Learn how to measure protein aggregation and analyze subvisible particles, two of the key challenges in the development of biotherapeutics. Reasons for the selection of each of these particle types are explained. The horizon system can run 200 stability test samples from your formulation screen in less than a day, with just 25 l per test. Astm e3060 16 standard guide for subvisible particle. Halo labs subvisible particle analysis instrumentation. Part 2, experience with the application of subvisible particle analysis. In addition to the particlespecific analyses, a forensic case study was performed as an example of how a forensic lab may begin to incorporate subvisible particles into current schemes of analysis. The analysis of particulates has been a longstanding challenge in biopharmaceutical drug product development and quality control because the active constituents themselves may form particulate matter as a degradation product that may be difficult to quantify. Protein aggregation, proteinaceous particles, nonproteinaceous particles, subvisible particles, mfi, mvas, lumetics link, discriminant analysis. How the flowcam works for characterization of visible and subvisible particles turning data into insight. Overlooking subvisible particles in therapeutic protein. For the determination of particulate contamination 2procedures,method1lightobscuration particle count. Analytical methods to characterize aggregates and subvisible. Variable threshold method for determining the boundaries of imaged subvisible particles.
Subvisible particulate matter testing achieving reliable particle count and size analysis of injections, ophthalmic solutions, and lipid emulsions the united states pharmacopeia usp and other agencies such as ep, jp, and chp have established test methods to assure the minimization of particle counts in intravenous injections paren. The derived diameters and particle concentrations for the 20 lots of bcg it are summarized in table 5, and were consistent across the 20 lots analyzed. Before sample analysis, system suitability was confirmed for accurate sizing using a 1. With a strong focus on analysis of subvisible particles svp we feature the. Horizon subvisible particle analysis instrument halo labs. Analysis of the particle size distribution for all samples s1s5 and the reference bevacizumab revealed that, in general, the number of subvisible particles between 10 and 24. Highresolution images of subvisible particles between 1. New applications for the flowcam to be used for particle analysis in general industry are constantly developed. The green circle illustrates a large particle that the software clipped into many small particles.
Subvisible particles, visible particles, light obscuration, flow imaging, particle identification. Combining the highest specification technology with advanced software and automation options make these the most advanced instruments available for usp particle testing. New tools and opportunities presented by danny chou, president and founder at compassion biosolution, llc followed by an industry perspective presented by mark bumiller, technology manager at particle sizing systems, an entegris company. Visible and subvisible particles in the bcg immunotherapeutic. Analysis of subvisible particles pharmaceutical technology. Dynamic image analysis to evaluate subvisible particles. Characterization of visible and subvisible particles. The uncertainties around subvisible particles svps may warrant their being controlled as a critical quality attribute, fda office of biotechnology products division of therapeutic proteins dtp deputy director barry cherney stressed at a. Due to the morphology of the particles, modern application software makes it possible to determine particle characterization by number and size. Usp chapter, subvisible particulate matter in therapeutic protein injections, became official august 1, 2014 and provides specific guidance for proteinbased formulations. In this study adeno associated virus aav particles contaminated by host cell proteasomes are analysed with direct versus indirect methods performed in a standard. Mfi software gives you the ability to dive deep into a single sample or examine a.
Particle analysis can feel like finding a needle in a haystack, especially when youre dealing with subvisible particles. At gsk, imaging particle analysis using flowcam has been integrated into its testing program for development of proteinbased. The images collected are used by the software to create an extensive database of information size, shape. The flowcam captures highresolution, digital images of particles suspended in a fluid. Subvisible particle analysis in parenteral formulations. Archimedes is an innovative, highperformance system which harnesses the technique of resonant mass measurement in order to detect and count particles in the size range 60nm 5m and to measure particle mass and size. Measuring the size and concentration of subvisible particles within a. In case of ri influences, we recommend also using a lightindependent technique such as resonant mass measurement rmm archimedes for subvisible particle analysis in protein formulations. Protein aggregate and subvisible particles analysis.
Uncertainties in subvisible particulates need to be. Gustaf kylberg, rickard nordstrom and idamaria sintorn vironova ab, stockholm, sweden characterization and analysis of subvisible particles of biological origin can be challenging or give insufficient information. The horizon system can run 200 stability test samples from your formulation screen in less than a. A comparison of background membrane imaging versus flow. Evaluation of particle techniques for the characterization of.
For particle concentration, derived diameters d10 and d50, the coef. The laboratory instruments svss and sbss are now compatible to the pharmacopoeia usp 787 subvisible particulate matter in therapeutic protein injections. Subvisible particulate matter in therapeutic protein injections. The technical parameters and system settings of the. Uncertainties in subvisible particulates need to be understood, fda tells biotech cmc strategy forum. Typically, these particles are quantified by counting the number of particles in given size ranges. Oct 22, 2015 nanoparticle tracking analysis nta as the submicron fraction of the subvisible particle size range comes under greater scrutiny, techniques that enable the enumeration and sizing of nanoparticles are also increasingly vital additions to the characterisation toolkit. And youll have visual verification on the spot, as images are displayed in realtime. The case study involved the investigation of a subvisible paint particle.
