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Liqbiopsens project

AWSENSORS HOSTS THE 24M MEETING OF LIQBIOPSENS EUROPEAN PROJECT

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February 1st 2018

AWSensors host the 24-Month Meeting of LIQBIOPSENS EU project

The meeting of all the project partners was held in Valencia, Spain, at the headquarters of AWSensors and Sistemas Genómicos, where significant advances were presented between the project partners and important decisions were taken to successfully continue pursuing the project goals. Another whole year is still required to complete the project.

The consortium for the development of this project is composed of the following institutions: Servicio Andaluz de Salud (Spain), Université Catholique de Louvin (Belgium),  Foundation for Research and Technology Hellas (Greec), DestiNA Genomics (UK, Spain), Sistemas Genómicos (Spain) and AWSensors (Spain). AWSesensor.

If you wish to know more about the LIQBIOPSENS project, visit the project website at http://liqbiopsens.com.

 

Design and Validation of a 150 MHz HFFQCM Sensor for Bio-Sensing Applications

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Authors: Román Fernández1, Pablo García1, María García1, José V. García1, Yolanda Jiménez2 and Antonio Arnau2

1Advanced Wave Sensors S. L., Algepser 24, 46988 Paterna, Spain
2Centro de Investigación e Innovación en Bioingeniería, Universitat Politècnica de València, Camí de Vera S/N, 46022 Valencia, Spain

Journal: Sensors Journal (2017)

Abstract: Acoustic wave resonators have become suitable devices for a broad range of sensing applications due to their sensitivity, low cost, and integration capability, which are all factors that meet the requirements for the resonators to be used as sensing elements for portable point of care (PoC) platforms. In this work, the design, characterization, and validation of a 150 MHz high fundamental frequency quartz crystal microbalance (HFF-QCM) sensor for bio-sensing applications are introduced. Finite element method (FEM) simulations of the proposed design are in good agreement with the electrical characterization of the manufactured resonators. The sensor is also validated for bio-sensing applications. For this purpose, a specific sensor cell was designed and manufactured that addresses the critical requirements associated with this type of sensor and application. Due to the small sensing area and the sensor’s fragility, these requirements include a low-volume flow chamber in the nanoliter range, and a system approach that provides the appropriate pressure control for assuring liquid confinement while maintaining the integrity of the sensor with a good base line stability and easy sensor replacement. The sensor characteristics make it suitable for consideration as the elemental part of a sensor matrix in a multichannel platform for point of care applications.

Keywords: HFF-QCM (high fundamental frequency quartz crystal microbalance); finite element method (FEM); flow cell; biosensor; PoC (point of care); MQCM (monolithic quartz crystal microbalance)

 Sensors 17 02057 g002 550

You can download the full paper here

Biotechnology business

AWSENSORS RAISED 1M€ INVESTMENT

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July 27th 2017

Swedish and Spanish companies invested in AWSensors seeing a growth opportunity.

AWSensors raised 1M€ investment round to strengthen its position in scientific market and enter the healthcare tech market, one of the most competitive markets. The company though has convinced Swedish and Spanish investors that the potential payoff is worth the risk.

Firms entering into the investment round were BAble Capital, a venture capital firm that aims to invest in technology-based companies from Spanish universities and research centers, and Tech Transfer UPV, a venture capital firm created specifically to transfer technology developed at the Spanish university, Polytechnic University of Valencia. But also a consortium of Swedish companies invested in AWSensors seeing a growth opportunity. Other Spanish companies like PolymerChar, Keodes or Citrosol are also among the shareholders.

Prof. Arnau, founder of AWSensors

AWSensors, led by Prof. Antonio Arnau, expects to speed up innovation processes to launch new scientific equipments to meet demand from scientists and industry R&D but also to develop cutting-edge healthcare technology to be applied in personalized medicine.

The aim of these new developments is to get a blood test meant to catch cancer when it’s most treatable, before patients show symptoms, by detecting fragments of DNA shed by tumors.  A new liquid biopsy system will allow for detection of this circulating DNA and its mutations and will be an essential equipment to help doctors diagnose, monitor and treat cancer patients in a personalized way.

To hit that goal, AWSensors got another 1M€ from European Comission through two H2020 European projects:

  • LiqBiopSens project, that is coordinated by AWSensors and developed together with companies and institutions from Spain, Belgium, UK and Greece.
  • Catch-u-DNA project, which is carried out with partners from Germany, France, Israel, Greece and Spain.

