Protein antigens denaturation

In general, the mechanism of heat and alkaline solution for DNA extraction may be based upon a hypothesis, previously proposed for the AR technique.32 Strong alkaline solution may denature and hydrolyze proteins, resulting in breaking cell and nuclear membranes as well as disrupting cross-linkages due to formalin fixation. It is no surprise to observe the similarity between retrieval of nucleic acid and retrieval of protein (antigen) based on a similar chemical reaction of formaldehyde with these two kinds of macromolecules (Fig. 3.1).15"19... 

Refolded protein was used as the capture antigen during hybridoma screening (see Notes 2 and 3). It is also possible to perform a second screen using the denatured protein antigen in order to screen for antibodies that bind masked epitopes within the protein. The resultant MAbs should work for Western blot analysis. 

Denaturation of protein antigens has been extensively studied over the past 70 years and in general has been shown to decrease immunogenicity relative to that of the native form. In addition the antigenic specificity of... 

Samples for assay by Western blotting are separated by electrophoresis in SDS before blotting and antibody staining, so the reacting proteins are denatured. The use of an SDS-denatured antigen to generate the monoclonal antibody response to the "viral coat protein could assure that similar specificities are achieved in the test. 

---