An automatic microfluidic system for rapid screening of cancer stem-like cell-specific aptamers

Weng, Chih‐Huang; Hsieh, I-Shan; Hung, Lien Yu; Lin, Hsin I.; Shiesh, Shu Chu; Chen, Yuh Ling; Lee, Gwo‐Bin

doi:10.1007/s10404-012-1095-3

Cited by 40 publications

(49 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…While these and other advances are promising, much of the microfluidic SELEX work has focused on the selection step of the aptamer isolation process (Ahn et al 2011, 2012; Kim et al 2011; Park et al 2009). Despite a significant field of literature that has demonstrated integration of amplification with a variety of other functions (Zhang and Xing 2007), there are fewer examples of integrated aptamer isolation microchips with both selection and amplifica-tion stages (Hung et al 2014; Weng et al 2013). Some integrated prototypes suffer from a lack of miniaturization (Hybarger et al 2006) or risk cross-contamination by providing inadequate partitioning between reaction products and reactants (Huang et al 2010).…”

Section: Introductionmentioning

confidence: 99%

Isolation of thermally sensitive protein-binding oligonucleotides on a microchip

Hilton

Olsen

Kim

et al. 2015

Microfluid Nanofluid

View full text Add to dashboard Cite

This paper presents a microfluidic chip capable of isolating thermally sensitive protein-binding aptamer candidates. The chip makes use of bead-immobilized target molecules and DNA (deoxyribonucleic acid) sequences to enable a simplified chip design, in which affinity selection and PCR (polymerase chain reaction) amplification of target-binding sequences occur in temperature-controlled microchambers. Using pressure-driven flow, buffer containing single-stranded DNA molecules with randomized sequences is cycled through a series of affinity selection and PCR amplification steps on microbeads. Successive introduction of the sample to each chamber effects a process of competition whereby DNA strands with weak binding strength to target molecules are rejected in favor of strongly binding sequences. Using bead-based PCR, the amplification step was miniaturized and integrated with affinity selection, resulting in significant reductions in process time and reagent use. As a demonstration, temperature-dependent selection and amplification of single-stranded oligonucleotides that bind to human Immuno-globulin E (IgE) was performed in 4 h, a 20-fold reduction in process time as compared to conventional methods that would require approximately a week. Fluorescent binding assays then demonstrated that the desired temperature specificity was imparted to the aptamer candidates within just one round of selection, and within two rounds the aptamer candidates exhibited enhanced affinity toward IgE.

show abstract

Section: Introductionmentioning

confidence: 99%

Isolation of thermally sensitive protein-binding oligonucleotides on a microchip

Hilton

Olsen

Kim

et al. 2015

Microfluid Nanofluid

View full text Add to dashboard Cite

show abstract

“…Other types of negative-pressure-driven microfluidic components can be found in the literature. [27][28][29] The normally closed valve and micromixer were controlled and driven using a vacuum pump connected to an electromagnetic valve (EMV), and fluid transport was achieved using a syringe pump (Legato 180, KD Scientific Inc., Holliston, MA) connected to the waste outlet channel that was driven with a withdrawing force by opening the inlet to atmospheric pressure. Using a personal computer, control of the fluid in the microchannel can be performed automatically.…”

Section: Methodsmentioning

confidence: 99%

A negative-pressure-driven microfluidic chip for the rapid detection of a bladder cancer biomarker in urine using bead-based enzyme-linked immunosorbent assay