These projects as a whole were funded with 5.7M€ by European Comission.

AWSENSORS CONSIGUE UNA INVERSIÓN DE 1 MILLÓN DE EUROS

Nuevos inversores de Suecia y España entran en el accionariado de la compañía

AWSensors ha conseguido despertar el interés de inversores nacionales e internacionales y realizar una ronda de 1 millón de euros. En la operación de inversión han participado BeAble Capital, sociedad gestora que tiene como objetivo invertir en empresas de base tecnológica con origen en universidades y centros de investigación españoles, y Tech Transfer UPV, fondo creado específicamente para trasladar al mercado tecnología desarrollada en la Universidad Politécnica de Valencia. Pero también un consorcio de empresas tecnológicas suecas ha apostado fuerte por entrar como inversor en AWSensors. Además, empresas valencianas como PolymerChar, Keodes o Citrosol están también entre los accionistas.

Esta inversión permitirá a AWSensors consolidarse en el mercado científico internacional donde comercializa una nueva tecnología patentada de sensores para análisis de interacciones moleculares en tiempo real que utilizan investigadores de Europa, Estados Unidos y Asia en los campos de ciencias de la vida y nuevos materiales.

La compañía, liderada por el Prof. Antonio Arnau, espera invertir en nueva infraestructura de fabricación y captar nuevo talento para su plantilla. El objetivo es acelerar el proceso de innovación para sacar al mercado nuevos equipos con mayor productividad y automatización e iniciar nuevos desarrollos para el mercado de salud, ya que la tecnología de AWSensors podrá ser aplicada en medicina personalizada.

Uno de los desarrollos previstos para el mercado sanitario es un equipo para la detección precoz de cáncer colorrectal y su monitorización sin necesidad de entrar al quirófano para realizar una biopsia del tumor. Servirá con un análisis de sangre cuyos resultados se podrían obtener en una hora. La detección en este caso se basa en el ADN que libera el tumor en el organismo. El nuevo sistema permitirá detectar ese ADN circulante y las mutaciones que sufre asociadas al cáncer. Este análisis, denominado biopsia líquida, permitirá un control sencillo del paciente y facilitará por tanto la adaptación del tratamiento de forma personalizada.

Liqbiopsens project

Este nuevo equipo se está desarrollando junto a otras empresas e instituciones de España, Bélgica, Reino Unido y Grecia dentro del proyecto LiqBiopSens financiado por la Comisión Europea dentro del programa H2020. Un proyecto que ha inyectado medio millón de euros a AWSensors.

Paralelamente a este desarrollo, AWSensors participa en otro proyecto europeo, Catch-u-DNA que investiga una nueva técnica para mejorar la sensibilidad de la tecnología en la detección del ADN circulante. Este proyecto, que se realiza con socios de Alemania, Francia, Israel, Grecia y España, ha supuesto el ingreso de medio millón de euros más en la compañía.

En total, la Comisión Europea ha dotado con 5’7 millones de euros a estos dos proyectos de investigación cuyos resultados impulsarán el crecimiento de AWSensor

Catch-u-DNA project towards a new technology for molecular diagnosis

New project to validate an easier and faster technique of molecular diagnosis applied to cancer by using quartz sensors

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July 7th 2017

In 2015 AWSensors already got the European project “LiqBiopSens” to develop a liquid biopsy platform for early detection and monitoring of colorectal cancer. Now, the company will work in the European  project “CATCH-U-DNA”, worth 3.4 million euros. CATCH-U-DNA poses a new concept in biophysics and molecular diagnosis based on acoustic sensing of DNA in serum. It aims to provide a novel technology for the ultrasensitive detection of circulating-tumor DNA in serum without PCR amplification.

“CATCH-U-DNA” project was granted under the Horizon 2020 FET-OPEN call, aimed at financing radically new initiatives that can have a long-term economic and social impact. Only the most cutting-edge technology projects are chosen. It is one of the most competitive European programs, where less than 4 per cent of the ideas presented are funded. The project is coordinated by Prof. Electra Gizeli, leader of the Biosensors group in the Institute of Molecular Biology and Biotechnology of the Greek Foundation for Research and Technology-Hellas (FORTH).

Detection of lung and colorectal cancer

The ultimate goal of “CATCH-U-DNA” is to validate a new, simpler and cheaper technology for the detection of genetic markers so that personalized medical diagnosis can be performed in a more precise, easy and affordable way than current technologies.

