Analytical Biotechnology

Rye, MD, United States

Analytical Biotechnology

Rye, MD, United States
SEARCH FILTERS
Time filter
Source Type

Mathonet S.,CMC Biologics | Mahler H.-C.,Lonza AG | Esswein S.T.,Abbvie Deutschland GmbH and Co. KG | Mazaheri M.,Analytical Biotechnology | And 6 more authors.
PDA Journal of Pharmaceutical Science and Technology | Year: 2016

Regulatory monographs in Europe and the United States require drug products for parenteral administration to be "practically free" or "essentially free" of visible particles, respectively. Both terms have been used interchangeably and acknowledge the probabilistic nature of visual particle inspection. The probability of seeing a particle in a drug product container varies according to the size and nature of the particles as well as container and inspection conditions. Therefore, the term "without visible particles" can be highly misleading in the context of what is practically achievable. This may lead to differences in understanding between industry practitioners and regulatory agencies. Is this term intended to mean "zero particles", or is there any intention to distinguish between particle type such as "zero extraneous visible particles" or "zero proteinaceous particles"? Furthermore, how can "zero" particles as a criterion for release testing be reconciled with "practically free from particles" as stated in the definition and a low, justified level of proteinaceous particles after production? The purpose of this position paper is to review best practices in the industry in terms of visual inspection process and associated operator training, quality control sampling, testing, and setting acceptance criteria corresponding to "practically free of visible particles" and providing considerations when visible proteinaceous particles are deemed unavoidable. It also provides a brief overview of visible particle characterization and gives perspectives on patient safety. This position paper applies to biotechnology-derived drug products including monoclonal antibodies in late-phase development to licensed products.


Bee J.S.,Formulation science | Frey V.V.,Analytical Biotechnology | Javed U.,Analytical Biotechnology | Chung J.,Drug Delivery and Devices | And 6 more authors.
PDA Journal of Pharmaceutical Science and Technology | Year: 2014

Glass prefillable syringes are lubricated with silicone oil to ensure functionality and a consistent injection for the end user. If excessive silicone is applied, droplets could potentially result in aggregation of sensitive biopharmaceuticals or clouding of the solution. Therefore, monitoring and optimization of the applied silicone layer is critical for prefilled syringe development. The hydrophobic properties of silicone oil, the potential for assay interference, and the very small quantities applied to prefilled syringes present a challenge for the development of a suitable assay. In this work we present a rapid and simple Fourier transform infrared (FTIR) spectroscopy method for quantitation of total silicone levels applied to prefilled syringes. Leveldependent silicone oil migration occurred over time for empty prefilled syringes stored tip-up. However, migration from all prefilled syringes with between 0.25 and 0.8 mg of initial silicone oil resulted in a stable limiting minimum level of between 0.15 and 0.26 mg of silicone in the syringe reached after 1 to 4 years of empty tip-up storage. The results of the FTIR assay correlated well with non-destructive reflectometry characterization of the syringes. This assay can provide valuable data for selection of a robust initial silicone oil target and quality control of prefilled syringes intended for biopharmaceuticals. ©PDA, Inc. 2014.


Cash P.W.,Analytical Biotechnology | Narwal R.,Formulation science | Levitskaya S.V.,Analytical Biotechnology | Krause S.,Analytical Biotechnology | Mazaheri M.,Analytical Biotechnology
PDA Journal of Pharmaceutical Science and Technology | Year: 2016

Visible particles must be monitored as part of the control strategy for pharmaceutical products. Extraneous (foreign) particles are not acceptable in parenteral drug products. In biopharmaceuticals, formation of protein particles is recognized as an inherent quality attribute. All protein therapeutics contain particles that vary greatly in visibility and size from invisible (sub-micron) to visible (millimeter) and, as part of the control strategy, biopharmaceutical companies are required to monitor and minimize the presence of visible and sub-visible particles in their products. There is an industry-wide unmet need for particle standards for visual inspection of protein therapeutics. A new, semi-quantitative method using particle standards for assessing the levels of small, inherent visible particles is presented. This method can be used during product development to identify a formulation that minimizes particle formation and also during release and stability testing to monitor and control inherent proteinaceous visible particles. © PDA, Inc. 2016.


Liu P.M.,Global Bioassays and Technology | Zou L.,Global Bioassays and Technology | Sadhu C.,Analytical Biotechnology | Shen W.D.,NGM Biopharmaceuticals | Nock S.,Global Biologics
Bioanalysis | Year: 2015

An appropriate assessment strategy with validated anti-drug antibody (ADA) assays is critical for comparative evaluation of immunogenicity between a proposed biosimilar and its reference product. The strategy should aim to identify potential differences in immune responses between these products. While an ADA assay employing the proposed biosimilar product as the detecting reagent has been generally recommended for such evaluation, a product-specific assay using the product of interest may be of use as it offers a capability of detecting antibodies against specific epitopes from the respective product. Regardless of assay strategy, the performance of the assay must be fully assessed and method needs to be validated to meet the comparative purpose of immunogenicity assessment. © 2015 Future Science Ltd.


