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The diagnosis and treatment of prostate cancer (PCa) is a major health-care concern worldwide. This cancer can manifest itself in many distinct forms and the transition from clinically indolent PCa to the more invasive aggressive form remains poorly understood. It is now universally accepted that glycan expression patterns change with the cellular modifications that accompany the onset of tumorigenesis. The aim of this study was to investigate if differential glycosylation patterns could distinguish between indolent, significant, and aggressive PCa. Whole serum N-glycan profiling was carried out on 117 prostate cancer patients’ serum using our automated, high-throughput analysis platform for glycan-profiling which utilizes ultra-performance liquid chromatography (UPLC) to obtain high resolution separation of N-linked glycans released from the serum glycoproteins. We observed increases in hybrid, oligomannose, and biantennary digalactosylated monosialylated glycans (M5A1G1S1, M8, and A2G2S1), bisecting glycans (A2B, A2(6)BG1) and monoantennary glycans (A1), and decreases in triantennary trigalactosylated trisialylated glycans with and without core fucose (A3G3S3 and FA3G3S3) with PCa progression from indolent through significant and aggressive disease. These changes give us an insight into the disease pathogenesis and identify potential biomarkers for monitoring the PCa progression, however these need further confirmation studies.

The diagnosis and treatment of prostate cancer (PCa) is a major health-care concern worldwide. This cancer can manifest itself in many distinct forms and the transition from clinically indolent PCa to the more invasive aggressive form remains poorly understood. It is now universally accepted that glycan expression patterns change with the cellular modifications that accompany the onset of tumorigenesis. The aim of this study was to investigate if differential glycosylation patterns could distinguish between indolent, significant, and aggressive PCa. Whole serum N-glycan profiling was carried out on 117 prostate cancer patients’ serum using our automated, high-throughput analysis platform for glycan-profiling which utilizes ultra-performance liquid chromatography (UPLC) to obtain high resolution separation of N-linked glycans released from the serum glycoproteins. We observed increases in hybrid, oligomannose, and biantennary digalactosylated monosialylated glycans (M5A1G1S1, M8, and A2G2S1), bisecting glycans (A2B, A2(6)BG1) and monoantennary glycans (A1), and decreases in triantennary trigalactosylated trisialylated glycans with and without core fucose (A3G3S3 and FA3G3S3) with PCa progression from indolent through significant and aggressive disease. These changes give us an insight into the disease pathogenesis and identify potential biomarkers for monitoring the PCa progression, however these need further confirmation studies.

Mycotoxins are toxic secondary fungal metabolites which contaminate a variety of food and feed products and pose serious health risks to humans and animals. Exposure to even low concentrations of these toxins can result in chronic diseases, with immunosuppressive and carcinogenic effects being of greatest concern. They are estimated to be found in 25% of the world’s food supply and have huge economic consequences globally. There is a lack of understanding of how mycotoxins interact with the immune system. The purpose of this research was to increase this understanding by examining the cytotoxic and immunomodulatory effects of the mycotoxins patulin, DON, ZEN and T-2 toxin in vitro, using the murine macrophage cell line, J774A.1. The results clearly demonstrated that T-2 toxin and ZEN significantly affect cell viability at high concentrations while sub-lethal concentrations of patulin, DON, ZEN and T-2 toxin, alone and in combinations, had a significant effect on host defence functions, through deregulation of IL-6, IL-10, IL-27, IL-12p40, IL-1β and TNF-α cytokine production, phagocytosis, and the cell surface expression of CD80, CD86, TLR4 and MHCII. The effects of these toxins are dependent on concentrations and combinations. This knowledge should be taken into consideration in the implementation of detection limits, aimed at minimising risks to human and animal health from mycotoxin exposure. A possible solution to this was explored through the use of a novel microfluidic technology, Lab-in-a-Trench, which demonstrated the ability to measure the immunomodulatory effects of mycotoxin combinations in ‘real-time’. Increased awareness of the hazards presented by mycotoxins has also led to the need for rapid detection systems. Recombinant antibodies incorporated into sensitive immunoassays offers the ability to satisfy this need. This thesis describes optimisation of immune library production for the mycotoxins patulin, ochratoxin A and aflatoxin M1 using an avian host

Mycotoxins are toxic secondary fungal metabolites which contaminate a variety of food and feed products and pose serious health risks to humans and animals. Exposure to even low concentrations of these toxins can result in chronic diseases, with immunosuppressive and carcinogenic effects being of greatest concern. They are estimated to be found in 25% of the world’s food supply and have huge economic consequences globally. There is a lack of understanding of how mycotoxins interact with the immune system. The purpose of this research was to increase this understanding by examining the cytotoxic and immunomodulatory effects of the mycotoxins patulin, DON, ZEN and T-2 toxin in vitro, using the murine macrophage cell line, J774A.1. The results clearly demonstrated that T-2 toxin and ZEN significantly affect cell viability at high concentrations while sub-lethal concentrations of patulin, DON, ZEN and T-2 toxin, alone and in combinations, had a significant effect on host defence functions, through deregulation of IL-6, IL-10, IL-27, IL-12p40, IL-1β and TNF-α cytokine production, phagocytosis, and the cell surface expression of CD80, CD86, TLR4 and MHCII. The effects of these toxins are dependent on concentrations and combinations. This knowledge should be taken into consideration in the implementation of detection limits, aimed at minimising risks to human and animal health from mycotoxin exposure. A possible solution to this was explored through the use of a novel microfluidic technology, Lab-in-a-Trench, which demonstrated the ability to measure the immunomodulatory effects of mycotoxin combinations in ‘real-time’. Increased awareness of the hazards presented by mycotoxins has also led to the need for rapid detection systems. Recombinant antibodies incorporated into sensitive immunoassays offers the ability to satisfy this need. This thesis describes optimisation of immune library production for the mycotoxins patulin, ochratoxin A and aflatoxin M1 using an avian host

Point-of-care diagnostics devices require user friendly automation of multi-step / multi-reagent bioanalytical protocols. We demonstrate, for the first time, Laser-actuated, Centrifugo-Pneumatic Valves (LCPVs) which open on-demand by ablating a remotely connected polymeric membrane with an instrument-based laser. Due to its small footprint, isolation of radiation-induced heating from the biological fluids and independence of radial position & contact-angle, the novel LCPV technology enables large–scale integration of flexibly customizable and operationally robust centrifugal microfluidic “Lab-on-a-Disc” (LoaD) systems. As a pilot application, we demonstrate the detection of Fatty Acid Binding Protein and Myeloperoxidase, both potential biomarkers used for cardiovascular disease (CVD) determination.

