Posts

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

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

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


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.

Bio Convention San Francisco

Meet us at Bio Convention in San Francisco, US

AWSensors will join the Bio Convention next week in San Francisco, US. You may meet us at Spanish Pavilion in booth 605 or in the one-on-one partnering in the Bio Business Forum.

Prof. Antonio Arnau will attend the convention to explain how AWS sensing technology works and its applications in health or agri-food research. Furthermore he will introduce the european project LiqBiopSens and how our sensing technology will be used to developed a liquid biopsy platform for early detection of colorectal cancer.

The Bio International Convention (BIO) attracts over 15,000 biotechnology leaders. We will spend one week of intensive networking to discover new opportunities and promising partnerships with some biotech, pharma and industry leaders and several academic institutions that are attending the most important event in the biotech industry worldwide.. This event covers a wide spectrum of life science and application areas including nanotechnology, drug discovery, genomics, cell therapy or biofuels.

70% out of R+D in bio field is conducted in US where there are more than 2500 companies in this sector.

We are proud to attend the event along with other Spanish biotech companies, a group of about 70 companies together in the Spanish pavilion as you can look up at the SpainBio catalogue.

If you would like to meet with Prof. Arnau in San Francisco from june 6 to 9, do not hesitate to contact us!

AWSensors at Biosensors 2016

AWSensors technology at Biosensors 2016

Visit the booth of our distributor, Bio-Logic, to see our new products

Just one week left for Biosensors 2016! AWSensors technology will be at the most prestigious congress on biosensing research to be held at Gothenburg, Sweden from May 25th to 26th. We are pleased to invite you to see our platforms at the booth of Bio-Logic, our OEM distributor. There, you will see how our platform works.

Our equipment is the only quartz crystal microbalance that allows:

–           Detection and characterization of interface phenomena with high sensitivity (up to 0.05 ng/cm2)

–           Use of different kind of sensors simultaneously.

Our technology is based on two fundamental principles:

  1. Use of high fundamental frequency acoustic resonators (100-150 MHz) that provides a large increase in sensitivity.
  2. Use of a novel method for characterizing these sensors which keeps noise at very low level.

As a result, an improvement in the limit of detection (LOD) of 2 orders of magnitude compared to traditional acoustic sensors is obtained.

Do not miss the opportunity to check the advantages of our new technology and meet AWSensors founder, Prof. Antonio Arnau. He is coordinator of the International Network of Piezoelectric Transducers Research and Applications (PETRA) and has wide expertise on bioelectronics and acoustic wave biosensors.

Dr. Arnau will attend the Post Congress Symposium in Cancer Diagnostics to be held on Saturday 28 May inmediately following Biosensors 2016. AWSensors is currently coordinating LiqBiopSens European Project to develop a new liquid biopsy platform for early detection of colorectal cancer.

Bioeconomy in Spain

Bioeconomy in the Spanish Public Budget for 2016

Spanish Parliament starts to debate today the public budget for the next year. So that, we would like to review what are the plans to invest in research and innovation. According to the budget for the Secretary of Innovation, bioeconomy will be one of the priorities for the Spanish Government in 2016.

 

Bioeconomy figure; food, biomass, public health, energy security, sustainable production, economic and social development

What is bioeconomy?

The bioeconomy definition according to the EFIB (European Forum for the Bioeconomy) is the following:

Bioeconomy encompasses the production of renewable biological resources and their conversion into food, feed, bio-based products and bioenergy via innovative and efficient technologies.

AWSensors tech has important applications in bioeconomy such as environmental monitoring, secure food and water, bioenergy or biomaterials. EU is promoting and funding projects in these fields in order to make the transition to a more resource efficient society and the Spanish Government is following this policy too.

Spain will boost the Bioeconomy Strategy in the public budget to research and development for 2016

Spanish government plans to invest 1,765 million euros in direct aid for innovation projects and 2,446 million euros for loans to enterprises. The resources allocated to direct aid will increase up to 15 per cent over this year and loans will decrease to 9 per cent. The government will give priority to areas considered most in need of development in science and technology such as Andalucia, Extremadura, Canarias, Castilla-La Mancha, Murcia, Galicia and Asturias. Among the actions to be undertaken in these regions, Bioeconomy will be boosted in the agri-food, biomass, bioenergy and bioproducts fields.

Furthermore, the Centre for the Development of Industrial Technology (CDTI), the main driver of innovation in Spain, will have 30 million euros more for direct aid, 35.7% more than the previous year. The Spanish High Council Scientific Research (CSIC) will have 467 million euros next year and the Institute of Health Carlos III (ISCIII), 165 millions.

In spite of this state budget for 2016, Spain is still the 17th european country in gross domestic expenditure on R&D according to Eurostat.

Further sensitivity enhancement of HFF-QCM immunosensors for pesticides

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