scholarly journals Importance of high-throughput cell separation technologies for genomics/proteomics-based clinical diagnostics

2002 ◽  
Author(s):  
James F. Leary ◽  
Peter Szaniszlo ◽  
Tarl W. Prow ◽  
Lisa M. Reece ◽  
Nan Wang ◽  
...  
Keyword(s):  
High Throughput ◽  
Cell Separation ◽  
Clinical Diagnostics

Clog-free cell filtration using resettable cell traps

Lab on a Chip ◽  
2014 ◽  
Vol 14 (15) ◽  
pp. 2657-2665 ◽  
Author(s):  
William Beattie ◽  
Xi Qin ◽  
Lin Wang ◽  
Hongshen Ma
Keyword(s):  
High Throughput ◽  
Cell Separation ◽  
Free Cell ◽  
Separation Mechanism ◽  
Cell Filtration ◽  
Capture And Release

A microfluidic cell separation mechanism created using constrictions with adjustable size that can selectively capture and release cells, thereby enabling high throughput size and deformability based cell separation without clogging.

scholarly journals Directed Evolution of Aptamer Discovery Technologies

2021 ◽  
Author(s):  
Diana Wu ◽  
Chelsea Gordon ◽  
John Shin ◽  
Michael Eisenstein ◽  
Hyongsok Tom Soh
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Click Reaction ◽  
Clinical Diagnostics ◽  
High Quality ◽  
Affinity Reagents ◽  
Wide Range ◽  
Target Binding ◽  
Modified Nucleobases ◽  
Parallel Fashion

Although antibodies are a powerful tool for molecular biology and clinical diagnostics, there are many emerging applications for which nucleic acid-based aptamers can be advantageous. However, generating high-quality aptamers with sufficient affinity and specificity for biomedical applications is a challenging feat for most research laboratories. In this Account, we describe four techniques developed in our lab to accelerate the discovery of high quality aptamer reagents that can achieve robust binding even for challenging molecular targets. The first method is particle display, in which we convert solution-phase aptamers into aptamer particles that can be screened via fluorescence-activated cell sorting (FACS) to quantitatively isolate individual aptamer particles based on their affinity. This enables the efficient isolation of high-affinity aptamers in fewer selection rounds than conventional methods, thereby minimizing selection biases and reducing the emergence of artifacts in the final aptamer pool. We subsequently developed the multi-parametric particle display (MPPD) method, which employs two-color FACS to isolate aptamer particles based on both affinity and specificity, yielding aptamers that exhibit excellent target binding even in complex matrices like serum. The third method is a click chemistry-based particle display (click-PD) that enables the generation and high-throughput screening of non-nattural aptamers with a wide range of base modifications. We have shown that these base-modified aptamers can achieve robust affinity and specificity for targets that have proven challenging or inaccessible with natural nucleotide-based aptamer libraries. Lastly, we describe the non-natural aptamer array (N2A2) platform, in which a modified benchtop sequencing instrument is used to characterize base-modified aptamers in a massively parallel fashion, enabling the efficient identification of molecules with excellent affinity and specificity for their targets. This system first generates aptamer clusters on the flow-cell surface that incorporate alkyne-modified nucleobases, and then performs a click reaction to couple those nucleobases to an azide-modified chemical moiety. This yields a sequence-defined array of tens of millions of base-modified sequences, which can then be characterized in a high-throughput fashion. Collectively, we believe that these advancements are helping to make aptamer technology more accessible, efficient, and robust, thereby enabling the use of these affinity reagents for a wider range of molecular recognition and detection-based applications.

Abstract 3480: High-throughput size and deformability-based cancer cell separation

2014 ◽  
Author(s):  
Gordon Yip ◽  
Daniel Ionescu ◽  
Edwin Johnson ◽  
Mikael Dick ◽  
Zecong Fang ◽  
...  
Keyword(s):  
Cancer Cell ◽  
High Throughput ◽  
Cell Separation

scholarly journals CNV Workshop: an integrated platform for high-throughput copy number variation discovery and clinical diagnostics

2010 ◽  
Vol 11 (1) ◽  
pp. 74 ◽  
Author(s):  
Xiaowu Gai ◽  
Juan C Perin ◽  
Kevin Murphy ◽  
Ryan O'Hara ◽  
Monica D'arcy ◽  
...  
Keyword(s):  
Copy Number Variation ◽  
High Throughput ◽  
Copy Number ◽  
Clinical Diagnostics ◽  
Integrated Platform ◽  
Number Variation

Pinched flow coupled shear-modulated inertial microfluidics for high-throughput rare blood cell separation

Lab on a Chip ◽  
2011 ◽  
Vol 11 (11) ◽  
pp. 1870 ◽  
Author(s):  
Ali Asgar S. Bhagat ◽  
Han Wei Hou ◽  
Leon D. Li ◽  
Chwee Teck Lim ◽  
Jongyoon Han
Keyword(s):  
Blood Cell ◽  
High Throughput ◽  
Cell Separation ◽  
Inertial Microfluidics

scholarly journals Diagnostic high-throughput sequencing of 2,390 patients with bleeding, thrombotic and platelet disorders