Point-of-care diagnostics devices require user friendly automation of multi-step / multi-reagent bioanalytical protocols. We demonstrate, for the first time, Laser-actuated, Centrifugo-Pneumatic Valves (LCPVs) which open on-demand by ablating a remotely connected polymeric membrane with an instrument-based laser. Due to its small footprint, isolation of radiation-induced heating from the biological fluids and independence of radial position & contact-angle, the novel LCPV technology enables large–scale integration of flexibly customizable and operationally robust centrifugal microfluidic “Lab-on-a-Disc” (LoaD) systems. As a pilot application, we demonstrate the detection of Fatty Acid Binding Protein and Myeloperoxidase, both potential biomarkers used for cardiovascular disease (CVD) determination.

—With the ubiquitous use of smartphones and other connected pieces of equipment, the number of devices connected to the internet is exponentially growing. This will test the efficiency of the envisioned 5G network architectures for data acquisition and its storage. It is a common observation that the communication between smart devices is typically influenced by their social relationship. This suggests that the theory of social networks can be leveraged to improve the quality of service for such communication links. In fact, the social networking concepts of centrality and community have been investigated for an efficient realization of novel wireless network architectures. This work provides a comprehensive introduction to social networks and reviews the recent literature on the application of social networks in wireless communications. The potential challenges in communication network design are also highlighted, for a successful implementation of social networking strategies. Finally, some future directions are discussed for the application of social networking strategies to emerging wireless technologies such as non-orthogonal multiple access and visible light communications.

Point-of-care diagnostics devices require user friendly automation of multi-step / multi-reagent bioanalytical protocols. We demonstrate, for the first time, Laser-actuated, Centrifugo-Pneumatic Valves (LCPVs) which open on-demand by ablating a remotely connected polymeric membrane with an instrument-based laser. Due to its small footprint, isolation of radiation-induced heating from the biological fluids and independence of radial position & contact-angle, the novel LCPV technology enables large–scale integration of flexibly customizable and operationally robust centrifugal microfluidic “Lab-on-a-Disc” (LoaD) systems. As a pilot application, we demonstrate the detection of Fatty Acid Binding Protein and Myeloperoxidase, both potential biomarkers used for cardiovascular disease (CVD) determination.

A multi-marker approach is increasingly considered in diagnosis of prostate cancer to minimize the number of unnecessary biopsies [1]. Centrifugo-pneumatic flow control is often the method of choice for multiplexing such assays [2]. However, the number of assays that can be parallelized on such a dissolvable-film (DF) based system is limited by the substantial spread of the burst frequencies [2] owing to manufacturing tolerances. In this work we substantially enhance the definition of actuation frequencies by 75% using a novel design to enable the automation of multi-step / multi-reagent Lab-on-a-Disc (LoaD) platform. We demonstrate a multi-marker prostate cancer immunoassay panel.

Cyclic peptide cyanobacterial toxins, in particular Microcystis aeruginosa, pose a serious health risk to humans and animals alike [1], [2]. Occurring mostly in fresh and brackish water, they have been identified to cause cancer promotion and liver damage [3]. Herein, we describe a portable, microfluidic-based system for in-situ detection of algal toxins in fresh water. The technology development presented here is a fully integrated and portable sample-to-answer centrifugal microfluidics-based system for the detection of toxic cyanobacteria – Microcystin-LR in fresh water. Our unique system employs highly-specific recombinant chicken anti-microcystin antibodies, prepared in-house, with a 3D-printed ‘LASER-photo¬diode’ fluorescent detection technique, also developed in-house. The system has high analytical specificity and sensitivity for detection of toxins below the regulatory limit with intra/inter day coefficient of variation of less than 20%. Dissolvable-film based valving technique was used for flow actuation and integration of multiple assays on the centrifugal cartridge. This new approach forms the basis of a cost efficient, USB-controlled water quality monitoring system. Technically, this integrated system consists of two components; a microfluidic disc (figure 1.A), the disc-holder fabricated and assembled from a 3D-printed casing, with electronic components housed in device. The 5-layered microfluidic disc consists of five reservoirs (figure 1.B), each with a separate venti-lation, aligned radially with inter-connected microchannels. A competitive immunoassay format is utilised to detect free toxin (figure 1.C). Sensitivity, reproducibility and ease-of-use are key features of this monitoring device. The ‘top-down’ optical detection system has been modified for improved detection sensitivity, as well as the elimination of external noise.

A multi-marker approach is increasingly considered in diagnosis of prostate cancer to minimize the number of unnecessary biopsies [1]. Centrifugo-pneumatic flow control is often the method of choice for multiplexing such assays [2]. However, the number of assays that can be parallelized on such a dissolvable-film (DF) based system is limited by the substantial spread of the burst frequencies [2] owing to manufacturing tolerances. In this work we substantially enhance the definition of actuation frequencies by 75% using a novel design to enable the automation of multi-step / multi-reagent Lab-on-a-Disc (LoaD) platform. We demonstrate a multi-marker prostate cancer immunoassay panel.

Cyclic peptide cyanobacterial toxins, in particular Microcystis aeruginosa, pose a serious health risk to humans and animals alike [1], [2]. Occurring mostly in fresh and brackish water, they have been identified to cause cancer promotion and liver damage [3]. Herein, we describe a portable, microfluidic-based system for in-situ detection of algal toxins in fresh water. The technology development presented here is a fully integrated and portable sample-to-answer centrifugal microfluidics-based system for the detection of toxic cyanobacteria – Microcystin-LR in fresh water. Our unique system employs highly-specific recombinant chicken anti-microcystin antibodies, prepared in-house, with a 3D-printed ‘LASER-photo¬diode’ fluorescent detection technique, also developed in-house. The system has high analytical specificity and sensitivity for detection of toxins below the regulatory limit with intra/inter day coefficient of variation of less than 20%. Dissolvable-film based valving technique was used for flow actuation and integration of multiple assays on the centrifugal cartridge. This new approach forms the basis of a cost efficient, USB-controlled water quality monitoring system. Technically, this integrated system consists of two components; a microfluidic disc (figure 1.A), the disc-holder fabricated and assembled from a 3D-printed casing, with electronic components housed in device. The 5-layered microfluidic disc consists of five reservoirs (figure 1.B), each with a separate venti-lation, aligned radially with inter-connected microchannels. A competitive immunoassay format is utilised to detect free toxin (figure 1.C). Sensitivity, reproducibility and ease-of-use are key features of this monitoring device. The ‘top-down’ optical detection system has been modified for improved detection sensitivity, as well as the elimination of external noise.

