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Force Spectroscopy of Polyclonal and Monoclonal Anti-Bovine Serum Albumin Antibodies - BSA Complexes

100%
Chtcheglova L., Dietler G.
Acta Physica Polonica A
|
2003
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vol. 104
|
issue 3-4
321-326
EN
The specific interactions between bovine serum albumin and poly- or two monoclonal bovine serum albumin antibodies were studied using force spectroscopy mode of atomic force microscopy. The histograms of the unbinding forces for polyclonal bovine serum albumin antibodies are broad at high antibody concentrations (50 or 270 μg/ml) and narrow at low concentrations (10 or 27 μg/ml), while the histograms for monoclonal antibodies peak at well defined unbinding force. The peak unbinding force depends on the type of antibody and the antibody concentration. In this paper we described and characterized the passive adsorption and covalent immobilization of proteins for tip and sample preparation. Force spectroscopy could serve as a useful method for characterization of antigen-antibody interactions for measuring the specificity of an antibody or to assess the purity of a monoclonal antibody solution and to distinguish between different antibodies.
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Study of Probes and Substrates for Low Temperature Atomic Force Microscopy and Biological Applications

81%
Radenovic A., Bystrenova E., Libioulle L., Dietler G.
Acta Physica Polonica A
|
2003
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vol. 104
|
issue 3-4
373-380
EN
The atomic force microscopy in ultrahigh vacuum and at low temperature demonstrated its excellent capability to reach atomic resolution. Nevertheless in the case of biological samples high resolution has been achieved only in very few cases. We demonstrated here the importance of the appropriate choice of probes and substrates in order to image DNA at low temperature with high resolution. We investigated properties of three types of cantilevers and they were studied by scanning electron microscopy as a function of temperature. A large bending of cantilevers, which were coated from both sides, was observed at low temperatures. Therefore uncoated cantilevers are strongly recommended for low temperature applications. Different methods for immobilization of DNA on the substrate are examined at low temperatures. First images of linear DNA on graphite at 82 K under ultrahigh vacuum conditions are presented.
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