News & Views
Interview Stabilization of biological assays: conventional versus new technology
Miles Burrows speaks to Lauren Constable, Head of Commissioning Following completion of his PhD in Chemistry at Cardiff University (UK), Miles Burrows started his career at Amersham Biosciences (UK), predominately researching fluorescent reagents and the labeling of biological compounds. Following the acquisition of Amersham by GE Healthcare (UK), Burrows took on the role of R&D Technology Manager, leading a multifunctional team of chemists, biologists and industrial design engineers through new product introduction, research and product care projects. During this period, Burrows oversaw the launch of a custom assay stabilization service based on the well-established technology as used in GE Healthcare’s Ready-To-Go™ product range. Since 2012 Burrows has been the Global Product Manager for custom molecular biology and fluorescent reagents for GE Healthcare Life Sciences. In what context related to biomarker analysis & molecular diagnostics is the stabilization of biological assays used?
We see increasing use in stabilization of biological assays in two main areas. First, within point-of-care or near-use assays following the increasing shift from the centralized laboratory model to a field-based use model by molecular diagnostics assay providers. There are two logistical headaches that need to be overcome when developing a near application use assay. The first is the maintenance of the cold chain or elimination of refrigerated storage and the second is the requirement for a simplified workflow. In addition, within a laboratory setting, we are also seeing the increased use of automated workflows, ‘deskilling’ of the laboratory and increased use of high-throughput assays. Therefore, the ability to store online, on platforms outside of refrigerated storage and with simplified liquid handling requirements is an area of increasing importance where the stabilized and simplified assays are also being used. As a general trend, we see increased demand for both reagent and sample stabilization technologies, driven by companies striving to increase access to molecular diagnostic tests while in parallel lowering the costs of their assays.
GE Healthcare UK Limited, Amersham Place, Little Chalfont, Buckinghamshire HP7 9NA, UK [email protected]
Lyophilization is one of the key steps within our Ready-To-Go™ (RTG™) product. It is a well-understood and robust technique for stabilization of biologicals. However, simply freeze-drying a wet mix will not yield a stabilized assay. In particular, the product from a classical lyophilization approach does not necessarily yield the final format that is required by developers of molecular assays for near-application use. For example, there is a requirement to balance fast dissolution rates with a robust product format that can be manipulated by hand or by instrumentation if necessary. It is also key to use lyoprotectants and excipients that do not interfere with either the assay performance or signal reads. The fine powders that are more traditionally associated with freeze-drying are less amenable to incorporation into an automated workflow for use outside of a laboratory setting. What is RTG ambient temperature stabilization technology?
the limitations of conventional lyophilization techniques? How do these impact manufacturers & users?
RTG is a technology that has been designed to overcome the two main barriers associated with use of molecular diagnostics in field and near-application settings. First, by formulating a one-shot assay, it simplifies the preparation steps, which allows for a less convoluted workflow in the field and reduces the risk of quality issues being introduced by the operator. Second, the ambient temperature stability removes the requirement for the cold chain and increases the potential geographical reach of the technology.
10.2217/BMM.13.96 © 2013 Future Medicine Ltd
Biomarkers Med. (2013) 7(6), 929–931
What are widely considered to be
Miles Burrows
part of
ISSN 1752-0363
929
News & Views – Interview Specifically, the product is a complete assay wet mix, formulated with our proprietary lyoprotectants and excipients to yield the final format, which is an amorphous glass in nature. As an amorphous glass, the product characteristics can be controlled to yield optimal properties for molecular diagnostic assays, in particular a key parameter of the product is the glass transition temperature – this is essentially the temperature at which the glass starts to melt and the product returns to a liquid state. Typically, our assays produce transition temperatures of at least 55°C or greater, which means that, provided the product does not exceed 55°C, the integrity of the product is maintained. Geographical reach is clearly one issue associated with cold chain maintenance & shipping on dry ice: what challenges are currently faced with shipping assays on dry ice & how does this impact both manufacturers & end users?
There are the logistical challenges associated with re-icing packages – especially in more remote geographical locations. Getting shipments across borders and through customs without delays is not always straightforward. Ensuring that these often precious biological materials are kept intact without thawing if an unplanned delay has occurred is difficult. A stabilized format ensures the integrity of product through the supply chain; the questions around product viability during shipping simply do not arise anymore. Then there is the cost associated with shipping on dry ice. The packages are bulkier, heavier and require special hand ling, all of which add cost. An internal GE study revealed that the cost associated with shipping 14,000 items of various sizes on dry ice to global destinations from our Uppsala (Sweden) warehouse was approximately US$1.4 million. This included costs for packing materials (US$400,000), labor (US$120,000), transport to Europe (US$150,000) and the rest of the world (US$300,000) from Sweden, and an estimate of the USA and Canada plus the Asia Pacific (US$400,000). Furthermore, some of our customers have told us, anecdotally, that a third of their assay production costs are associated with shipping on dry ice. 930
Biomarkers Med. (2013) 7(6)
Other than shipping, what do you consider to be the benefits of RTG over conventional techniques?