A multi-marker approach is increasingly considered in diagnosis of prostate cancer to minimize the number of unnecessary biopsies [1]. Centrifugo-pneumatic flow control is often the method of choice for multiplexing such assays [2]. However, the number of assays that can be parallelized on such a dissolvable-film (DF) based system is limited by the substantial spread of the burst frequencies [2] owing to manufacturing tolerances. In this work we substantially enhance the definition of actuation frequencies by 75% using a novel design to enable the automation of multi-step / multi-reagent Lab-on-a-Disc (LoaD) platform. We demonstrate a multi-marker prostate cancer immunoassay panel.

Caroline Murphy, Tatyana Devine, Richard O'Kennedy School of Biotechnology and Biomedical Diagnostics Institute, Dublin City University, Glasnevin, Dublin, Ireland Abstract: The separation of antibodies from complex mixtures can be achieved using chromatographic or non-chromatographic techniques. The purification of antibodies using chromatography involves the separation of antibodies or antibody-derived molecules present in complex mixtures by passing them through a solid phase (eg, silica resin or beads, monolithic columns, or cellulose membranes) and allowing the antibodies to bind or pass through depending on whether "bind-and-elute" or "flow-through" chromatographic methods are employed. Chromatographic methodologies can incorporate different separation techniques, such as affinity-tag binding, ion-exchange, size-exclusion chromatography, or immunoaffinity chromatography. However, in a concerted effort to become less reliant on chromatography-based separations owing to the high cost of large-scale production, non-chromatographic-based techniques such as precipitation, flocculation, crystallization, filtration, and aqueous two-phase partitioning are now becoming more popular. This review details current chromatographic and non-chromatographic methodologies used for the purification of antibodies and expands on technological advancements and practical uses that have recently been reported. Keywords: antibody, chromatography-based purification, technology advancements, non-chromatographic purification, advances in purification technology, separation techniques Corrigendum for this paper has been published&nbsp

A multi-marker approach is increasingly considered in diagnosis of prostate cancer to minimize the number of unnecessary biopsies [1]. Centrifugo-pneumatic flow control is often the method of choice for multiplexing such assays [2]. However, the number of assays that can be parallelized on such a dissolvable-film (DF) based system is limited by the substantial spread of the burst frequencies [2] owing to manufacturing tolerances. In this work we substantially enhance the definition of actuation frequencies by 75% using a novel design to enable the automation of multi-step / multi-reagent Lab-on-a-Disc (LoaD) platform. We demonstrate a multi-marker prostate cancer immunoassay panel.

Cyclic peptide cyanobacterial toxins, in particular Microcystis aeruginosa, pose a serious health risk to humans and animals alike [1], [2]. Occurring mostly in fresh and brackish water, they have been identified to cause cancer promotion and liver damage [3]. Herein, we describe a portable, microfluidic-based system for in-situ detection of algal toxins in fresh water. The technology development presented here is a fully integrated and portable sample-to-answer centrifugal microfluidics-based system for the detection of toxic cyanobacteria – Microcystin-LR in fresh water. Our unique system employs highly-specific recombinant chicken anti-microcystin antibodies, prepared in-house, with a 3D-printed ‘LASER-photo¬diode’ fluorescent detection technique, also developed in-house. The system has high analytical specificity and sensitivity for detection of toxins below the regulatory limit with intra/inter day coefficient of variation of less than 20%. Dissolvable-film based valving technique was used for flow actuation and integration of multiple assays on the centrifugal cartridge. This new approach forms the basis of a cost efficient, USB-controlled water quality monitoring system. Technically, this integrated system consists of two components; a microfluidic disc (figure 1.A), the disc-holder fabricated and assembled from a 3D-printed casing, with electronic components housed in device. The 5-layered microfluidic disc consists of five reservoirs (figure 1.B), each with a separate venti-lation, aligned radially with inter-connected microchannels. A competitive immunoassay format is utilised to detect free toxin (figure 1.C). Sensitivity, reproducibility and ease-of-use are key features of this monitoring device. The ‘top-down’ optical detection system has been modified for improved detection sensitivity, as well as the elimination of external noise.

Globally, the most prevalent cyanobacterial toxins, in blooms from fresh and brackish waters, are the cyclic peptide toxins of the microcystin family. The need for on-site algal-toxin monitoring has become increasingly urgent due to the amplified demand for fresh-water and for safe, ‘toxinfree’ shellfish and fish stocks. Along with routine testing of shellfish stocks for biotoxins including microcystin, the EU also require routine monitoring for the presence of the causative algal species. Herein, we describe a novel, Lab-On-A-Disc (LOAD) platform which has been developed to assess microcystin toxin levels in-situ. Using recombinant antibody technology, the LOAD platform combines immunofluorescence with centrifugally driven microfluidic liquid handling to achieve a next-generation disposable device for high throughput sampling. A low-complexity, ‘LED-Photodiode’ based optical sensing system was tailor-made for the system, which allows the fluorescence signal of the microcystin-specific reaction to be quantified. This system can rapidly and accurately detect the presence of microcystin in less than 10 minutes with minimum user interaction and maximum reproducibility. This method provides a low cost diagnostic alternative to the current laborious and costly methods used for toxin monitoring.

Cyclic peptide cyanobacterial toxins, in particular Microcystis aeruginosa, pose a serious health risk to humans and animals alike [1], [2]. Occurring mostly in fresh and brackish water, they have been identified to cause cancer promotion and liver damage [3]. Herein, we describe a portable, microfluidic-based system for in-situ detection of algal toxins in fresh water. The Atalanta system is a novel, portable and sample-to-answer platform for the detection of toxic cyanobacteria – Microcystin-LR in fresh water. Atalanta utilises the partnership of highly-specific recombinant chicken anti-microcystin antibodies, prepared in-house, with a 3D-printed ‘LASER-photo¬diode’ fluorescent detection method, also developed in-house. A competitive immunoassay format is utilised to detect free toxin. Furthermore, dissolvable-film (DF) based flow-actuation facilitates full assay inte¬gration. This new approach will form the basis of a cost efficient, USB-controlled water quality monitoring system. The Atalanta detection system consists of two components; the microfluidic Atalanta disc and the disc-holder. The Atalanta disc-holder was fabricated and assembled from a 3D-printed casing, with electronic components housed in device. The 5-layered microfluidic disc consists of five reservoirs, each with a separate venti¬lation, aligned radially with inter-connected microchannels. Each reservoir represents a functional assay step. First, microcystin conjugate is coated to the functionalised surface of the reservoir 3 prior to assembling the disc. A freshwater sample in reservoir 1 is pre-incubated with recombinant antibodies labelled with fluorophore (Alexa 647) in reservoir 2. This is then spun into reservoir 3 for detection through a competitive immunoassay using Microcystin-LR. Low fluorescence signal indicates high Microcystin-LR concentration in the sample.