The project faces the ambitious challenge of testing that radically new technology in the early diagnosis of cancer by detecting the most common genetic mutations that cause colorectal cancer and lung cancer. In fact, it includes not only experimental research but also clinical trials.

In the future, it will be possible to incorporate this new technique into portable equipment and will therefore allow on-site analysis applied to personalized medicine in developed countries or in areas without laboratory infrastructure such as underdeveloped countries.

New technique easier, more affordable and more precise

The amount of mutated DNA in a sample is generally very low, which greatly hampers its detection and characterization. Most of the current methods of detection are based on the technique known as PCR (polymerase chain reaction), which allows amplifying a fragment of DNA by obtaining millions of copies, thus, its detection in the lab is feasible. However, this technique involves a complex and expensive procedure prior to detection. In addition, it may cause a deviation if the amplification is not developed properly.

The “CATCH-U-DNA” technology promises to overcome these limitations and allow for the genetic analysis of human samples in an easier, more affordable and more precise way. The aim is to manufacture an ultra-sensitive device capable of detecting DNA in human samples without any previous amplification procedure even though the amount of that DNA is minimal. Specifically, the goal is the detection of circulating DNA, fragments from the tumor cells found in the blood.

The new technique will use high frequency quartz crystal sensor arrays that allow for a real-time, label-free monitoring, in combination with a revolutionary DNA identification system based on the hydrodynamic molecular properties. This approach will allow to push the detection limit down obviating the amplification.

7 companies and organizations from 5 different countries

The “CATCH-U-DNA” project will be developed over 3 years. Experts in molecular biology, physics, chemistry, nanomaterials, biosensors and microfluidics from Spain, Germany, France, Greece and Israel are involved. They are from the Greek Foundation for Research and Technology Hellas (FORTH), which is the project coordinator, and from AWSensors; the Autonomous University of Madrid, in Spain; the University of Crete in Greece; the Curie Institute in France; the Ben Gurion University of Negev in Israel and the German company Jobst Technologies.

 

A liquid biopsy platform combining a high fundamental frequency QCM device with dynamic chemistry for detecting mutations in circulating DNA

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Authors: A. Grammoustianou, G. Papadakis, M. Tabraue, J.J. Díaz-Mochon, R. Fernández, J.V. García, A. Arnau, E. Gizeli. AWSensors S.L., Institute of Molecular Biology and Biotechnology- FORTH, University of Crete, Destina Genomics S.L., Centro de Investigación e Innovación en Bioingeniería – Universidad Politécnica de Valencia

Event:  5th International Conference on Bio-sensing Technology, Riva del Garda, Italy (2017)

In the past decade, the analysis of circulating tumour DNA (ctDNA) in blood has been a major breakthrough; ctDNA has been proposed as a priceless source for cancer diagnostic, prognostic and treatment monitoring through a new methodology known as “Liquid Biopsy”. This study presents a novel diagnostic method for the acoustic detection of KRAS mutations in ctDNAs based on: (1) DNA analysis by “dynamic chemistry” that utilizes aldehyde modified nucleobases (SMART) and abasic peptide nucleic acids (DGL probes) capable for the errorfree detection of nucleic acids and their mutations; and, (2) a high fundamental frequency (100 MHz) acoustic wave microsensor (AWS HFF-QCM) that allows the accurate, inexpensive, label-free and real time monitoring of the “dynamic chemistry”. Surface-immobilized DGL probes on the AWS HFF-QCM device are used to detect ctDNAs of wild type and mutated KRAS variants. Upon hybridization of the DGL probe with its target ssDNA, a duplex is formed where biotin- tagged SMART bases can lock in front of the position under interrogation; streptavidin binding detected in a follow-up step confirms the presence of the SMART bases. The use of DGL probes in combination with an isothermal DNA amplification step RPA) have allowed the sensitive and specific recognition of single mismatches in KRAS genes in less than 1 hour. This work presents a unique and novel technology that can emerge as a promising tool in the field of cancer diagnostics.

Liquid Biopsy detection protocol

Schema for complete detection protocol:

(A) Extracted DNA containing mutant (red) and wild type DNA fragments (black) are enzymatically amplified.

(B) Denatured amplicons are hybridized on surface immobilized DGL probes. Chemical locking of a specific tagged SMART base takes place only in the appropriate position.

(C) Incorporated SMART bases are recognized by streptavidin and monitored in real-time during an acoustic measurement.

