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An Example of Antibody
Characterization
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Background:
Molecule A is known to express
on the epithelial cell membrane in human gastric mucosa and some carcinomas. Two
types (mouse and human) of monoclonal antibodies were generated against the
human type molecule A. In order to set up an animal model for preclinical
validation of the molecule, it was important to determine: 1. whether the
antibodies reacted with molecule A in the stomach of cyno monkey, rat and mouse;
2. whether the specificity of the human monoclonal antibody was comparable to
that of the mouse monoclonal antibody. Both antibodies were biotinylated for IHC
analysis. |
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Procedures |
Results |
Picture |
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IHC result analysis |
Mouse antibody (MMab) to molecule A showed
specific cell membrane staining in the gastric mucosa of rat, cyno and human.
Human antibody (HMab) showed similar staining pattern but with non-specific
background. Negative control remained negative |
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IHC with mouse and human antibodies to molecule A
in mouse and rat gastric mucosa |
Mouse antibody to molecule A (left panel) was
negative in mouse stomach (left upper), but showed staining on gastric
epithelial cell membrane of the rat (left down). Human antibody (right panel)
showed similar patterns with non-specific background staining.
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IHC with mouse and human antibodies to molecule A
in cyno monkey and human gastric mucosa |
Mouse antibody to molecule A (left panel) showed
strong membrane staining on both cyno (left upper) and human (left down) gastric
epithelial cells. Human antibody showed similar patterns with non-specific
background staining.
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Conclusion |
Both mouse and human monoclonal antibodies
recognized molecule A from rat, cyno monkey and human. Neither of the antibodies
recognized mouse form of molecule A. The mouse antibody showed specific cell
membrane staining in the gastric mucosa, while, in addition to the membrane
staining, human antibody also reacted with non-specific epitopes in different
types of cells.
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