et al. 2013

View full text Add to dashboard Cite

This paper describes an integrated microfluidic chip that is capable of rapidly and quantitatively measuring the concentration of a bladder cancer biomarker, apolipoprotein A1, in urine samples. All of the microfluidic components, including the fluid transport system, the micro-valve, and the micro-mixer, were driven by negative pressure, which simplifies the use of the chip and facilitates commercialization. Magnetic beads were used as a solid support for the primary antibody, which captured apolipoprotein A1 in patients' urine. Because of the three-dimensional structure of the magnetic beads, the concentration range of the target that could be detected was as high as 2000 ng ml À1 . Because this concentration is 100 times higher than that quantifiable using a 96-well plate with the same enzyme-linked immunosorbent assay (ELISA) kit, the dilution of the patient's urine can be avoided or greatly reduced. The limit of detection was determined to be approximately 10 ng ml À1 , which is lower than the cutoff value for diagnosing bladder cancer (11.16 ng ml À1 ). When the values measured using the microfluidic chip were compared with those measured using conventional ELISA using a 96-well plate for five patients, the deviations were 0.9%, 6.8%, 9.4%, 1.8%, and 5.8%. The entire measurement time is 6-fold faster than that of conventional ELISA. This microfluidic device shows significant potential for point-ofcare applications. V C 2013 American Institute of Physics.

show abstract

“…The DNA was cloned into pCR TM 2.1-TOPO V R (Invitrogen, USA), total 33 colonies for HCT-8 specific peptide and 55 colonies for CRSC specific peptides were selected and checked by using plasmid extraction mini kit (FavorPrepTM Plasmid Extraction Kit/Favorgen, Taiwan) and PCR to ensure that each individual colony has DNA segment of interest. 16,24…”

Section: B Experimental Process Of On-chip Phage Displaymentioning

confidence: 99%

“…The K d value of the peptide was measured using flow cytometry (C6 Cytometer-BD Biosciences, BD Accuri TM , USA). 23,24 The cell number of the target cells was fixed to be 2 Â 10 5 and the concentration of the specific peptide used varied with 2-fold serial dilutions from 1 lM to 10 nM. Note that the binding site of the specific peptide was assumed to be only one.…”

Section: E Specificity Test and Dissociation Constant (K D ) Measurementioning

confidence: 99%

An integrated microfluidic system for screening of phage-displayed peptides specific to colon cancer cells and colon cancer stem cells

Jui

Hung

et al. 2015

Biomicrofluidics

Self Cite

View full text Add to dashboard Cite

Affinity reagents recognizing biomarkers specifically are essential components of clinical diagnostics and target therapeutics. However, conventional methods for screening of these reagents often have drawbacks such as large reagent consumption, the labor-intensive or time-consuming procedures, and the involvement of bulky or expensive equipment. Alternatively, microfluidic platforms could potentially automate the screening process within a shorter period of time and reduce reagent and sample consumption dramatically. It has been demonstrated recently that a subpopulation of tumor cells known as cancer stem cells possess high drug resistance and proliferation potential and are regarded as the main cause of metastasis. Therefore, a peptide that recognizes cancer stem cells and differentiates them from other cancer cells will be extremely useful in early diagnosis and target therapy. This study utilized M13 phage display technology to identify peptides that bind, respectively, to colon cancer cells and colon cancer stem cells using an integrated microfluidic system. In addition to positive selection, a negative selection process was integrated on the chip to achieve the selection of peptides of high affinity and specificity. We successfully screened three peptides specific to colon cancer cells and colon cancer stem cells, namely, HOLC-1, HOLC-2, and COLC-1, respectively, and their specificity was measured by the capture rate between target, control, and other cell lines. The capture rates are 43.40 ± 7.23%, 45.16 ± 7.12%, and 49.79 ± 5.34% for colon cancer cells and colon cancer stem cells, respectively, showing a higher specificity on target cells than on control and other cell lines. The developed technique may be promising for early diagnosis of cancer cells and target therapeutics.

show abstract

An automatic microfluidic system for rapid screening of cancer stem-like cell-specific aptamers

Cited by 40 publications

References 27 publications

Isolation of thermally sensitive protein-binding oligonucleotides on a microchip

Isolation of thermally sensitive protein-binding oligonucleotides on a microchip

A negative-pressure-driven microfluidic chip for the rapid detection of a bladder cancer biomarker in urine using bead-based enzyme-linked immunosorbent assay

An integrated microfluidic system for screening of phage-displayed peptides specific to colon cancer cells and colon cancer stem cells

Contact Info

Product

Resources

About