A High Fundamental Frequency QCM Immunosensor for Tuberculosis Detection

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AWSensors biosensor for tuberculosis detection

Authors: Ángel Montoya, Carmen March, Yeison J. Montagut, Maria J. Moreno, Juan J. Manclus, Antonio Arnau, Yolanda Jiménez, Marisol Jaramillo, Paula A. Marin, Robinson A. Torres

Journal:  Current topics in Medicinal Chemistry (2017)

Background: Tuberculosis, one of the oldest diseases affecting human beings, is still considered as a world public health problem by the World Health Organization.

Method & Material: Therefore, there is a need for new and more powerful analytical methods for early illness diagnosis. With this idea in mind, the development of a High Fundamental Frequency (HFF) piezoelectric immunosensor for the sensitive detection of tuberculosis was undertaken. A 38 kDa protein secreted by Mycobacterium tuberculosis was first selected as the target biomarker. Then, specific monoclonal antibodies (MAbs) were obtained. Myc-31 MAb, which showed the highest affinity to the analyte, was employed to set up a reference enzyme-linked immunosorbent assay (ELISA) with a limit of detection of 14 ng mL-1 of 38 kDa antigen.

Results & Discussion: For the development of the HFF piezoelectric immunosensor, 100 MHz AWSensors quartz crystals were used as transducer elements. The gold electrode surface was functionalized by covalent immobilization of the target biomarker through mixed self-assembled monolayers (mSAM) of carboxylic alkane thiols. A competitive immunoassay based on Myc-31 MAb was integrated with the transducer as sensing bio-recognition event. Reliable assay signals were obtained using low concentrations of antigen for functionalization and MAb for the competitive immunoassay. Under optimized conditions, the HFF immunosensor calibration curve for 38 kDa determination showed a limit of detection as low as 11 ng mL-1 of the biomarker. The high detectability attained by this immunosensor, in the picomolar range, makes it a promising tool for the easy, direct and sensitive detection of the tuberculosis biomarker in biological fluids such as sputum.

You may review the full paper here

Bio-Logic AWS-A20 at Biosensing Conference

AWSensors at the 5th International Conference on Biosensing technology

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May 4th 2017

AWSensors technology will be at the  5th International Conference on Bio-sensing technology to be held at Riva del Garda, Italy, from May 7th to 10th. We are pleased to invite you to see our biosensing platforms at the booth of Bio-Logic, our OEM distributor.

AWS A20-F20 platform is the most flexible and sensitive QCM-D system in the market. It is able to work with low and high frequency QCM sensors and SAW sensors. It also ensures high sensitivity because of a novel method of characterization that keeps noise at very low level in spite of working at high frequencies by using AWS-HFF sensors (100-150 MHz). AWSensors also provides QCM and SAW sensors and a wide range of cells and accessories to customize experiments and tests according to specific conditions required by scientists.

Therefore, AWS A20 system provides you with big competitive advantages:

–       High flexibility, capable to work with any acoustic wave sensor; modular system capable to be upgraded with more channels after purchase (up to 4)

–       High sensitivity

–       Customization, with accessories to make measurements in specific conditions

–       Price convenience

Do not miss the opportunity to check the advantages of our technology at the booth of Bio-Logic and meet our Application Scientist, Mrs. María García.

Dynamic Resolution of Ion Transfer in Electrochemically Reduced Graphene Oxides Revealed by Electrogravimetric Impedance

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Authors: Hamza Goubaa, Freddy Escobar-Teran, Ibtissam Ressam, Wanli Gao, Abdelkrim el Kadib, Ivan T. Lucas, Mustapha Raihane, Mohammed Lahcini, Hubert Perrot, Ozlem Sel. Sorbonne Université, UPMC, Univ Paris 06, CNRS, France. Laboratoire Interfaces et Systèmes Électroquimiques; Université Cadi Ayyad, Faculté des Sciences et Techniques, Laboratoire de Chimie Organométallique et Macromoléculaire Matériaux composites, Marrakech, Morocco.

