Acoustic impedance-based size-independent isolation of circulating tumour cells from blood using acoustophoresis

Lab on a Chip ◽  
2018 ◽  
Vol 18 (24) ◽  
pp. 3802-3813 ◽  
Author(s):  
S. Karthick ◽  
P. N. Pradeep ◽  
P. Kanchana ◽  
A. K. Sen

Here, we report a label-free method based on acoustic impedance contrast for the isolation of CTCs from peripheral blood mononuclear cells (PBMCs) in a microchannel using acoustophoresis. Applying this method, we demonstrate the label-free isolation of HeLa and MDA-MB-231 cells from PBMCs.

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Olivier Dionne ◽  
François Corbin

AbstractFragile X syndrome (FXS) is the most prevalent inherited cause of intellectual disabilities and autism spectrum disorders. FXS result from the loss of expression of the FMRP protein, an RNA-binding protein that regulates the expression of key synaptic effectors. FXS is also characterized by a wide array of behavioural, cognitive and metabolic impairments. The severity and penetrance of those comorbidities are extremely variable, meaning that a considerable phenotypic heterogeneity is found among fragile X individuals. Unfortunately, clinicians currently have no tools at their disposal to assay a patient prognosis upon diagnosis. Since the absence of FMRP was repeatedly associated with an aberrant protein synthesis, we decided to study the nascent proteome in order to screen for potential proteomic biomarkers of FXS. We used a BONCAT (Biorthogonal Non-canonical Amino Acids Tagging) method coupled to label-free mass spectrometry to purify and quantify nascent proteins of peripheral blood mononuclear cells (PBMCs) from 7 fragile X male patients and 7 age-matched controls. The proteomic analysis identified several proteins which were either up or downregulated in PBMCs from FXS individuals. Eleven of those proteins were considered as potential biomarkers, of which 5 were further validated by Western blot. The gene ontology enrichment analysis highlighted molecular pathways that may contribute to FXS physiopathology. Our results suggest that the nascent proteome of PBMCs is well suited for the discovery of FXS biomarkers.


Antioxidants ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 1112
Author(s):  
Chiara Lanzillotta ◽  
Viviana Greco ◽  
Diletta Valentini ◽  
Alberto Villani ◽  
Valentina Folgiero ◽  
...  

Down syndrome (DS) is the most common chromosomal disorder and the leading genetic cause of intellectual disability in humans, which results from the triplication of chromosome 21. To search for biomarkers for the early detection and exploration of the disease mechanisms, here, we investigated the protein expression signature of peripheral blood mononuclear cells (PBMCs) in DS children compared with healthy donors (HD) by using an in-depth label-free shotgun proteomics approach. Identified proteins are found associated with metabolic pathways, cellular trafficking, DNA structure, stress response, cytoskeleton network, and signaling pathways. The results showed that a well-defined number of dysregulated pathways retain a prominent role in mediating DS pathological features. Further, proteomics results are consistent with published study in DS and provide evidences that increased oxidative stress and the increased induction of stress related response, is a participant in DS pathology. In addition, the expression levels of some key proteins have been validated by Western blot analysis while protein carbonylation, as marker of protein oxidation, was investigated. The results of this study propose that PBMCs from DS children might be in an activated state where endoplasmic reticulum stress and increased production of radical species are one of the primary events contributing to multiple DS pathological features.


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