Specific oligobodies against ERK-2 that recognize both the native and the denatured state of the protein.
Oligonucleotide aptamer(s), obtained by using the SELEX procedure, has been used as reagents to recognize different molecules with high affinity and specificity. However, until recently, it was not possible to obtain oligonucleotide-based reagents able to recognize proteins with high specificity in assays typical of antibodies, such as immunohistochemistry, Western blotting and immunoprecipitations. Here, we show the results obtained by applying the strategy of "target switching" to obtain specific polyclonal and monoclonal oligobodies against the protein ERK2. We were able to develop highly specific polyclonal oligobodies by using only one selection step with a temporary target and one selection step with the final target (ERK2). Since only two selection steps were required, these results demonstrate that it is possible to obtain specific reagents against a protein without a need for an "in vitro evolution" using many selection steps, or error-prone polymerases. After one additional selection step, the polyclonal oligobodies were cloned to obtain a highly specific monoclonal oligobody.
Bianchini
M
M
Instituto de Investigaciones Bioquímicas-Fundación Campomar, Patricias Argentinas 435, 1405, Buenos Aires, Argentina.
Radrizzani
M
M
Brocardo
M G
MG
Reyes
G B
GB
Gonzalez Solveyra
C
C
Santa-Coloma
T A
TA
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D004277:DNA, Single-Stranded / Q000276:immunology; D006801:Humans; D019950:Mitogen-Activated Protein Kinase 1 / Q000032:analysis* / Q000276:immunology; D008969:Molecular Sequence Data; D011489:Protein Denaturation; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D004277:DNA, Single-Stranded; D019950:Mitogen-Activated Protein Kinase 1