Subvisible particle analysis shouldnt require milliliters of precious sample volume. Paswin particle analysis software the particle analyzing software paswin has been developed for particle measuring devices and analyzes particle size distributions of dispersed materials in liquids and gases and runs under windows. Mvss software is used to process the captured images and determine the particle size and count within a desired particle size range. Ranking your formulations based on a quantitative count of proteinaggregates shouldnt take weeks. The results obtained in examining a discrete unit or group of units for particulate matter cannot be extrapolated with certainty. Viavi micronir pro software provides data acquisition and chemometric model building, and assures compliance with title 21 cfr part 11, usp chapter 1119 and ep chapter 2.
Every particle in every image is then analyzed to create a database of particle count, size, transparency and morphology or shape. Subvisible and visible particle analysis in biopharmaceutical research and development this chapter gives an overview of methods for the analysis of subvisible and visible particles within. The guide will address the following components of particle analysis using dynamic imaging microscopy. Due to the complex milieu of subvisible particles, sample preparation and the method of analysis can affect the accuracy of. Expert knowhow in aggregate and subvisible particle analysis as a gmp compliant service. Dynamic image analysis to evaluate subvisible particles during continuous drug infusion in a neonatal intensive care unit. Nov 20, 2018 subscribe to cleanroom technology magazine for incisive analysis of the latest news and developments in hitech industries manufacturing in controlled environments. From the characterization of superabrasives, chemicals, petroleum, and fruit juice, to yeast and fragrance encapsulation if its a fluid, or can be suspended in a fluid, flowcam it. New tools and opportunities danny chou, president and founder at compassion biosolution, and mark bumiller. By contrast, the human eye can only count up to a limited number of particles figure 2. Protein aggregate and subvisible particles analysis information. The comfortable software supports several particle measuring devices. The horizon system is powered by a novel and highly sensitive backgrounded membrane imaging bmi technology.
Once a successful assay has been developed and validated, the resulting protocol, including run parameters, software. Quality of bevacizumab compounded for intravitreal administration. The accusizer system is used for usp,, and subvisible particulate matter testing. The bmi technology allows customers to image the particles on top of a coated membrane as if it were a conventional microscope slide. A gsk team member with expertise in particle analysis reasoned an imaging particle analyzer that takes digital images of each individual particle detected in a. Nanoparticle tracking analysis nta is one such technique.
Mvas software is also used to identify the sample particles morphology and shape attributes that traditional light obscuration lo and size exclusion. Particulate matter and particle size testing eurofins. Sub visible particle analysis shouldnt require milliliters of precious sample volume. These analytical challenges were met with success as long as the definition of particulate matter remained well within the. Subvisible particles are usually defined as particles that are too large for analysis by size exclusion chromatography sec e. Subvisible particles particulate contamination of injections and infusions consists of extraneous, mobile undissolved particles, other than gas bubbles, unintentionally present in the solutions.
Why simply counting shadows leaves you in the dark about us the downstream column is a blog and community for scientists and others involved in downstream biomanufacturing. An accurate assessment of particle characteristics and concentrations in pharmaceutical products by flow imaging requires accurate particle sizing and morphological analysis. Separation, characterization and discriminant analysis of. Usp, therapeutic protein injections beckman coulter. Excessive particulate contamination in proteinbased drug products may lead to adverse health outcomes. Powders for parenteral use are reconstituted with particle free water or with an. Mfi has become an established technique for subvisible particle analysis, though no monographspecifications in the pharmacopoeias exist so far. Subvisible particle analysis the potential immunogenicity of particles ranging from nanometer to micron sizes in biotherapeutics has been a growing area of discussion between regulators and industry. Transfer to the filtration funnel the total volume of a solution pool or. Coriolis has received cgmp accreditation by the local regulatory authority and since then we have been reinspected by the authority and audited by several clients successfully. Critical gaps in the analysis and control of subvisible particles. Application software permits the precise evaluation of visible mobile and immobile particles in each unit. The flow imaging method of particle size analysis relies on the definition of the particle. Particle morphology, captured by images, can be used to determine whether the svp is inherent, intrinsic, or extrinsic.