Globally, the most prevalent cyanobacterial toxins, in blooms from fresh and brackish waters, are the cyclic peptide toxins of the microcystin family. The need for on-site algal-toxin monitoring has become increasingly urgent due to the amplified demand for fresh-water and for safe, ‘toxinfree’ shellfish and fish stocks. Along with routine testing of shellfish stocks for biotoxins including microcystin, the EU also require routine monitoring for the presence of the causative algal species. Herein, we describe a novel, Lab-On-A-Disc (LOAD) platform which has been developed to assess microcystin toxin levels in-situ. Using recombinant antibody technology, the LOAD platform combines immunofluorescence with centrifugally driven microfluidic liquid handling to achieve a next-generation disposable device for high throughput sampling. A low-complexity, ‘LED-Photodiode’ based optical sensing system was tailor-made for the system, which allows the fluorescence signal of the microcystin-specific reaction to be quantified. This system can rapidly and accurately detect the presence of microcystin in less than 10 minutes with minimum user interaction and maximum reproducibility. This method provides a low cost diagnostic alternative to the current laborious and costly methods used for toxin monitoring.

Globally, the most prevalent cyanobacterial toxins, in blooms from fresh and brackish waters, are the cyclic peptide toxins of the microcystin family. The need for on-site algal-toxin monitoring has become increasingly urgent due to the amplified demand for fresh-water and for safe, ‘toxin-free’ shellfish and fish stocks. Along with routine testing of shellfish stocks for biotoxins including microcystin, the EU also require routine monitoring for the presence of the causative algal species. Herein, we describe a novel, Lab-On-A-Disc (LOAD) platform which has been developed to assess microcystin toxin levels in-situ. Using recombinant antibody technology, the LOAD platform combines immunofluorescence with centrifugally driven microfluidic liquid handling to achieve a next-generation disposable device for high throughput sampling. A low-complexity, ‘LED-Photodiode’ based optical sensing system was tailor-made for the system, which allows the fluorescence signal of the microcystin-specific reaction to be quantified. This system can rapidly and accurately detect the presence of microcystin in less than 10 minutes with minimum user interaction and maximum reproducibility. This method provides a low cost diagnostic alternative to the current laborious and costly methods used for toxin monitoring.

With population growth, there is an ever-increasing requirement for fresh-water sources and safe, ‘toxin-free’ shellfish and fish stocks. To monitor these resources carefully, there is a need for the development of an in-situ algal-toxin monitoring solutions. Presented here is one of these solutions; The ToxiSense Detection system (Figure 1). This system has been designed to be as generically applicable to toxin detection as possible, with the cyanobacterial Microcystin-LR used for proof-of-concept.

Cyclic peptide cyanobacterial toxins, in particular Microcystis aeruginosa, pose a serious health risk to humans and animals alike [1], [2]. Occurring mostly in fresh and brackish water, they have been identified to cause cancer promotion and liver damage [3]. Herein, we describe a portable, microfluidic-based system for in-situ detection of algal toxins in fresh water. The Atalanta system is a novel, portable and sample-to-answer platform for the detection of toxic cyanobacteria – Microcystin-LR in fresh water. Atalanta utilises the partnership of highly-specific recombinant chicken anti-microcystin antibodies, prepared in-house, with a 3D-printed ‘LASER-photo¬diode’ fluorescent detection method, also developed in-house. A competitive immunoassay format is utilised to detect free toxin. Furthermore, dissolvable-film (DF) based flow-actuation facilitates full assay inte¬gration. This new approach will form the basis of a cost efficient, USB-controlled water quality monitoring system. The Atalanta detection system consists of two components; the microfluidic Atalanta disc and the disc-holder. The Atalanta disc-holder was fabricated and assembled from a 3D-printed casing, with electronic components housed in device. The 5-layered microfluidic disc consists of five reservoirs, each with a separate venti¬lation, aligned radially with inter-connected microchannels. Each reservoir represents a functional assay step. First, microcystin conjugate is coated to the functionalised surface of the reservoir 3 prior to assembling the disc. A freshwater sample in reservoir 1 is pre-incubated with recombinant antibodies labelled with fluorophore (Alexa 647) in reservoir 2. This is then spun into reservoir 3 for detection through a competitive immunoassay using Microcystin-LR. Low fluorescence signal indicates high Microcystin-LR concentration in the sample.

Cyclic peptide cyanobacterial toxins, in particular Microcystis aeruginosa, pose a serious health risk to humans and animals alike [1], [2]. Occurring mostly in fresh and brackish water, they have been identified to cause cancer promotion and liver damage [3]. Herein, we describe a portable, microfluidic-based system for in-situ detection of algal toxins in fresh water. The technology development presented here is a fully integrated and portable sample-to-answer centrifugal microfluidics-based system for the detection of toxic cyanobacteria – Microcystin-LR in fresh water. Our unique system employs highly-specific recombinant chicken anti-microcystin antibodies, prepared in-house, with a 3D-printed ‘LASER-photo¬diode’ fluorescent detection technique, also developed in-house. The system has high analytical specificity and sensitivity for detection of toxins below the regulatory limit with intra/inter day coefficient of variation of less than 20%. Dissolvable-film based valving technique was used for flow actuation and integration of multiple assays on the centrifugal cartridge. This new approach forms the basis of a cost efficient, USB-controlled water quality monitoring system. Technically, this integrated system consists of two components; a microfluidic disc (figure 1.A), the disc-holder fabricated and assembled from a 3D-printed casing, with electronic components housed in device. The 5-layered microfluidic disc consists of five reservoirs (figure 1.B), each with a separate venti-lation, aligned radially with inter-connected microchannels. A competitive immunoassay format is utilised to detect free toxin (figure 1.C). Sensitivity, reproducibility and ease-of-use are key features of this monitoring device. The ‘top-down’ optical detection system has been modified for improved detection sensitivity, as well as the elimination of external noise.