PubMed | Lonza AG, Amgen, Bristol Myers Squibb, Hoffmann-La Roche and 4 more.
Type: Journal Article | Journal: PDA journal of pharmaceutical science and technology | Year: 2016

Regulatory monographs in Europe and the United States require drug products for parenteral administration to be practically free or essentially free of visible particles, respectively. Both terms have been used interchangeably and acknowledge the probabilistic nature of visual particle inspection. The probability of seeing a particle in a drug product container varies according to the size and nature of the particles as well as container and inspection conditions. Therefore, the term without visible particles can be highly misleading in the context of what is practically achievable. This may lead to differences in understanding between industry practitioners and regulatory agencies. Is this term intended to mean zero particles, or is there any intention to distinguish between particle type such as zero extraneous visible particles or zero proteinaceous particles? Furthermore, how can zero particles as a criterion for release testing be reconciled with practically free from particles as stated in the definition and a low, justified level of proteinaceous particles after production?The purpose of this position paper is to review best practices in the industry in terms of visual inspection process and associated operator training, quality control sampling, testing, and setting acceptance criteria corresponding to practically free of visible particles and providing considerations when visible proteinaceous particles are deemed unavoidable. It also provides a brief overview of visible particle characterization and gives perspectives on patient safety. This position paper applies to biotechnology-derived drug products including monoclonal antibodies in late-phase development to licensed products.In the 2011 monoclonal antibody monograph revision, European Pharmacopoeia experts acknowledged that protein products may also contain proteinaceous particles at release or that protein particles may form during storage. Indeed, industry experience has demonstrated that therapeutic proteins such as monoclonal antibodies can exhibit a propensity for self-association leading to the formation of aggregates that range in size from nanometres (oligomers) to microns (subvisible and visible particles). As a result, the requirement for drug product appearance for monoclonal antibodies was changed from without visible particles to without visible particles unless otherwise authorised or justified. In our view, practically free from particles should be considered a suitable acceptance criterion for injectable biotechnology and small-molecule products, as long as appropriately defined. Furthermore, we argue that visual inspection is a suitable quality control release test and that practically free from particles is a suitable specification when adequately described.


PubMed | Formulation science, Analytical Biotechnology, Biogen Idec and Drug Delivery & Devices
Type: Journal Article | Journal: PDA journal of pharmaceutical science and technology | Year: 2014

Glass prefillable syringes are lubricated with silicone oil to ensure functionality and a consistent injection for the end user. If excessive silicone is applied, droplets could potentially result in aggregation of sensitive biopharmaceuticals or clouding of the solution. Therefore, monitoring and optimization of the applied silicone layer is critical for prefilled syringe development. The hydrophobic properties of silicone oil, the potential for assay interference, and the very small quantities applied to prefilled syringes present a challenge for the development of a suitable assay. In this work we present a rapid and simple Fourier transform infrared (FTIR) spectroscopy method for quantitation of total silicone levels applied to prefilled syringes. Level-dependent silicone oil migration occurred over time for empty prefilled syringes stored tip-up. However, migration from all prefilled syringes with between 0.25 and 0.8 mg of initial silicone oil resulted in a stable limiting minimum level of between 0.15 and 0.26 mg of silicone in the syringe reached after 1 to 4 years of empty tip-up storage. The results of the FTIR assay correlated well with non-destructive reflectometry characterization of the syringes. This assay can provide valuable data for selection of a robust initial silicone oil target and quality control of prefilled syringes intended for biopharmaceuticals.Glass prefillable syringes are lubricated with silicone oil to ensure functionality and a consistent injection for the end user. If excessive silicone is applied, droplets could potentially result in aggregation of sensitive biopharmaceuticals or clouding of the solution. Therefore, monitoring and optimization of the applied silicone layer is critical for prefilled syringe development. The hydrophobic properties of silicone oil, the potential for assay interference, and the very small quantities applied to prefilled syringes present a challenge for the development of a suitable assay. In this work we present a rapid and simple Fourier transform infrared (FTIR) spectroscopy method for quantitation of total silicone levels applied to prefilled syringes. Level-dependent silicone oil migration occurred over time for empty prefilled syringes stored tip-up. However, migration from all prefilled syringes with between 0.25 and 0.8 mg of initial silicone oil resulted in a stable limiting minimum level of between 0.15 and 0.26 mg of silicone in the syringe reached after 1 to 4 years of empty tip-up storage. The results of the FTIR assay correlated well with non-destructive reflectometry characterization of the syringes. This assay can provide valuable data for selection of a robust initial silicone oil target and quality control of prefilled syringes intended for biopharmaceuticals.


