A novel and highly efficient purification procedure for native human dipeptidyl peptidase 3 from human blood cell lysate

PLoS One. 2019 Aug 7;14(8):e0220866. doi: 10.1371/journal.pone.0220866. eCollection 2019.

Abstract

Dipeptidyl amino-peptidase 3 (DPP3) is an aminopeptidase involved in peptide degradation, including hormone peptides as angiotensin II and enkephalins. DPP3 plasma activity increases in septic patients and correlates with mortality risk. However, the exact physiological role of DPP3 remains unclear and animal studies are necessary to reveal the function of DPP3 in vivo. To this demand, we developed a two-step purification procedure for isolation of native human DPP3 from blood cell lysate (BCL) that is suitable for in vivo applications. With the use of monoclonal antibodies coupled to beads in combination with an ion-exchange chromatography, we recovered 68% of human DPP3 activity from BCL with a purity of ≥ 95%. Purified human DPP3 was assayed for activity and protein concentration using recently published DPP3-activity- and immunoassays. Additionally, protein stability and storage in relevant buffers were tested. Our results provide a promising strategy for fast and efficient isolation of human DPP3. The purified human DPP3 represents the native state of DPP3, suitable for future in vivo applications to investigate the physiological role of DPP3 and its involvement in pathophysiological conditions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal
  • Blood Cells / enzymology*
  • Chromatography, Ion Exchange
  • Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / immunology
  • Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / isolation & purification*
  • Humans
  • Preservation, Biological
  • Protein Stability

Substances

  • Antibodies, Monoclonal
  • Dipeptidyl-Peptidases and Tripeptidyl-Peptidases
  • DPP3 protein, human

Grant support

All authors are employed either at Sphingotec GmbH or Sphingotec Therapeutics GmbH, both companies specialized in research and development of novel biomarkers and their clinical use. There was no dedicated external funding for the present study. Sphingotec Therapeutics GmbH holds patents concerning DPP3 and its diagnostic use in diverse clinical settings as well as interventional strategies. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.