Development of monoclonal oligobodies and chemically synthesized oligobodies

Oligonucleotide aptamers obtained using the SELEX procedure can recognize d ifferent molecules with high affinity. However, for proteins, this recognit ion is limited to native conformations and the specificity has not been cle arly demonstrated by methods such as Western blotting, immunohistochemistry or immunoprecipitations. Using a library of oligonucleotides and a selecti on strategy based on high specificity instead of high affinity, we have rep orted previously the preparation of polyclonal oligobodies, reagents that r ecognize the protein PP2A in a very specific way(4). Here we report a metho d to obtain monoclonal oligobodies. The oligobody developed specifically re cognized both native and denatured states of the protein CPD1 used as a mod el system. We further demonstrate the specificity of the monoclonal oligobo dy using Western blots, immunohistochemistry, and immunoprecipitation, proc edures previously limited only to antibody-based detection. In addition, a confocal microscopy is shown that was obtained using an oligobody made by c hemical synthesis using an oligonucleotide synthesizer, being this the firs t "synthetic antibody" reported.