Periodic Reporting for period 1 - Biochip-images (A Breast Cancer Biomarker Point of CarePOCDiagnostic Platform Integrating Dual-Functionalized Nanoparticle for Magnetic Gradient Ranking and Electrochemical Sensing)
Periodo di rendicontazione: 2019-12-01 al 2021-11-30
This MSCA titled ‘Biochip-images’ for short aimed to implement novel approaches towards the development of a healthcare diagnostic assay capable of addressing the need for improved cross functional and adaptive therapeutics design. Now more than ever, it is clear that society stands to gain significantly from such advancements. Moreover, the recent and continued worldwide COVID-19 pandemic, and associated variants, have highlighted the clinical and societal benefits associated with early diagnosis and the importance of advanced assay development.
In this project, the MSCA Fellow focused on three major objectives: 1) surface functionalisation of magnetic NPs by coating with both antibodies and redox probes to develop dual-NPs for biomarker capture and electrochemical amplification, 2) understanding the mechanisms associated with integration of PDMS microfluidic channels, towards the constructions of a novel POC device for biomarker diagnostic associated with dual-NPs and 3) initial fabrication design concepts of Mag and Me arrays to develop a microchip for Mag ranking and electrochemical sensing of MP and MB.
This project has advanced the development of a new POC concept by integrating knowledge of magnetic nanoparticles (MP), advanced magnetophoresis, and a sensitive microelectrode sensing. Outcomes include;
1) Development of a novel method to isolate the target MP from the free MP, by use of a magnetic susceptibility gradient array.
2) The MP developed here has been labelled with both redox and biological properties, which allows for improved amplification as well as biomarker capture.
3) As a POC concept, the device could combine both the magnetic susceptibility gradient array and electrode array, enabling simplification of the sampling procedure and improved diagnostic efficiency (however, this would require further testing to verify).
4) Several Invention Discovery Applications (IDAs) have been submitted to the University.
A parallel objective of the MSCA Individual Fellowship is to foster the development and progression of the individual researcher which is also report on below.
In undertaking this MSCA, it has provided the Fellow with a framework to conduct this project within the top university of University College Dublin (UCD), working with access to world-class expertise within the Centre for Bio-Nano Interactions. Here the Fellow gained significant experience and knowledge not only within the core research objectives but also in project management, scientific writing, and proposal development.
Following the conclusion of MSCA, the Fellow has secured a role as a senior scientist in LumraDx based in the UK, and has been able to utilise the knowledge and skills gained during the project towards the development of COVID ultra diagnostic assays within the Research and Development Department.
In this project, the MSCA Fellow focused on three major objectives: 1) surface functionalisation of magnetic NPs by coating with both antibodies and redox probes to develop dual-NPs for biomarker capture and electrochemical amplification, 2) understanding the mechanisms associated with integration of PDMS microfluidic channels, towards the constructions of a novel POC device for biomarker diagnostic associated with dual-NPs and 3) initial fabrication design concepts of Mag and Me arrays to develop a microchip for Mag ranking and electrochemical sensing of MP and MB.
This project has advanced the development of a new POC concept by integrating knowledge of magnetic nanoparticles (MP), advanced magnetophoresis, and a sensitive microelectrode sensing. Outcomes include;
1) Development of a novel method to isolate the target MP from the free MP, by use of a magnetic susceptibility gradient array.
2) The MP developed here has been labelled with both redox and biological properties, which allows for improved amplification as well as biomarker capture.
3) As a POC concept, the device could combine both the magnetic susceptibility gradient array and electrode array, enabling simplification of the sampling procedure and improved diagnostic efficiency (however, this would require further testing to verify).
4) Several Invention Discovery Applications (IDAs) have been submitted to the University.
A parallel objective of the MSCA Individual Fellowship is to foster the development and progression of the individual researcher which is also report on below.
In undertaking this MSCA, it has provided the Fellow with a framework to conduct this project within the top university of University College Dublin (UCD), working with access to world-class expertise within the Centre for Bio-Nano Interactions. Here the Fellow gained significant experience and knowledge not only within the core research objectives but also in project management, scientific writing, and proposal development.
Following the conclusion of MSCA, the Fellow has secured a role as a senior scientist in LumraDx based in the UK, and has been able to utilise the knowledge and skills gained during the project towards the development of COVID ultra diagnostic assays within the Research and Development Department.
In WP2, the Fellow investigated the research area of magnetophoresis, by bibliography survey and aimed to advance the understanding of the magnetophoresis. Accordingly, a microscopy slide mini Luer platform template for the design was generated in Figure 1A-C.
WP3 considered a key element of this project which was the construction of dual-functionalized MPs via combination of an ultrasensitive redox probe and the specific task biological probes in order to realize the multi-functionalities of MPs in POC devices (such as magnetic manipulation, target capture and signal amplification. Three different strategies were investigated here;
MPs of γ-Fe2O3 dots were synthesized by co-precipitation and stabilized by surface functionalization process of amination and pegylation (Figure 2A). By cross-linking chemistry, the IgG as bio-probe model was conjugated onto the MP surface together with redox probe (Figure 2B) and electrochemical characterization (Figure 2C).
Secondly, a facile synthetic strategy was also used to fabricate such multi-functionalized MPs. TEM images (Figure 3A) of these MPs after coating of three polyelectrolyte layers were characterized to be 11nm of MPs@Fc, while the redox intensity of the three MPs solution at 1.0 mg.mL-1 were electrochemically detected to be around 0.64µA(Figure 3B). Moreover, the binding efficiency of IgG was investigated here, where the IgG was attached to the PSS terminated redox MPs to form biological corona nanoparticle (Figure 3C)
Thirdly, development and learning of surface engineering techniques facilitated the possibility to combine redox and biological properties onto MPs surface. Here, the Fellow developed multi-core MPs (Figure 4A) with a novel strategy to advance redox MPs by covalently conjugated BSA and redox probes (Figure 4B). IgG was efficiently binding onto MPs surface by click chemistry (Figure 4C).
