHC result analysis |
Example of a chart showing detailed analysis of
the antibody staining patterns across a wide range of normal and tumor tissues.
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IHC with mouse and human antibodies to molecule A
in normal tissues |
Example of the mouse (MMab, left panel) and human
(HMab, right panel) antibody staining in normal tissues. Both antibodies gave
strong membrane staining to the glandular epithelial cells in the stomach and
ileum. Human antibody also showed cytoplasmic and non-specific staining to other
cell types. |
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IHC with mouse and human antibodies to molecule A
in tumor tissues |
In some types of tumor, the mouse antibody (left
panel) showed specific , but heterogeneous membrane staining. The human antibody
(right panel) showed membrane and cytoplasmic staining with non-specific
background.
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IHC with mouse and human antibodies to molecule A
in tumor tissues |
In some other types of tumor, the mouse antibody
(left panel) showed nuclear staining. The human antibody (right panel) showed
predominantly membrane and cytoplasmic staining with non-specific background.
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Conclusion |
•The mouse monoclonal antibody
detected a general elevation of molecule A
in tumors.
The staining was highly
heterogeneous, i.e. the patterns, positive rates and staining intensity of tumor
cells varied greatly among different and/or same types of tumors, or within the
same tumors.
•The human antibody staining was
predominantly cytoplasmic and homogeneous. The antibody showed a low specificity
to its intended target cells on human tissue sections.
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