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Monoclonal antibodies against human antithrombin III
Z Hrkal1, H Cajthamlová, J T Novák
1Institute of Hematology and Transfusion, Praha, Czechoslovakia.
Hybridoma
|October 1, 1991
Summary
Researchers developed three monoclonal antibodies (D8, B11, C5) targeting human antithrombin III (AT). Antibody D8 showed the highest affinity for AT, while D8 and B11 recognized AT in different biological samples and complexes.
Area of Science:
- Immunology
- Biochemistry
- Protein Science
Background:
- Human antithrombin III (AT) is a critical regulator of coagulation.
- Monoclonal antibodies are valuable tools for studying protein interactions and functions.
- Characterizing antibody specificities is essential for diagnostic and therapeutic applications.
Purpose of the Study:
- To produce and characterize monoclonal antibodies against human antithrombin III (AT).
- To determine the binding affinities of these antibodies to AT.
- To investigate the reactivity of antibodies with AT in various biological contexts.
Main Methods:
- Production of three monoclonal antibodies: D8, B11, and C5.
- Determination of apparent dissociation constants (Kd app) using ELISA.
- Analysis of antibody reactivity with isolated AT and plasma/serum samples via immunoblotting with native PAGE.
Main Results:
- Antibodies D8, B11, and C5 demonstrated varying specificities against human AT.
- Apparent dissociation constants (Kd app) were determined: D8 (2.4 nmole), B11 (13 nmole), C5 (24 nmole), indicating differential affinities.
- All antibodies recognized isolated AT on immunoblots. D8 reacted with plasma and serum AT, while B11 recognized thrombin-antithrombin (TAT) complexes in serum.
Conclusions:
- The generated monoclonal antibodies (D8, B11, C5) are specific tools for human AT research.
- Antibody D8 exhibits high affinity and broad reactivity with AT in different forms.
- Antibody B11 shows utility in detecting TAT complexes, relevant for coagulation studies.