Globally, the most prevalent cyanobacterial toxins, in blooms from fresh and brackish waters, are the cyclic peptide toxins of the microcystin family. The need for on-site algal-toxin monitoring has become increasingly urgent due to the amplified demand for fresh-water and for safe, ‘toxinfree’ shellfish and fish stocks. Along with routine testing of shellfish stocks for biotoxins including microcystin, the EU also require routine monitoring for the presence of the causative algal species. Herein, we describe a novel, Lab-On-A-Disc (LOAD) platform which has been developed to assess microcystin toxin levels in-situ. Using recombinant antibody technology, the LOAD platform combines immunofluorescence with centrifugally driven microfluidic liquid handling to achieve a next-generation disposable device for high throughput sampling. A low-complexity, ‘LED-Photodiode’ based optical sensing system was tailor-made for the system, which allows the fluorescence signal of the microcystin-specific reaction to be quantified. This system can rapidly and accurately detect the presence of microcystin in less than 10 minutes with minimum user interaction and maximum reproducibility. This method provides a low cost diagnostic alternative to the current laborious and costly methods used for toxin monitoring.

Globally, the most prevalent cyanobacterial toxins, in blooms from fresh and brackish waters, are the cyclic peptide toxins of the microcystin family. The need for on-site algal-toxin monitoring has become increasingly urgent due to the amplified demand for fresh-water and for safe, ‘toxin-free’ shellfish and fish stocks. Along with routine testing of shellfish stocks for biotoxins including microcystin, the EU also require routine monitoring for the presence of the causative algal species. Herein, we describe a novel, Lab-On-A-Disc (LOAD) platform which has been developed to assess microcystin toxin levels in-situ. Using recombinant antibody technology, the LOAD platform combines immunofluorescence with centrifugally driven microfluidic liquid handling to achieve a next-generation disposable device for high throughput sampling. A low-complexity, ‘LED-Photodiode’ based optical sensing system was tailor-made for the system, which allows the fluorescence signal of the microcystin-specific reaction to be quantified. This system can rapidly and accurately detect the presence of microcystin in less than 10 minutes with minimum user interaction and maximum reproducibility. This method provides a low cost diagnostic alternative to the current laborious and costly methods used for toxin monitoring.

Globally, the most prevalent cyanobacterial toxins, in blooms from fresh and brackish waters, are the cyclic peptide toxins of the microcystin family. The need for on-site algal-toxin monitoring has become increasingly urgent due to the amplified demand for fresh-water and for safe, ‘toxinfree’ shellfish and fish stocks. Along with routine testing of shellfish stocks for biotoxins including microcystin, the EU also require routine monitoring for the presence of the causative algal species. Herein, we describe a novel, Lab-On-A-Disc (LOAD) platform which has been developed to assess microcystin toxin levels in-situ. Using recombinant antibody technology, the LOAD platform combines immunofluorescence with centrifugally driven microfluidic liquid handling to achieve a next-generation disposable device for high throughput sampling. A low-complexity, ‘LED-Photodiode’ based optical sensing system was tailor-made for the system, which allows the fluorescence signal of the microcystin-specific reaction to be quantified. This system can rapidly and accurately detect the presence of microcystin in less than 10 minutes with minimum user interaction and maximum reproducibility. This method provides a low cost diagnostic alternative to the current laborious and costly methods used for toxin monitoring.

STRA6 is a plasma membrane protein that mediates the transport of vitamin A, or retinol, from plasma retinol binding protein (RBP) into the cell. Mutations in human STRA6 are associated with Matthew-Wood syndrome, which is characterized by severe developmental defects. Despite the obvious importance of this protein to human health, little is known about its structure and mechanism of action. To overcome the difficulties frequently encountered with the production of membrane proteins for structural determination, STRA6 has been expressed in Pichia pastoris as a fusion to green fluorescent protein (GFP), a strategy which has been a critical first step in solving the crystal structures of several membrane proteins. STRA6-GFP was correctly targeted to the cell surface where it bound RBP. Here we report the large-scale expression, purification and characterisation of STRA6GFP. One litre of culture, corresponding to 175 g cells, yielded about 1.5 mg of pure protein. The interaction between purified STRA6 and its ligand RBP was studied by surface plasmon resonance-based binding analysis. The interaction between STRA6 and RBP was not retinol-dependent and the binding data were consistent with a transient interaction of 1 mole RBP/mole STRA6.

STRA6 is a plasma membrane protein that mediates the transport of vitamin A, or retinol, from plasma retinol binding protein (RBP) into the cell. Mutations in human STRA6 are associated with Matthew-Wood syndrome, which is characterized by severe developmental defects. Despite the obvious importance of this protein to human health, little is known about its structure and mechanism of action. To overcome the difficulties frequently encountered with the production of membrane proteins for structural determination, STRA6 has been expressed in Pichia pastoris as a fusion to green fluorescent protein (GFP), a strategy which has been a critical first step in solving the crystal structures of several membrane proteins. STRA6-GFP was correctly targeted to the cell surface where it bound RBP. Here we report the large-scale expression, purification and characterisation of STRA6GFP. One litre of culture, corresponding to 175 g cells, yielded about 1.5 mg of pure protein. The interaction between purified STRA6 and its ligand RBP was studied by surface plasmon resonance-based binding analysis. The interaction between STRA6 and RBP was not retinol-dependent and the binding data were consistent with a transient interaction of 1 mole RBP/mole STRA6.

Globally, the most prevalent cyanobacterial toxins, in blooms from fresh and brackish waters, are the cyclic peptide toxins of the microcystin family. The need for on-site algal-toxin monitoring has become increasingly urgent due to the amplified demand for fresh-water and for safe, ‘toxin-free’ shellfish and fish stocks. Along with routine testing of shellfish stocks for biotoxins including microcystin, the EU also require routine monitoring for the presence of the causative algal species. Herein, we describe a novel, Lab-On-A-Disc (LOAD) platform which has been developed to assess microcystin toxin levels in-situ. Using recombinant antibody technology, the LOAD platform combines immunofluorescence with centrifugally driven microfluidic liquid handling to achieve a next-generation disposable device for high throughput sampling. A low-complexity, ‘LED-Photodiode’ based optical sensing system was tailor-made for the system, which allows the fluorescence signal of the microcystin-specific reaction to be quantified. This system can rapidly and accurately detect the presence of microcystin in less than 10 minutes with minimum user interaction and maximum reproducibility. This method provides a low cost diagnostic alternative to the current laborious and costly methods used for toxin monitoring.

