A novel anti-aldolase C antibody specifically interacts with residues 85-102 of the protein

Simona Langellotti, Maurizio Romano, Corrado Guarnaccia, Vincenzo Granata, Stefania Orrù, Adriana Zagari, Francisco E. Baralle, Francesco Salvatore

Research output: Contribution to journalArticle

Abstract

Aldolase C is a brain-specific glycolytic isozyme whose complete repertoire of functions are obscure. This lack of knowledge can be addressed using molecular tools that discriminate the protein from the homologous, ubiquitous paralog aldolase A. The anti-aldolase C antibodies currently available are polyclonal and not highly specific. We obtained the novel monoclonal antibody 9F against human aldolase C, characterized its isoform specificity and tested its performance. First, we investigated the specificity of 9F for aldolase C. Then, using bioinformatic tools coupled to molecular cloning and chemical synthesis approaches, we produced truncated human aldolase C fragments, and assessed 9F binding to these fragments by western blot and ELISA assays. This strategy revealed that residues 85-102 harbor the epitopecontaining region recognized by 9F. The efficiency of 9F was demonstrated also for immunoprecipitation assays. Finally, surface plasmon resonance revealed that the protein has a high affinity toward the epitope-containing peptide. Taken together, our findings show that epitope recognition is sequence-driven and is independent of the three-dimensional structure. In conclusion, given its specific molecular interaction, 9F is a novel and powerful tool to investigate aldolase C's functions in the brain.

Original languageEnglish
Pages (from-to)708-717
Number of pages10
JournalmAbs
Volume6
Issue number3
DOIs
Publication statusPublished - 2014

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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    Langellotti, S., Romano, M., Guarnaccia, C., Granata, V., Orrù, S., Zagari, A., Baralle, F. E., & Salvatore, F. (2014). A novel anti-aldolase C antibody specifically interacts with residues 85-102 of the protein. mAbs, 6(3), 708-717. https://doi.org/10.4161/mabs.28191