WP4 incorporated the central concept to isolate targets based MPs and free MP, the Fellow initiated work on the design of an advanced microfluidic chip based on magnetic susceptibility gradient (MsG) ranking technique, connected to WP3 & WP2. To complete the novel POC concept, a micro-electrode sensing array was also designed inside the MsG area so that each MP or MP based targets flowing through the sensing electrode can be electrochemically detected.
In WP5, the successfully developed nanoparticles featured with biological, redox and magnetic properties were used for bio-application in order to assess the MPs in a clinical application, Here, exosomes collected from A549 cells culture were used to evaluate the MPs’ function, such as selective bio-capture, efficient magnetic separation, and sensitively electrochemical sensing have collected exosome. In addition, CD63 antigen was also used as a biomarker, to confirm their bio-affinity.
On completion of the project, the Fellow presented to the research group within CBNI an overview and outcomes, key findings and knowledge gained as part of the MSCA Action (Figure 5) and ensured transfer of knowledge with colleagues and students.
WP3 considered a key element of this project which was the construction of dual-functionalized MPs via combination of an ultrasensitive redox probe and the specific task biological probes in order to realize the multi-functionalities of MPs in POC devices (such as magnetic manipulation, target capture and signal amplification. Three different strategies were investigated here;
MPs of γ-Fe2O3 dots were synthesized by co-precipitation and stabilized by surface functionalization process of amination and pegylation (Figure 2A). By cross-linking chemistry, the IgG as bio-probe model was conjugated onto the MP surface together with redox probe (Figure 2B) and electrochemical characterization (Figure 2C).
Secondly, a facile synthetic strategy was also used to fabricate such multi-functionalized MPs. TEM images (Figure 3A) of these MPs after coating of three polyelectrolyte layers were characterized to be 11nm of MPs@Fc, while the redox intensity of the three MPs solution at 1.0 mg.mL-1 were electrochemically detected to be around 0.64µA(Figure 3B). Moreover, the binding efficiency of IgG was investigated here, where the IgG was attached to the PSS terminated redox MPs to form biological corona nanoparticle (Figure 3C)
Thirdly, development and learning of surface engineering techniques facilitated the possibility to combine redox and biological properties onto MPs surface. Here, the Fellow developed multi-core MPs (Figure 4A) with a novel strategy to advance redox MPs by covalently conjugated BSA and redox probes (Figure 4B). IgG was efficiently binding onto MPs surface by click chemistry (Figure 4C).
WP4 incorporated the central concept to isolate targets based MPs and free MP, the Fellow initiated work on the design of an advanced microfluidic chip based on magnetic susceptibility gradient (MsG) ranking technique, connected to WP3 & WP2. To complete the novel POC concept, a micro-electrode sensing array was also designed inside the MsG area so that each MP or MP based targets flowing through the sensing electrode can be electrochemically detected.
In WP5, the successfully developed nanoparticles featured with biological, redox and magnetic properties were used for bio-application in order to assess the MPs in a clinical application, Here, exosomes collected from A549 cells culture were used to evaluate the MPs’ function, such as selective bio-capture, efficient magnetic separation, and sensitively electrochemical sensing have collected exosome. In addition, CD63 antigen was also used as a biomarker, to confirm their bio-affinity.
On completion of the project, the Fellow presented to the research group within CBNI an overview and outcomes, key findings and knowledge gained as part of the MSCA Action (Figure 5) and ensured transfer of knowledge with colleagues and students.
The MSCA has aimed to advance our understanding in state-of-the-art magnetophoresis, electroanalysis, nanotechnology, and microfluidic integration towards the development of a new point of care (POC) device for biomarker detection.
Socio-economic impact
The potential societal impact of this project is clear. Advancements in diagnostic technology will be key to delivering next generation therapeutics for future generations. As highlighted above, cancers continue to impact significantly on European communities with 25% of all annual cancer cases being attributed to the region. Moreover, it is estimated that this rate is likely to increase as well as the associated economic costs estimated at €199 Billion in 2018.
Wider society implications
This project has aimed at development of key knowledge and technologies towards the design of a POC assay. From the POC concept design, it is envisaged that the POC device will be versatile. As the specific bio-probe was combined onto MP surface, by change the bio-probes, the POC device can not only be used for cancer biomarker diagnosis but may also be used in a cross functional diagnostic application for other illnesses such as COVID-19. Compared to the traditional POC diagnostic device, this POC concept integrates nanoparticle, magnetophoresis and electrochemical sensing while also exhibiting other advantages including simple sampling process, highly versatile assay, and high detection sensitivity.
Socio-economic impact
The potential societal impact of this project is clear. Advancements in diagnostic technology will be key to delivering next generation therapeutics for future generations. As highlighted above, cancers continue to impact significantly on European communities with 25% of all annual cancer cases being attributed to the region. Moreover, it is estimated that this rate is likely to increase as well as the associated economic costs estimated at €199 Billion in 2018.
Wider society implications
This project has aimed at development of key knowledge and technologies towards the design of a POC assay. From the POC concept design, it is envisaged that the POC device will be versatile. As the specific bio-probe was combined onto MP surface, by change the bio-probes, the POC device can not only be used for cancer biomarker diagnosis but may also be used in a cross functional diagnostic application for other illnesses such as COVID-19. Compared to the traditional POC diagnostic device, this POC concept integrates nanoparticle, magnetophoresis and electrochemical sensing while also exhibiting other advantages including simple sampling process, highly versatile assay, and high detection sensitivity.