Cyclic peptide cyanobacterial toxins, in particular Microcystis aeruginosa, pose a serious health risk to humans and animals alike [1], [2]. Occurring mostly in fresh and brackish water, they have been identified to cause cancer promotion and liver damage [3]. Herein, we describe a portable, microfluidic-based system for in-situ detection of algal toxins in fresh water. The Atalanta system is a novel, portable and sample-to-answer platform for the detection of toxic cyanobacteria – Microcystin-LR in fresh water. Atalanta utilises the partnership of highly-specific recombinant chicken anti-microcystin antibodies, prepared in-house, with a 3D-printed ‘LASER-photo¬diode’ fluorescent detection method, also developed in-house. A competitive immunoassay format is utilised to detect free toxin. Furthermore, dissolvable-film (DF) based flow-actuation facilitates full assay inte¬gration. This new approach will form the basis of a cost efficient, USB-controlled water quality monitoring system. The Atalanta detection system consists of two components; the microfluidic Atalanta disc and the disc-holder. The Atalanta disc-holder was fabricated and assembled from a 3D-printed casing, with electronic components housed in device. The 5-layered microfluidic disc consists of five reservoirs, each with a separate venti¬lation, aligned radially with inter-connected microchannels. Each reservoir represents a functional assay step. First, microcystin conjugate is coated to the functionalised surface of the reservoir 3 prior to assembling the disc. A freshwater sample in reservoir 1 is pre-incubated with recombinant antibodies labelled with fluorophore (Alexa 647) in reservoir 2. This is then spun into reservoir 3 for detection through a competitive immunoassay using Microcystin-LR. Low fluorescence signal indicates high Microcystin-LR concentration in the sample.

Cyclic peptide cyanobacterial toxins, in particular Microcystis aeruginosa, pose a serious health risk to humans and animals alike [1], [2]. Occurring mostly in fresh and brackish water, they have been identified to cause cancer promotion and liver damage [3]. Herein, we describe a portable, microfluidic-based system for in-situ detection of algal toxins in fresh water. The Atalanta system is a novel, portable and sample-to-answer platform for the detection of toxic cyanobacteria – Microcystin-LR in fresh water. Atalanta utilises the partnership of highly-specific recombinant chicken anti-microcystin antibodies, prepared in-house, with a 3D-printed ‘LASER-photo¬diode’ fluorescent detection method, also developed in-house. A competitive immunoassay format is utilised to detect free toxin. Furthermore, dissolvable-film (DF) based flow-actuation facilitates full assay inte¬gration. This new approach will form the basis of a cost efficient, USB-controlled water quality monitoring system. The Atalanta detection system consists of two components; the microfluidic Atalanta disc and the disc-holder. The Atalanta disc-holder was fabricated and assembled from a 3D-printed casing, with electronic components housed in device. The 5-layered microfluidic disc consists of five reservoirs, each with a separate venti¬lation, aligned radially with inter-connected microchannels. Each reservoir represents a functional assay step. First, microcystin conjugate is coated to the functionalised surface of the reservoir 3 prior to assembling the disc. A freshwater sample in reservoir 1 is pre-incubated with recombinant antibodies labelled with fluorophore (Alexa 647) in reservoir 2. This is then spun into reservoir 3 for detection through a competitive immunoassay using Microcystin-LR. Low fluorescence signal indicates high Microcystin-LR concentration in the sample.

Globally, the most prevalent cyanobacterial toxins, in blooms from fresh and brackish waters, are the cyclic peptide toxins of the microcystin family. The need for on-site algal-toxin monitoring has become increasingly urgent due to the amplified demand for fresh-water and for safe, ‘toxin-free’ shellfish and fish stocks. Along with routine testing of shellfish stocks for biotoxins including microcystin, the EU also require routine monitoring for the presence of the causative algal species. Herein, we describe a novel, Lab-On-A-Disc (LOAD) platform which has been developed to assess microcystin toxin levels in-situ. Using recombinant antibody technology, the LOAD platform combines immunofluorescence with centrifugally driven microfluidic liquid handling to achieve a next-generation disposable device for high throughput sampling. A low-complexity, ‘LED-Photodiode’ based optical sensing system was tailor-made for the system, which allows the fluorescence signal of the microcystin-specific reaction to be quantified. This system can rapidly and accurately detect the presence of microcystin in less than 10 minutes with minimum user interaction and maximum reproducibility. This method provides a low cost diagnostic alternative to the current laborious and costly methods used for toxin monitoring.

Cyclic peptide cyanobacterial toxins, in particular Microcystis aeruginosa, pose a serious health risk to humans and animals alike [1], [2]. Occurring mostly in fresh and brackish water, they have been identified to cause cancer promotion and liver damage [3]. Herein, we describe a portable, microfluidic-based system for in-situ detection of algal toxins in fresh water. The technology development presented here is a fully integrated and portable sample-to-answer centrifugal microfluidics-based system for the detection of toxic cyanobacteria – Microcystin-LR in fresh water. Our unique system employs highly-specific recombinant chicken anti-microcystin antibodies, prepared in-house, with a 3D-printed ‘LASER-photo¬diode’ fluorescent detection technique, also developed in-house. The system has high analytical specificity and sensitivity for detection of toxins below the regulatory limit with intra/inter day coefficient of variation of less than 20%. Dissolvable-film based valving technique was used for flow actuation and integration of multiple assays on the centrifugal cartridge. This new approach forms the basis of a cost efficient, USB-controlled water quality monitoring system. Technically, this integrated system consists of two components; a microfluidic disc (figure 1.A), the disc-holder fabricated and assembled from a 3D-printed casing, with electronic components housed in device. The 5-layered microfluidic disc consists of five reservoirs (figure 1.B), each with a separate venti-lation, aligned radially with inter-connected microchannels. A competitive immunoassay format is utilised to detect free toxin (figure 1.C). Sensitivity, reproducibility and ease-of-use are key features of this monitoring device. The ‘top-down’ optical detection system has been modified for improved detection sensitivity, as well as the elimination of external noise.

