Periodic Reporting for period 1 - EV FOx (Extracellular Vesicles Fiber Optic Surface Plasmon Resonance selection and analysis)
Reporting period: 2023-07-01 to 2024-06-30
The Extracellular vesicles (EV) field is growing due to increased interest in their roles in diagnostics and therapeutics. Current methods to purify and quantify EV’s take a long time, introduce a bias, are costly and imprecise, thereby hampering advancement in the EV R&D field, while the lack of methods to quantify EVs directly in crude samples restricts applications in diagnostic and bioprocessing.
The main pain for EV R&D community (both academic and industrial) is a lack of standardized EV tools that allow a user-friendly, time-and cost-efficient EV isolation, quantification and characterization. Inconsistency in non-standardized methodologies for the reproducible isolation, quantification and characterization of EVs is a major
pain that hampers rapid advancement in EV research. Diagnostics save lives in cancer and cardiovascular diseases. Earlier diagnosis reduces testing need and disease severity, while stratification efficiently selects an effective treatment. Extracellular vesicles (EV) show potential in significantly reducing the associated healthcare costs by >5% but method inconsistency for EV isolation and analysis in crude samples is hampering progress in EV R&D and diagnostics.
Current solutions are characterized by high variability, poor sensitivity, biased sample preparation and characterization, long, unreproducible, and inaccurate protocols blocking accurate and unbiased EV isolation and quantification analysis. EV isolation is traditionally based on filters and/or ultracentrifugation (UC). These protocols are labor-intensive, variable and slow with questionable integrity and purity. These existing solutions were either not specifically designed for EV handling and characterization, or fall short in being able to handle the sample matrices, the performance of the solution and the ability to link the different characterization techniques. This unreliable EV quantification obtained from biased samples restricts EV use in R&D and diagnostics to qualitative decisions at best. Current EV quantification and characterization tools thus impair clinical insights. There is a pressing need to quantify EV during isolation and quantify their molecular contents to provide relevant information. KOLs in the EV field, involved in this project, are aware of this and at the forefront of addressing this.
The main pain for EV R&D community (both academic and industrial) is a lack of standardized EV tools that allow a user-friendly, time-and cost-efficient EV isolation, quantification and characterization. Inconsistency in non-standardized methodologies for the reproducible isolation, quantification and characterization of EVs is a major
pain that hampers rapid advancement in EV research. Diagnostics save lives in cancer and cardiovascular diseases. Earlier diagnosis reduces testing need and disease severity, while stratification efficiently selects an effective treatment. Extracellular vesicles (EV) show potential in significantly reducing the associated healthcare costs by >5% but method inconsistency for EV isolation and analysis in crude samples is hampering progress in EV R&D and diagnostics.
Current solutions are characterized by high variability, poor sensitivity, biased sample preparation and characterization, long, unreproducible, and inaccurate protocols blocking accurate and unbiased EV isolation and quantification analysis. EV isolation is traditionally based on filters and/or ultracentrifugation (UC). These protocols are labor-intensive, variable and slow with questionable integrity and purity. These existing solutions were either not specifically designed for EV handling and characterization, or fall short in being able to handle the sample matrices, the performance of the solution and the ability to link the different characterization techniques. This unreliable EV quantification obtained from biased samples restricts EV use in R&D and diagnostics to qualitative decisions at best. Current EV quantification and characterization tools thus impair clinical insights. There is a pressing need to quantify EV during isolation and quantify their molecular contents to provide relevant information. KOLs in the EV field, involved in this project, are aware of this and at the forefront of addressing this.
The establishment of a kit for isolating and releasing total EVs with high purity (>95%) and low hands-on time (< 20 min). Activities focus on the late stage development of:
• the sensor probe which captures the total EV population with high affinity. A streptavidin probe will be developed for capturing EV subpopulations.
• Reagents and protocols for isolating and releasing EVs with high integrity for mass spectrometry (high EV
concentration) and electron microscopy (low EV concentration) characterisations.
The main achievements are capture antibody coated probes, an efficient EV isolation and release method
and kit, and finally a validated EV isolation and release methodology and kit using real samples.
The establishment of a total EV quantitation assay for the White Fox instrument able to quantitate the total EV
concentration in crude samples, i.e. in first instance applied to plasma and cell culture media.
The main achievements here are the EV quantitation assay and assay reagents, a specific EV quantitation analysis algorithm, and validation of EV quantification method using real samples from KOL.