Our RTG products balance essential characteristics of a stabilized assay product format. They have a long ambient temperature shelf life (e.g., we have three year ambient temperature stability data for our polymerase enzyme), and a product format that is robust enough to remain intact and in place during transportation. A stable and transportable format is critical as more and more assays are housed within customized plasticware that requires the product to be delivered at specific positions within instrumentation – this is achievable with the RTG format by manufacturing the assay directly into the housing. In addition, rapid reconstitution rates of the RTG reagent format are achieved without the requirement to mix or to vortex. Further more, we have proven that our RTG lyoprotectants and excipients are compatible with PCR assay formats (Polymerase Chain Reaction [PCR] is covered by patents owned by Roche Molecular Systems and F Hoffman-La Roche Ltd.) and do not interfere with the assay chemistry. RTG seems to be an extra step in the workflow, can this be justified?
It is an extra step in development, but not an extra step in the user’s workflow, and I think that is a critical distinction to make. In fact, the format is a considerable reduction in the workflow to those performing the assay. By investing upfront to develop the correct format, developers can access wider markets, but more importantly, stabilized formats will become the industry norm. We are already seeing established assay providers recognizing the benefits of switching from a frozen or liquid format and I think that there is danger of those who do not make the switch getting left behind by an industry that is advancing and differentiating. Looking to the future, how can you see RTG impacting the current field?
In general, any process that has requirements for a cold chain or applications with a requirement to reduce the complexity of the assay could benefit from RTG. The technology is currently being applied to future science group
Interview – the fields of pathogen detection, virus identification, food testing, veterinary applications, and security or biothreat applications. The security and biothreat applications are a good example of where a simplified workflow is required, as users typically have limited mobility owing to the protective equipment worn. But, I think that the biggest impact is being made where the stabilized reagents are aligned with developments in instrument technology. We are now seeing that there are truly portable nucleic acid testing platforms capable of delivering results within the time frame of a couple of hours. These instruments will change the way diagnostic tests are conducted and increase access of the global population to testing. In order to maximize the benefits afforded by the developments in instrumentation, the assay format has to meet the challenges of
future science group
News & Views
simplification and stabilization, which is where RTG comes in. Disclaimer The opinions expressed in this interview are those of the interviewee and do not necessarily reflect the views of Future Medicine Ltd.
Financial & competing interests disclosure M Burrows has been an employee of the General Electric Company via acquisition of Amersham Biosciences since 2001. Ready-To-Go and RTG are trademarks of GE Healthcare companies. The author has no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript.
www.futuremedicine.com
931
Interview Stabilization of biological assays: conventional versus new technology
Miles Burrows speaks to Lauren Constable, Head of Commissioning Following completion of his PhD in Chemistry at Cardiff University (UK), Miles Burrows started his career at Amersham Biosciences (UK), predominately researching fluorescent reagents and the labeling of biological compounds. Following the acquisition of Amersham by GE Healthcare (UK), Burrows took on the role of R&D Technology Manager, leading a multifunctional team of chemists, biologists and industrial design engineers through new product introduction, research and product care projects. During this period, Burrows oversaw the launch of a custom assay stabilization service based on the well-established technology as used in GE Healthcare’s Ready-To-Go™ product range. Since 2012 Burrows has been the Global Product Manager for custom molecular biology and fluorescent reagents for GE Healthcare Life Sciences. In what context related to biomarker analysis & molecular diagnostics is the stabilization of biological assays used?
We see increasing use in stabilization of biological assays in two main areas. First, within point-of-care or near-use assays following the increasing shift from the centralized laboratory model to a field-based use model by molecular diagnostics assay providers. There are two logistical headaches that need to be overcome when developing a near application use assay. The first is the maintenance of the cold chain or elimination of refrigerated storage and the second is the requirement for a simplified workflow. In addition, within a laboratory setting, we are also seeing the increased use of automated workflows, ‘deskilling’ of the laboratory and increased use of high-throughput assays. Therefore, the ability to store online, on platforms outside of refrigerated storage and with simplified liquid handling requirements is an area of increasing importance where the stabilized and simplified assays are also being used. As a general trend, we see increased demand for both reagent and sample stabilization technologies, driven by companies striving to increase access to molecular diagnostic tests while in parallel lowering the costs of their assays.
GE Healthcare UK Limited, Amersham Place, Little Chalfont, Buckinghamshire HP7 9NA, UK [email protected]
Lyophilization is one of the key steps within our Ready-To-Go™ (RTG™) product. It is a well-understood and robust technique for stabilization of biologicals. However, simply freeze-drying a wet mix will not yield a stabilized assay. In particular, the product from a classical lyophilization approach does not necessarily yield the final format that is required by developers of molecular assays for near-application use. For example, there is a requirement to balance fast dissolution rates with a robust product format that can be manipulated by hand or by instrumentation if necessary. It is also key to use lyoprotectants and excipients that do not interfere with either the assay performance or signal reads. The fine powders that are more traditionally associated with freeze-drying are less amenable to incorporation into an automated workflow for use outside of a laboratory setting. What is RTG ambient temperature stabilization technology?
the limitations of conventional lyophilization techniques? How do these impact manufacturers & users?