Journal:  The Journal of Physical Chemistry (2017)

To accompany the search for optimal materials in electrochemical supercapacitors, appropriate characterization tools to assess key parameters of newly developed electrodes are required. Here we demonstrate the capabilities of ac-mode electrogravimetry to study in details the capacitive charge storage mechanisms in electrochemically reduced graphene oxide (ERGO) thin films electrodes. The coupling of electrochemical impedance spectroscopy (EIS) with fast quartz crystal microbalance (QCM) complements classical electrochemical quartz crystal microbalance (EQCM) by capturing here the dynamics of the electroadsorption process, identifying charged moieties and detecting solvation effects. We evidenced the co-electroadsorption of two types of cationic species (fully and partially hydrated cations) in the potential range studied and the indirect intervention of free solvent molecules. Further kinetic information on electroadsorption is also obtained which leads to a full deconvolution of the global EQCM response at both gravimetric and temporal level. Through a fine analysis of the interactions of different cations of period one elements, Li+, Na+ and K+ with ERGO, we evidenced the critical role of solvation processes on the kinetics of electroadsorption, and provided an experimental proof to the phenomena that smaller ions are more tightly bound to their water molecules such that an inverse relationship exists between the dehydration energy and the ion size. Such gravimetric and dynamic subleties are unreachable with classical tools and with EQCM method alone which permits us to suggest the ac-mode electrogravimetry as a baseline diagnostic tool to explore charge transfer mechanisms at the nanoscale.

Further sensitivity enhancement of HFF-QCM immunosensors for pesticides

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Authors: C. March, J.V. García, R. Fernández, Y.Jiménez, A. Arnau. A.Montoya

Event: 4th International Conference on Biosensing Technology, Lisbon (2015)

Recently, High Fundamental Frequency Quartz Crystal Microbalance (HFF-QCM) immunosensors have succesfully been developed. Therefore, sensitivity of QCM biosensors is no longer a drawback. Taking advantage of this previous work, we have developed a renewed highly sensitive HFF piezoelectric immunosensor using carbaryl insecticide as a model analyte for pesticide detection. To this purpose, 100 MHz quartz crystal sensors were used as the transducer elements of the biosensor and a monoclonal antibody-based competitive immunoassay was integrated as the sensing specific bio-recognition event. The biosensing interface was improved by employing mixed self-assembled monolayers (mSAMs) of alkane thiols as intermediate layers for surface functionalization. This approach allowed the covalent attachment of the assay conjugate (20.0 µg mL-¹ of BSA-CNH conjugate) onto the gold electrode surface in a more orderly and stable way than with simple SAMs. A very low concentration (1.0 µg mL-¹) of LIB-CNH45 monoclonal antibody was used for the competitive immunoassays. All immunosensor assays were performed in the AWS-A10 test platform from AWSensors. In terms of analytical performance, the new carbaryl HFF-QCM immunosensor showed higher sensitivity than the previously developed one, with analytical parameters very close to those of the most sensitive reported ELISA for carbaryl.

CLICK HERE TO DOWNLOAD THE POSTER about this work

Comparison between High Fundamental Frequency Quartz Crystal Microbalance and Love Mode Surface Acoustic Wave devices in the detection of Carbaryl pesticide

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Authors: J.V. García, M.I. Rocha, C. March, P. García, L.A. Francis, A. Montoya, A. Arnau, Y. Jiménez

Event: Acoustic Sensors in analytical and Biophysical Studies, 29-30 August 2013, IMBB-FORTH, Heraklion, Crete.

 

In this work two acoustic technologies: High Fundamental Frequency Quartz Crystal Microbalance (HFF-QCM) and Love Mode Surface Acoustic Wave (LM-SAW) have been compared for the detection of Low Molecular Weight (LMW) compounds in terms of Sensitivity and LOD. The results have also been compared with those obtained with other techniques: Traditional QCM, Surface Plasmon Resonance (SPR) and Enzyme-Linked ImmunoSorbent Assay (ELISA). Carbaryl pesticide was chosen as model analyte because it had been used as a reference LMW compound by those mentioned techniques.

AWS-A10 research platform (AWSensors, Spain) was used to perform the experiments. This platform allowed for a comparison of both devices measured by the same characterization system under similar experimental conditions.

The results achieved with LM-SAW and 100Mhz HFF-QCM were in the same order of magnitude. The achieved value sensitivity (I50 value) and LOD (I90 ) were around 0.31 µ/L and 0.09 µ/L, respectively, for LM-SAW device, and around 0’66 µ/L and 0’14 µ/L, respectively, for HFF-QCM device. Sensitivities and LODs with these novel developments on acoustic technologies improve SPR and traditional QCM technologies, and approach ELISA’s ones.

Review the poster about this experiment by clicking here:

Póster about AWS-HFF sensors for detection of Carbaryl pesticide showed at Crete 2013