2018 ◽  
Author(s):  
Kate Downes ◽  
Karyn Megy ◽  
Daniel Duarte ◽  
Minka Vries ◽  
Johanna Gebhart ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Sequencing ◽  
Clinical Phenotype ◽  
Careful Consideration ◽  
Clinical Diagnostics ◽  
Genomic Variation ◽  
Panel Test ◽  
Platelet Disorders ◽  
Normal Hemostasis ◽  
And Function

A targeted high-throughput sequencing (HTS) panel test for clinical diagnostics requires careful consideration of the inclusion of appropriate diagnostic-grade genes, the ability to detect multiple types of genomic variation with high levels of analytic sensitivity and reproducibility, and variant interpretation by a multi-disciplinary team (MDT) in the context of the clinical phenotype. We have sequenced 2,390 index patients using the ThromboGenomics HTS panel test of diagnostic-grade genes known to harbour variants associated with rare bleeding, thrombotic or platelet disorders (BPD). The diagnostic rate was determined by the clinical phenotype, with an overall rate of 50.4% for all thrombotic, coagulation, platelet count and function disorder patients and a rate of 6.2% for patients with unexplained bleeding disorders characterized by normal hemostasis test results. The MDT classified 756 unique variants, including copy number and intronic variants, as Pathogenic, Likely Pathogenic or Variants of Uncertain Significance. Almost half (49.7%) of these variants are novel and 41 unique variants were identified in 7 genes recently found to be implicated in BPD. Inspection of canonical hemostasis pathways identified 29 patients with evidence of oligogenic inheritance. A molecular diagnosis has been reported for 897 index patients providing evidence that introducing a HTS genetic test for BPD patients is meeting an important unmet clinical need.

High-throughput rare cell separation from blood samples using steric hindrance and inertial microfluidics

Lab on a Chip ◽  
2014 ◽  
Vol 14 (14) ◽  
pp. 2525-2538 ◽  
Author(s):  
Shaofei Shen ◽  
Chao Ma ◽  
Lei Zhao ◽  
Yaolei Wang ◽  
Jian-Chun Wang ◽  
...  
Keyword(s):  
Microfluidic Device ◽  
High Throughput ◽  
Steric Hindrance ◽  
Cell Separation ◽  
Label Free ◽  
Inertial Microfluidics ◽  
Blood Samples ◽  
Rare Cells

We present a multistage microfluidic device for continuous label-free separation of rare cells using a combination of inertial microfluidics and steric hindrance.

scholarly journals Sheathless High-Throughput Circulating Tumor Cell Separation Using Viscoelastic non-Newtonian Fluid

Micromachines ◽  
2019 ◽  
Vol 10 (7) ◽  
pp. 462 ◽  
Author(s):  
Hyunjung Lim ◽  
Seung Min Back ◽  
Min Ho Hwang ◽  
Dae-Hee Lee ◽  
Hyuk Choi ◽  
...  
Keyword(s):  
High Throughput ◽  
Blood Cells ◽  
Cell Separation ◽  
Separation Efficiency ◽  
White Blood Cells ◽  
Flow Characteristics ◽  
Circulating Tumor Cell ◽  
Size Difference ◽  
Biological Applications ◽  
Mcf 7

Circulating tumor cells (CTCs) have attracted increasing attention as important biomarkers for clinical and biological applications. Several microfluidic approaches have been demonstrated to separate CTCs using immunoaffinity or size difference from other blood cells. This study demonstrates a sheathless, high-throughput separation of CTCs from white blood cells (WBCs) using a viscoelastic fluid. To determine the fluid viscoelasticity and the flow rate for CTC separation, and to validate the device performance, flow characteristics of 6, 13, and 27 μm particles in viscoelastic fluids with various concentrations were estimated at different flow rates. Using 0.2% hyaluronic acid (HA) solution, MCF-7 (Michigan Cancer Foundation-7) cells mimicking CTCs in this study were successfully separated from WBCs at 500 μL/min with a separation efficiency of 94.8%. Small amounts of MCF-7 cells (~5.2%) were found at the center outlet due to the size overlap with WBCs.

scholarly journals Sequencing‐based microsatellite instability testing using as few as six markers for high‐throughput clinical diagnostics

2019 ◽  
Vol 41 (1) ◽  
pp. 332-341 ◽  
Author(s):  
Richard Gallon ◽  
Harsh Sheth ◽  
Christine Hayes ◽  
Lisa Redford ◽  
Ghanim Alhilal ◽  
...  
Keyword(s):  
Microsatellite Instability ◽  
High Throughput ◽  
Clinical Diagnostics
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