Cyclic peptide cyanobacterial toxins, in particular Microcystis aeruginosa, pose a serious health risk to humans and animals alike [1], [2]. Occurring mostly in fresh and brackish water, they have been identified to cause cancer promotion and liver damage [3]. Herein, we describe a portable, microfluidic-based system for in-situ detection of algal toxins in fresh water. The Atalanta system is a novel, portable and sample-to-answer platform for the detection of toxic cyanobacteria – Microcystin-LR in fresh water. Atalanta utilises the partnership of highly-specific recombinant chicken anti-microcystin antibodies, prepared in-house, with a 3D-printed ‘LASER-photo¬diode’ fluorescent detection method, also developed in-house. A competitive immunoassay format is utilised to detect free toxin. Furthermore, dissolvable-film (DF) based flow-actuation facilitates full assay inte¬gration. This new approach will form the basis of a cost efficient, USB-controlled water quality monitoring system. The Atalanta detection system consists of two components; the microfluidic Atalanta disc and the disc-holder. The Atalanta disc-holder was fabricated and assembled from a 3D-printed casing, with electronic components housed in device. The 5-layered microfluidic disc consists of five reservoirs, each with a separate venti¬lation, aligned radially with inter-connected microchannels. Each reservoir represents a functional assay step. First, microcystin conjugate is coated to the functionalised surface of the reservoir 3 prior to assembling the disc. A freshwater sample in reservoir 1 is pre-incubated with recombinant antibodies labelled with fluorophore (Alexa 647) in reservoir 2. This is then spun into reservoir 3 for detection through a competitive immunoassay using Microcystin-LR. Low fluorescence signal indicates high Microcystin-LR concentration in the sample.

With population growth, there is an ever-increasing requirement for fresh-water sources and safe, ‘toxin-free’ shellfish and fish stocks. To monitor these resources carefully, there is a need for the development of an in-situ algal-toxin monitoring solutions. Presented here is one of these solutions; The ToxiSense Detection system (Figure 1). This system has been designed to be as generically applicable to toxin detection as possible, with the cyanobacterial Microcystin-LR used for proof-of-concept.

Cyclic peptide cyanobacterial toxins, in particular Microcystis aeruginosa, pose a serious health risk to humans and animals alike [1], [2]. Occurring mostly in fresh and brackish water, they have been identified to cause cancer promotion and liver damage [3]. Herein, we describe a portable, microfluidic-based system for in-situ detection of algal toxins in fresh water. The technology development presented here is a fully integrated and portable sample-to-answer centrifugal microfluidics-based system for the detection of toxic cyanobacteria – Microcystin-LR in fresh water. Our unique system employs highly-specific recombinant chicken anti-microcystin antibodies, prepared in-house, with a 3D-printed ‘LASER-photo¬diode’ fluorescent detection technique, also developed in-house. The system has high analytical specificity and sensitivity for detection of toxins below the regulatory limit with intra/inter day coefficient of variation of less than 20%. Dissolvable-film based valving technique was used for flow actuation and integration of multiple assays on the centrifugal cartridge. This new approach forms the basis of a cost efficient, USB-controlled water quality monitoring system. Technically, this integrated system consists of two components; a microfluidic disc (figure 1.A), the disc-holder fabricated and assembled from a 3D-printed casing, with electronic components housed in device. The 5-layered microfluidic disc consists of five reservoirs (figure 1.B), each with a separate venti-lation, aligned radially with inter-connected microchannels. A competitive immunoassay format is utilised to detect free toxin (figure 1.C). Sensitivity, reproducibility and ease-of-use are key features of this monitoring device. The ‘top-down’ optical detection system has been modified for improved detection sensitivity, as well as the elimination of external noise.

Globally, the most prevalent cyanobacterial toxins, in blooms from fresh and brackish waters, are the cyclic peptide toxins of the microcystin family. The need for on-site algal-toxin monitoring has become increasingly urgent due to the amplified demand for fresh-water and for safe, ‘toxinfree’ shellfish and fish stocks. Along with routine testing of shellfish stocks for biotoxins including microcystin, the EU also require routine monitoring for the presence of the causative algal species. Herein, we describe a novel, Lab-On-A-Disc (LOAD) platform which has been developed to assess microcystin toxin levels in-situ. Using recombinant antibody technology, the LOAD platform combines immunofluorescence with centrifugally driven microfluidic liquid handling to achieve a next-generation disposable device for high throughput sampling. A low-complexity, ‘LED-Photodiode’ based optical sensing system was tailor-made for the system, which allows the fluorescence signal of the microcystin-specific reaction to be quantified. This system can rapidly and accurately detect the presence of microcystin in less than 10 minutes with minimum user interaction and maximum reproducibility. This method provides a low cost diagnostic alternative to the current laborious and costly methods used for toxin monitoring.

Cyclic peptide cyanobacterial toxins, in particular Microcystis aeruginosa, pose a serious health risk to humans and animals alike [1], [2]. Occurring mostly in fresh and brackish water, they have been identified to cause cancer promotion and liver damage [3]. Herein, we describe a portable, microfluidic-based system for in-situ detection of algal toxins in fresh water. The Atalanta system is a novel, portable and sample-to-answer platform for the detection of toxic cyanobacteria – Microcystin-LR in fresh water. Atalanta utilises the partnership of highly-specific recombinant chicken anti-microcystin antibodies, prepared in-house, with a 3D-printed ‘LASER-photo¬diode’ fluorescent detection method, also developed in-house. A competitive immunoassay format is utilised to detect free toxin. Furthermore, dissolvable-film (DF) based flow-actuation facilitates full assay inte¬gration. This new approach will form the basis of a cost efficient, USB-controlled water quality monitoring system. The Atalanta detection system consists of two components; the microfluidic Atalanta disc and the disc-holder. The Atalanta disc-holder was fabricated and assembled from a 3D-printed casing, with electronic components housed in device. The 5-layered microfluidic disc consists of five reservoirs, each with a separate venti¬lation, aligned radially with inter-connected microchannels. Each reservoir represents a functional assay step. First, microcystin conjugate is coated to the functionalised surface of the reservoir 3 prior to assembling the disc. A freshwater sample in reservoir 1 is pre-incubated with recombinant antibodies labelled with fluorophore (Alexa 647) in reservoir 2. This is then spun into reservoir 3 for detection through a competitive immunoassay using Microcystin-LR. Low fluorescence signal indicates high Microcystin-LR concentration in the sample.

