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.
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.