A novel nano-immunoassay method for quantification of proteins from CD138-purified myeloma cells: biological and clinical utility

Haematologica. 2018 May;103(5):880-889. doi: 10.3324/haematol.2017.181628. Epub 2018 Mar 15.

Abstract

Protein analysis in bone marrow samples from patients with multiple myeloma has been limited by the low concentration of proteins obtained after CD138+ cell selection. A novel approach based on capillary nano-immunoassay could make it possible to quantify dozens of proteins from each myeloma sample in an automated manner. Here we present a method for the accurate and robust quantification of the expression of multiple proteins extracted from CD138-purified multiple myeloma samples frozen in RLT Plus buffer, which is commonly used for nucleic acid preservation and isolation. Additionally, the biological and clinical value of this analysis for a panel of 12 proteins essential to the pathogenesis of multiple myeloma was evaluated in 63 patients with newly diagnosed multiple myeloma. The analysis of the prognostic impact of CRBN/Cereblon and IKZF1/Ikaros mRNA/protein showed that only the protein levels were able to predict progression-free survival of patients; mRNA levels were not associated with prognosis. Interestingly, high levels of Cereblon and Ikaros proteins were associated with longer progression-free survival only in patients who received immunomodulatory drugs and not in those treated with other drugs. In conclusion, the capillary nano-immunoassay platform provides a novel opportunity for automated quantification of the expression of more than 20 proteins in CD138+ primary multiple myeloma samples.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers, Tumor / genetics*
  • Biomarkers, Tumor / metabolism*
  • Humans
  • Immunoassay / methods*
  • Multiple Myeloma / diagnosis
  • Multiple Myeloma / genetics
  • Multiple Myeloma / metabolism*
  • Nanotechnology / methods*
  • Prognosis
  • RNA, Messenger / genetics*
  • Survival Rate
  • Syndecan-1 / metabolism*

Substances

  • Biomarkers, Tumor
  • RNA, Messenger
  • SDC1 protein, human
  • Syndecan-1