RTG is a technology that has been designed to overcome the two main barriers associated with use of molecular diagnostics in field and near-application settings. First, by formulating a one-shot assay, it simplifies the preparation steps, which allows for a less convoluted workflow in the field and reduces the risk of quality issues being introduced by the operator. Second, the ambient temperature stability removes the requirement for the cold chain and increases the potential geographical reach of the technology.
10.2217/BMM.13.96 © 2013 Future Medicine Ltd
Biomarkers Med. (2013) 7(6), 929–931
What are widely considered to be
Miles Burrows
part of
ISSN 1752-0363
929
News & Views – Interview Specifically, the product is a complete assay wet mix, formulated with our proprietary lyoprotectants and excipients to yield the final format, which is an amorphous glass in nature. As an amorphous glass, the product characteristics can be controlled to yield optimal properties for molecular diagnostic assays, in particular a key parameter of the product is the glass transition temperature – this is essentially the temperature at which the glass starts to melt and the product returns to a liquid state. Typically, our assays produce transition temperatures of at least 55°C or greater, which means that, provided the product does not exceed 55°C, the integrity of the product is maintained. Geographical reach is clearly one issue associated with cold chain maintenance & shipping on dry ice: what challenges are currently faced with shipping assays on dry ice & how does this impact both manufacturers & end users?
There are the logistical challenges associated with re-icing packages – especially in more remote geographical locations. Getting shipments across borders and through customs without delays is not always straightforward. Ensuring that these often precious biological materials are kept intact without thawing if an unplanned delay has occurred is difficult. A stabilized format ensures the integrity of product through the supply chain; the questions around product viability during shipping simply do not arise anymore. Then there is the cost associated with shipping on dry ice. The packages are bulkier, heavier and require special hand ling, all of which add cost. An internal GE study revealed that the cost associated with shipping 14,000 items of various sizes on dry ice to global destinations from our Uppsala (Sweden) warehouse was approximately US$1.4 million. This included costs for packing materials (US$400,000), labor (US$120,000), transport to Europe (US$150,000) and the rest of the world (US$300,000) from Sweden, and an estimate of the USA and Canada plus the Asia Pacific (US$400,000). Furthermore, some of our customers have told us, anecdotally, that a third of their assay production costs are associated with shipping on dry ice. 930
Biomarkers Med. (2013) 7(6)
Other than shipping, what do you consider to be the benefits of RTG over conventional techniques?
Our RTG products balance essential characteristics of a stabilized assay product format. They have a long ambient temperature shelf life (e.g., we have three year ambient temperature stability data for our polymerase enzyme), and a product format that is robust enough to remain intact and in place during transportation. A stable and transportable format is critical as more and more assays are housed within customized plasticware that requires the product to be delivered at specific positions within instrumentation – this is achievable with the RTG format by manufacturing the assay directly into the housing. In addition, rapid reconstitution rates of the RTG reagent format are achieved without the requirement to mix or to vortex. Further more, we have proven that our RTG lyoprotectants and excipients are compatible with PCR assay formats (Polymerase Chain Reaction [PCR] is covered by patents owned by Roche Molecular Systems and F Hoffman-La Roche Ltd.) and do not interfere with the assay chemistry. RTG seems to be an extra step in the workflow, can this be justified?
It is an extra step in development, but not an extra step in the user’s workflow, and I think that is a critical distinction to make. In fact, the format is a considerable reduction in the workflow to those performing the assay. By investing upfront to develop the correct format, developers can access wider markets, but more importantly, stabilized formats will become the industry norm. We are already seeing established assay providers recognizing the benefits of switching from a frozen or liquid format and I think that there is danger of those who do not make the switch getting left behind by an industry that is advancing and differentiating. Looking to the future, how can you see RTG impacting the current field?
In general, any process that has requirements for a cold chain or applications with a requirement to reduce the complexity of the assay could benefit from RTG. The technology is currently being applied to future science group
Interview – the fields of pathogen detection, virus identification, food testing, veterinary applications, and security or biothreat applications. The security and biothreat applications are a good example of where a simplified workflow is required, as users typically have limited mobility owing to the protective equipment worn. But, I think that the biggest impact is being made where the stabilized reagents are aligned with developments in instrument technology. We are now seeing that there are truly portable nucleic acid testing platforms capable of delivering results within the time frame of a couple of hours. These instruments will change the way diagnostic tests are conducted and increase access of the global population to testing. In order to maximize the benefits afforded by the developments in instrumentation, the assay format has to meet the challenges of
future science group
News & Views
simplification and stabilization, which is where RTG comes in. Disclaimer The opinions expressed in this interview are those of the interviewee and do not necessarily reflect the views of Future Medicine Ltd.
Financial & competing interests disclosure M Burrows has been an employee of the General Electric Company via acquisition of Amersham Biosciences since 2001. Ready-To-Go and RTG are trademarks of GE Healthcare companies. The author has no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript.
www.futuremedicine.com
931
