A novel nano-immunoassay method for quantification of proteins from CD138 purified myeloma cells: biological and clinical utility
Misiewicz-Krzeminska I (1), Corchete LA (2), Rojas EA (2), Martínez-López J (3), García-Sanz R (4), Oriol A (5), Bladé J (6), Lahuerta JJ (7), San Miguel J (8), Mateos MV (4), Gutiérrez NC (9); Grupo Español de Mieloma/Programa para el Estudio de laTerapeutica enHemopatıas Malignas (GEM/PETHEMA) Cooperative Study Groups.
(1) CIC-IBMCC (USAL-CSIC), IBSAL, Salamanca, Spain. National Medicines Inst, Warsaw, Poland.
(2) CIC-IBMCC (USAL-CSIC), IBSAL, Salamanca, Spain.
(3) Hematology Department, Hospital 12 de Octubre, CNIO, Complutense University, CIBERONC, Madrid, Spain.
(4) Hospital Universitario de Salamanca, IBSAL, Salamanca, Spain.
(5) Hospital Germans Trias i Pujol, Institut Català d'Oncologia.
(6) Hospital Clinic, Barcelona, Spain.
(7) Hospital Universitario 12 de Octubre, Madrid, Spain.
(8) Clínica Universidad de Navarra, (CIMA), IDISNA, CIBERONC, Pamplona, Spain.
(9) Hospital Universitario Salamanca, IBSAL, Salamanca, Spain.
Protein analysis in bone marrow samples from patients with multiple myeloma has been limited by the low protein concentration obtained after CD138+ selection.
A novel approach based on capillary nano-immunoassay could make it possible to automatically quantify dozens of proteins from each myeloma sample.
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 essential proteins in multiple myeloma pathogenesis was evaluated in 63 newly diagnosed multiple myeloma patients.
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 to automatically quantify the expression of more than 20 proteins in CD138+ primary multiple myeloma samples.
CITA DEL ARTÍCULO Haematologica. 2018 Mar 15. pii: haematol.2017.181628. doi: 10.3324/haematol.2017.181628.