Monoclonal antibody binding proteins

Using Antibodies to Probe Structure-Function Relationships in Proteins A monoclonal antibody binds to G-actin but not to F-actin. What does this tell you about the epitope recognized by the antibody ... 

Benzo[a]pyrene (62.5 or 500 pg) was applied once to the shaved backs of male C57BL/6 mice (Bjelogrlic et al. 1994). Mice were killed at different time intervals after the treatment. DNA was isolated from the skin, purified, and analyzed for benzo[a]pyrene-7, 8-diol-9, 10-epoxide-DNA adducts. Skin was also evaluated for monoclonal antibody binding to mouse p53 protein, which has been shown to increase in response to DNA damage. Alterations in p53 are the most frequently observed mutations in human cancer. Benzo[a]pyrene-7,8-diol-9, 10-epoxide-DNA adducts reached their maximum concentration 24 hours after the treatment, and decreased sharply within 1 week, regardless of the dose. An increase in p53 protein was seen only after treatment with 500 pg benzo[a]pyrene. 

Ellgaard, L., Holtet, T.L., Moestrup, S.K., Etzerodt, M. and Thogersen, H.C. (1995). Nested sets of protein fragments and their use in epitope mapping characterization of the epitope for the S4D5 monoclonal antibody binding to receptor associated protein. J. Immunol. Meth. 180, 53-61. 

Jensen, T.H., Jensen, J., Szilvay, A.M. and Kjems, J. (1997). Probing the structure of HIV-1 Rev by protein footprinting of multiple monoclonal antibody binding sites. FEBS Lett. 414, 50-54. 

SJ Kennel. Binding of monoclonal antibody to protein in fluid phase and bound to solid supports. J Immunol Meth 55 1, 1982. 

Low levels of labeling occur because the monoclonal antibody binds an infrequent epitope on a protein or binds with low affinity. 

Wu Y, Simonovsky FI, Ratner BD, Horbett TA. The role of adsorbed fibrinogen in platelet adhesion to polyurethane surfaces a comparison of surface hydrophobicity, protein adsorption, monoclonal antibody binding, and platelet adhesion. J Biomed Mater Res A 2005 74(4) 722-38. 

The capture step involves binding of the target protein to a particular ligand, while contaminants are discarded in the flow-through. For monoclonal antibodies, mostly Protein-A-based chromatography is used, which allows a high degree... 

Immunoaffinity chromatography utilizes the high specificity of antigen—antibody interactions to achieve a separation. The procedure typically involves the binding, to a soHd phase, of a mouse monoclonal antibody which reacts either directly with the protein to be purified or with a closely associated protein which itself binds the product protein. The former approach has been appHed in the preparation of Factor VIII (43) and Factor IX (61) concentrates. The latter method has been used in the preparation of Factor VIII (42) by immobilization of a monoclonal antibody to von WiHebrand factor [109319-16-6] (62), a protein to which Factor VIII binds noncovalenfly. Further purification is necessary downstream of the immunoaffinity step to remove... 

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