Development of monoclonal oligobodies and chemically synthesized oligobodies

MartíN Radrizzani, Mariana G. Brocardo, César Gonzalez Solveyra, Michele Bianchini, Gloria B. Reyes, Eduardo G. Cafferata, Guillermo Vilá Ortiz, Tomás A. Santa-Coloma

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Oligonucleotide aptamers obtained using the SELEX procedure can recognize different molecules with high affinity. However, for proteins, this recognition is limited to native conformations and the specificity has not been clearly demonstrated by methods such as Western blotting, immunohistochemistry or immunoprecipitations. Using a library of oligonucleotides and a selection strategy based on high specificity instead of high affinity, we have reported previously the preparation of polyclonal oligobodies, reagents that recognize the protein PP2A in a very specific way4. Here we report a method to obtain monoclonal oligobodies. The oligobody developed specifically recognized both native and denatured states of the protein CPD1 used as a model system. We further demonstrate the specificity of the monoclonal oligobody using Western blots, immunohistochemistry, and immunoprecipitation, procedures previously limited only to antibody-based detection. In addition, a confocal microscopy is shown that was obtained using an oligobody made by chemical synthesis using an oligonucleotide synthesizer, being this the first "synthetic antibody" reported.

LanguageEnglish
Pages55-60
Number of pages6
JournalMedicina
Volume60
Issue numberSUPPL. 2
Publication statusPublished - 2000
Externally publishedYes

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Oligonucleotides
Immunoprecipitation
Western Blotting
Immunohistochemistry
Proteins
Antibodies
Confocal Microscopy

Keywords

  • Monoclonal oligobodies
  • Synthetic oligobodies

Cite this

Radrizzani, M., Brocardo, M. G., Solveyra, C. G., Bianchini, M., Reyes, G. B., Cafferata, E. G., ... Santa-Coloma, T. A. (2000). Development of monoclonal oligobodies and chemically synthesized oligobodies. Medicina, 60(SUPPL. 2), 55-60.
Radrizzani, MartíN ; Brocardo, Mariana G. ; Solveyra, César Gonzalez ; Bianchini, Michele ; Reyes, Gloria B. ; Cafferata, Eduardo G. ; Ortiz, Guillermo Vilá ; Santa-Coloma, Tomás A. / Development of monoclonal oligobodies and chemically synthesized oligobodies. In: Medicina. 2000 ; Vol. 60, No. SUPPL. 2. pp. 55-60.
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Radrizzani, M, Brocardo, MG, Solveyra, CG, Bianchini, M, Reyes, GB, Cafferata, EG, Ortiz, GV & Santa-Coloma, TA 2000, 'Development of monoclonal oligobodies and chemically synthesized oligobodies', Medicina, vol. 60, no. SUPPL. 2, pp. 55-60.

Development of monoclonal oligobodies and chemically synthesized oligobodies. / Radrizzani, MartíN; Brocardo, Mariana G.; Solveyra, César Gonzalez; Bianchini, Michele; Reyes, Gloria B.; Cafferata, Eduardo G.; Ortiz, Guillermo Vilá; Santa-Coloma, Tomás A.

In: Medicina, Vol. 60, No. SUPPL. 2, 2000, p. 55-60.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Development of monoclonal oligobodies and chemically synthesized oligobodies

AU - Radrizzani, MartíN

AU - Brocardo, Mariana G.

AU - Solveyra, César Gonzalez

AU - Bianchini, Michele

AU - Reyes, Gloria B.

AU - Cafferata, Eduardo G.

AU - Ortiz, Guillermo Vilá

AU - Santa-Coloma, Tomás A.

PY - 2000

Y1 - 2000

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AB - Oligonucleotide aptamers obtained using the SELEX procedure can recognize different molecules with high affinity. However, for proteins, this recognition is limited to native conformations and the specificity has not been clearly demonstrated by methods such as Western blotting, immunohistochemistry or immunoprecipitations. Using a library of oligonucleotides and a selection strategy based on high specificity instead of high affinity, we have reported previously the preparation of polyclonal oligobodies, reagents that recognize the protein PP2A in a very specific way4. Here we report a method to obtain monoclonal oligobodies. The oligobody developed specifically recognized both native and denatured states of the protein CPD1 used as a model system. We further demonstrate the specificity of the monoclonal oligobody using Western blots, immunohistochemistry, and immunoprecipitation, procedures previously limited only to antibody-based detection. In addition, a confocal microscopy is shown that was obtained using an oligobody made by chemical synthesis using an oligonucleotide synthesizer, being this the first "synthetic antibody" reported.

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Radrizzani M, Brocardo MG, Solveyra CG, Bianchini M, Reyes GB, Cafferata EG et al. Development of monoclonal oligobodies and chemically synthesized oligobodies. Medicina. 2000;60(SUPPL. 2):55-60.