Extend the current White FOx software capabilities to include features for supporting EV analysis applications, such as EV sample information, instrument controls for EV sample handling, real-time feedback on EV isolation and release, and EV quantitation. Tweak the software to support GLP and GMP users by upgrading the software to meet 21 CFR part 11 requirements. The main achievements are installers and all supporting documentation for these 2 software solutions.
The objective is to realize an effective go-to-market strategy to drive commercialisation of FOx EV applications, consumables and products. FOx will therefore develop a Marketing and Sales plan, based on further market assessment, including a Product Launch Readiness plan. FOx as an organisation also needs to scale-up operationally to support the commercialisation of FOx EV applications and products, for which an internal operational scale-up and HR strategy will be developed. The main achievements are a Design and Development Plan, including risk assessment/ mitigation plan & User Requirement specifications, an IP management and dissemination plan, and all other operational supporting and planning documentation.
• the sensor probe which captures the total EV population with high affinity. A streptavidin probe will be developed for capturing EV subpopulations.
• Reagents and protocols for isolating and releasing EVs with high integrity for mass spectrometry (high EV
concentration) and electron microscopy (low EV concentration) characterisations.
The main achievements are capture antibody coated probes, an efficient EV isolation and release method
and kit, and finally a validated EV isolation and release methodology and kit using real samples.
The establishment of a total EV quantitation assay for the White Fox instrument able to quantitate the total EV
concentration in crude samples, i.e. in first instance applied to plasma and cell culture media.
The main achievements here are the EV quantitation assay and assay reagents, a specific EV quantitation analysis algorithm, and validation of EV quantification method using real samples from KOL.
Extend the current White FOx software capabilities to include features for supporting EV analysis applications, such as EV sample information, instrument controls for EV sample handling, real-time feedback on EV isolation and release, and EV quantitation. Tweak the software to support GLP and GMP users by upgrading the software to meet 21 CFR part 11 requirements. The main achievements are installers and all supporting documentation for these 2 software solutions.
The objective is to realize an effective go-to-market strategy to drive commercialisation of FOx EV applications, consumables and products. FOx will therefore develop a Marketing and Sales plan, based on further market assessment, including a Product Launch Readiness plan. FOx as an organisation also needs to scale-up operationally to support the commercialisation of FOx EV applications and products, for which an internal operational scale-up and HR strategy will be developed. The main achievements are a Design and Development Plan, including risk assessment/ mitigation plan & User Requirement specifications, an IP management and dissemination plan, and all other operational supporting and planning documentation.
FOX has established a breakthrough label-free technology in a fiber-optic concept in an automated platform that allows biomolecular interaction research of biologicals. The FOx affinity isolation and simultaneous quantification with a sensor dip probe is a new, selective (no bias), automated (scalable), faster and specific (purity >95%) method that allows accurately quantifying EV directly from crude samples. Moreover, it allows for the first time to release the quantified and specifically selected EV’s for further quantitative content analysis.
The main gain for the EV diagnostics developers and vendors is the ability to quantify EVs and additional protein
biomarkers directly in crude samples, such as blood samples. This enables them to use specific EV level biomarker whether in the membrane or content of the EV as diagnostic markers. FOx’s unique integrated EV isolation, quantification and characterization solution will significantly accelerate the introduction of new products (i.e. acceptance by authorities and the community will be facilitated based on undisputed results). An integrated EV solution combining high-quality EV isolation, quantitation, and characterization will improve (i.e.
timing/efficiency/reproducibility) the clinical development of EV-based diagnostics thus impacting the lives of many patients (e.g. cancer mortality reduction 20-25%) through earlier and personalized diagnosis and stratified treatment in cancer and cardiovascular disease areas.
The main gain for the EV diagnostics developers and vendors is the ability to quantify EVs and additional protein
biomarkers directly in crude samples, such as blood samples. This enables them to use specific EV level biomarker whether in the membrane or content of the EV as diagnostic markers. FOx’s unique integrated EV isolation, quantification and characterization solution will significantly accelerate the introduction of new products (i.e. acceptance by authorities and the community will be facilitated based on undisputed results). An integrated EV solution combining high-quality EV isolation, quantitation, and characterization will improve (i.e.
timing/efficiency/reproducibility) the clinical development of EV-based diagnostics thus impacting the lives of many patients (e.g. cancer mortality reduction 20-25%) through earlier and personalized diagnosis and stratified treatment in cancer and cardiovascular disease areas.