The accusizer tm line of instruments are based on the principle of single particle optical sizing spos. To achieve unparalleled subvisible particle analysis, the horizon system employs halo labs proprietary analytical technique backgrounded membrane imaging bmi. This chapter gives an overview of methods for the analysis of subvisible and visible particles within biopharmaceutical products. Images of the sample are captured as it passes through the flow cells sensing zone. Archimedes is uniquely positioned to quantitatively measure protein aggregates in biotherapeutics in the size range 200nm. Each particle image is saved to a userbuilt reference library and used to improve the software s pattern recognition algorithms and improve particle identification. Monoclonal antibodies mab and related products are widely used as therapeutic agents to treat various cancers and autoimmune diseases.
The field of particle analysis is highly dynamic and various emerging techniques are available for this purpose. The instructional chapter usp mentions mfi as a possible orthogonal technique for subvisible. Subvisible particle analysis protein antibody integrity bio. Because its imagebased, you can add quantifiable morphological parameters to your analysis. The presence of subvisible particles svps in parenteral formulations of biologics is a major challenge in the development of therapeutic protein formulations. Subvisible and visible particle analysis in biopharmaceutical. Particle count test and method 2 microscopic particle count test, are specified hereinafter.
Microflow imaging imagebased particle analysis for biopharmaceuticals. The calibration records are retained by the software and can be retrieved for subsequent uses. The production of mab therapeutics can be highly challenging due to various chemical degradation processes and aggregation. In this study adeno associated virus aav particles contaminated by host cell proteasomes are analysed with direct versus indirect methods continued.
Particulate testing solutions for contamination control. How does dynamic imaging particle analysis allow identification of subvisible particles. Within this section, a brief overview of emerging or less routinely used techniques for subvisible particle analysis is provided. The instructional chapter usp mentions mfi as a possible orthogonal technique for subvisible particle analysis in addition to light obscuration. Analysis of images begins with the definition of particle boundaries from the captured. Usp related products and solutions beckman coulter. Entegris manufactures a range of particle characterization instrumentation for both the laboratory and online monitoring.
Smallvolume parenterals having a volume of 25 ml or more may be tested individually. By enabling low volume and high throughput subvisible particle analysis, the horizon system makes it possible to perform particle screening as early as. With walkaway automation, the horizon instrument helps you screen up to 96 formulation samples in under 2 hours with far superior particle image analysis than current methods. Archimedes subvisible particle detection, counting.
A comparison of background membrane imaging versus flow technologies for subvisible particle analysis of biologics. Achieving reliable particle count and size analysis of. Archimedes, an innovative system for detection and counting of subvisible particles including protein aggregates and for particle mass and size measurement. Astm e306016, standard guide for subvisible particle measurement in biopharmaceutical manufacturing using dynamic flow imaging microscopy, astm international, west conshohocken, pa, 2016, back to top. The subvisible and visible particles present in a solution are often classified based on size, and are quantified by the actual number of particles present rather than by weight or molar amounts. Particle counting and analysis techniques are used to measure particle contamination and ensure adherence to specified limits. The software can also be used to validate particle counters. Variable threshold method for determining the boundaries of imaged subvisible particles r. Particle size was calculated based on equivalent circular diameter using the mfi software. Overlooking subvisible particles in therapeutic protein products. Collaborative study for analysis of subvisible particles using flow. The horizon instrument is the industrys first system designed specifically to address the demand for quantitative subvisible particle analysis at every stage of biologic drug formulation.
In this study adeno associated virus aav particles contaminated by host cell proteasomes are analysed with direct versus indirect methods performed in. Quality of bevacizumab compounded for intravitreal. The ability to see the actual particles in parenteral formulations, to see their morphology, and to automatically differentiate one type from another marks an important advance in parental drug particle analysis. So the user can see at one glance whether the analysed sample meets the requirements of the selected standard.
Postprocessing analysis provides the ability to create image libraries, identify and exclude certain types of particulates silicone oil for one example and create more accurate particle counts. Light obscuration is the method of choice listed in the pharmacopoeias for the analysis of subvisible particles in parenteral products. Analysis of images begins with the definition of particle boundaries. Imagebased particle analysis for biopharmaceuticals. Commonly a single threshold defines the level for a pixel in the image. The digital images of the particles present in the sample are processed by image morphology analysis software that allows their quantification in size and count. Mfi gives you more insight into the world of subvisible particlesway more than just size and count.
Subscribe to cleanroom technology magazine for incisive analysis of the latest news and developments in hitech industries manufacturing in controlled environments. Protein aggregate and subvisible particles analysis information thermo fisher scientific us. Aug 24, 2017 dynamic image analysis to evaluate subvisible particles during continuous drug infusion in a neonatal intensive care unit. When examining injections and parenteral infusions for subvisible particles. Direct imaging particle measurement technologies have. Variable threshold method for determining the boundaries. Mfi software provides you with the tools to get the job done. Variable threshold method for determining the boundaries of. Analysis of subvisible particles and aggregates in gmp.
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