Cyclic peptide cyanobacterial toxins, in particular Microcystis aeruginosa, pose a serious health risk to humans and animals alike [1], [2]. Occurring mostly in fresh and brackish water, they have been identified to cause cancer promotion and liver damage [3]. Herein, we describe a portable, microfluidic-based system for in-situ detection of algal toxins in fresh water. The Atalanta system is a novel, portable and sample-to-answer platform for the detection of toxic cyanobacteria – Microcystin-LR in fresh water. Atalanta utilises the partnership of highly-specific recombinant chicken anti-microcystin antibodies, prepared in-house, with a 3D-printed ‘LASER-photo¬diode’ fluorescent detection method, also developed in-house. A competitive immunoassay format is utilised to detect free toxin. Furthermore, dissolvable-film (DF) based flow-actuation facilitates full assay inte¬gration. This new approach will form the basis of a cost efficient, USB-controlled water quality monitoring system. The Atalanta detection system consists of two components; the microfluidic Atalanta disc and the disc-holder. The Atalanta disc-holder was fabricated and assembled from a 3D-printed casing, with electronic components housed in device. The 5-layered microfluidic disc consists of five reservoirs, each with a separate venti¬lation, aligned radially with inter-connected microchannels. Each reservoir represents a functional assay step. First, microcystin conjugate is coated to the functionalised surface of the reservoir 3 prior to assembling the disc. A freshwater sample in reservoir 1 is pre-incubated with recombinant antibodies labelled with fluorophore (Alexa 647) in reservoir 2. This is then spun into reservoir 3 for detection through a competitive immunoassay using Microcystin-LR. Low fluorescence signal indicates high Microcystin-LR concentration in the sample.

With population growth, there is an ever-increasing requirement for fresh-water sources and safe, ‘toxin-free’ shellfish and fish stocks. To monitor these resources carefully, there is a need for the development of an in-situ algal-toxin monitoring solutions. Presented here is one of these solutions; The ToxiSense Detection system (Figure 1). This system has been designed to be as generically applicable to toxin detection as possible, with the cyanobacterial Microcystin-LR used for proof-of-concept.

Globally, the most prevalent cyanobacterial toxins, in blooms from fresh and brackish waters, are the cyclic peptide toxins of the microcystin family. The need for on-site algal-toxin monitoring has become increasingly urgent due to the amplified demand for fresh-water and for safe, ‘toxin-free’ shellfish and fish stocks. Along with routine testing of shellfish stocks for biotoxins including microcystin, the EU also require routine monitoring for the presence of the causative algal species. Herein, we describe a novel, Lab-On-A-Disc (LOAD) platform which has been developed to assess microcystin toxin levels in-situ. Using recombinant antibody technology, the LOAD platform combines immunofluorescence with centrifugally driven microfluidic liquid handling to achieve a next-generation disposable device for high throughput sampling. A low-complexity, ‘LED-Photodiode’ based optical sensing system was tailor-made for the system, which allows the fluorescence signal of the microcystin-specific reaction to be quantified. This system can rapidly and accurately detect the presence of microcystin in less than 10 minutes with minimum user interaction and maximum reproducibility. This method provides a low cost diagnostic alternative to the current laborious and costly methods used for toxin monitoring.

Episodes of toxin-producing phytoplankton occur worldwide, causing both animal and human fatalities. Toxicity occurs through consumption of phycotoxins, including azaspiracid, which accumulate in filter-feeding shellfish. Microcystins are hepatotoxins, produced mainly by freshwater cyanobacteria. Aflatoxins are potent, fungal hepatocarcinogens, which occur mainly in food and feed products. The purpose of this research was to examine the cytotoxic and immunosuppressive effects of aflatoxins (B1, B2 and G1), azaspiracid-1 and microcystin-LR in vitro, using the murine macrophage cell line, J774A.1. The results clearly demonstrated that azaspiracid and microcystin had a significant effect on host defence functions, through deregulation of IL-6, IL-10, IL12p40 and TNF-α cytokine expression. Microcystin exposure significantly decreased IL-1β expression. ‘Toll-like’ receptor (TLR2 and CD14) expression was altered following aflatoxin exposure, while apoptotic marker (caspase-1) expression was affected following microcystin exposure. This knowledge should be taken into consideration in the implementation of detection limits, aimed at minimising risks to human health through toxin exposure. Increased awareness of the hazards presented by toxins led to the requirement for recombinant antibodies for these targets, for incorporation into sensitive detection immunoassays. This thesis describes the production of leprine and avian immune libraries for azaspiracid and microcystin, respectively. Attempts were made to isolate azaspiracid-specific antibodies with little success. Phage display was utilised to successfully isolate two single chain antibody fragments (scFvs) to microcystin from the avian library. Error-prone PCR resulted in the isolation of a mutant clone which displayed a 2.3-fold improvement in sensitivity by ELISA, with an LOD of 1.4 ng/mL. The mutant scFv displayed an altered cross-reactivity profile to the microcystin variants tested using Biacore™ inhibition analysi

In this article we introduce a novel technology that utilizes specialized water dissolvable thin films for valving in centrifugal microfluidic systems. In previous work (William Meathrel and Cathy Moritz, IVD Technologies, 2007), dissolvable films (DFs) have been assembled in laminar flow devices to form efficient sacrificial valves where DFs simply open by direct contact with liquid. Here, we build on the original DF valving scheme to leverage sophisticated, merely rotationally actuated vapour barriers and flow control for enabling comprehensive assay integration with low-complexity instrumentation on "lab-on-a-disc" platforms. The advanced sacrificial valving function is achieved by creating an inverted gas-liquid stack upstream of the DF during priming of the system. At low rotational speeds, a pocket of trapped air prevents a surface-tension stabilized liquid plug from wetting the DF membrane. However, high-speed rotation disrupts the metastable gas/liquid interface to wet the DF and thus opens the valve. By judicious choice of the radial position and geometry of the valve, the burst frequency can be tuned over a wide range of rotational speeds nearly 10 times greater than those attained by common capillary burst valves based on hydrophobic constrictions. The broad range of reproducible burst frequencies of the DF valves bears the potential for full integration and automation of comprehensive, multi-step biochemical assay protocols. In this report we demonstrate DF valving, discuss the biocompatibility of using the films, and show a potential sequential valving system including the on-demand release of on-board stored liquid reagents, fast centrifugal sedimentation and vigorous mixing; thus providing a viable basis for use in lab-on-a-disc platforms for point-of-care diagnostics and other life science applications.