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

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

Author information

1
Cancer Research Center-IBMCC (USAL-CSIC), Salamanca, Spain.
2
Institute of Biomedical Research of Salamanca (IBSAL), Spain.
3
National Medicines Institute, Warsaw, Poland.
4
Hematology Department, Hospital 12 de Octubre, CNIO, Complutense University, CIBERONC, Madrid, Spain.
5
Hospital Universitario de Salamanca, CIBERONC, Spain.
6
Hospital Germans Trias i Pujol, Barcelona, Spain.
7
Hospital Clinic, Barcelona, Spain.
8
Hospital 12 de Octubre, Madrid, Spain.
9
Clínica Universidad de Navarra, CIMA, IDISNA, CIBERONC, Pamplona, Spain.
10
Cancer Research Center-IBMCC (USAL-CSIC), Salamanca, Spain normagu@usal.es.

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.

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