PubMed | Formulation science and Analytical Biotechnology
Type: Journal Article | Journal: PDA journal of pharmaceutical science and technology | Year: 2016

Visible particles must be monitored as part of the control strategy for pharmaceutical products. Extraneous (foreign) particles are not acceptable in parenteral drug products. In biopharmaceuticals, formation of protein particles is recognized as an inherent quality attribute. All protein therapeutics contain particles that vary greatly in visibility and size from invisible (sub-micron) to visible (millimeter) and, as part of the control strategy, biopharmaceutical companies are required to monitor and minimize the presence of visible and sub-visible particles in their products. There is an industry-wide unmet need for particle standards for visual inspection of protein therapeutics. A new, semi-quantitative method using particle standards for assessing the levels of small, inherent visible particles is presented. This method can be used during product development to identify a formulation that minimizes particle formation and also during release and stability testing to monitor and control inherent proteinaceous visible particles.Visible particles must be monitored as part of the control strategy for parenteral biopharmaceutical drug products. In these products, formation of protein particles is a natural occurrence. All protein drugs contain particles that vary greatly in visibility and size from invisible (sub-micron) to visible (millimeter), and pharmaceutical companies are required to monitor and minimize the presence of visible and sub-visible particles in their products. There is an industry-wide unmet need for particle standards for visual inspection of protein drugs. A new, semi-quantitative method using particle standards for assessing the levels of small, naturally occurring visible particles is presented. This method can be used during drug development to identify a formulation that minimizes particle formation and also during testing of final clinical or commercial drug product to monitor and control naturally occurring proteinaceous visible particles.


Liu H.,Analytical Biotechnology | Lei Q.P.,Analytical Biotechnology | Washabaugh M.,Analytical Biotechnology
Analytical Chemistry | Year: 2016

The complication of IgG2 disulfide connections demands advances in techniques for disulfide bond determination. We have developed a new LC/MS/MS method for improved disulfide analysis. With postcolumn introduction of dithiothreitol (DTT) and ammonium hydroxide, each disulfide-containing peptide eluted out of LC in an acidic mobile phase can be rapidly reduced prior to MS analysis. The reduction can be driven to near completion. The reagents are MS-friendly, and the reaction occurs at no cost of separation (little is added to the postcolumn dead volume of the LC system). Comparing LC/MS data with and without online reduction, a direct correlation can be established between a disulfide peptide and its composing peptides using retention time. With disulfide online removal, high-quality MS/MS fragmentation data can be acquired and allows for definitive determination of the disulfide peptide. This technique is especially valuable in determining the disulfide bond linkage of complicated molecules such as the hinge-containing disulfide peptides produced from IgG2 disulfide isoforms. Due to over/under enzymatic cleavages, multiple hinge-containing disulfide peptides are produced from each isoform. Twenty-two hinge-containing disulfide peptides in total have been confidently identified with this technique. Without the method, successful identification to many of these peptides would have become extremely difficult. © 2016 American Chemical Society.


PubMed | Analytical Biotechnology and Purification Process science
Type: Journal Article | Journal: Biotechnology progress | Year: 2015

Chinese hamster ovary (CHO) cells are often used to produce therapeutic monoclonal antibodies (mAbs). CHO cells express many host cell proteins (HCPs) required for their growth. Interactions of HCPs with mAbs can sometimes result in co-purification of trace levels of hitchhiker HCPs during the manufacturing process. Purified mAb-1 product produced in early stages of process optimization had high HCP levels. In addition, these lots formed delayed-onset particles containing mAb-1 and its heavy chain C-terminal fragments. Studies were performed to determine the cause of the observed particle formation and to optimize the purification for improved HCP clearance. Protease activity and inhibitor stability studies confirmed that an aspartyl protease was responsible for fragmentation of mAb-1 resulting in particle formation. An affinity resin was used to selectively capture aspartyl proteases from the mAb-1 product. Mass spectrometry identified the captured aspartyl protease as CHO cathepsin D. A wash step at high pH with salt and caprylate was implemented during the protein A affinity step to disrupt the HCP-mAb interactions and improve HCP clearance. The product at the end of purification using the optimized process had very low HCP levels, did not contain detectable protease activity, and did not form particles. Spiking of CHO cathepsin D back into mAb-1 product from the optimized process confirmed that it was the cause of the particle formation. This work demonstrated that process optimization focused on removal of HCPs was successful in eliminating particle formation in the final mAb-1 product.


PubMed | Analytical Biotechnology
Type: Journal Article | Journal: Analytical chemistry | Year: 2016

The complication of IgG2 disulfide connections demands advances in techniques for disulfide bond determination. We have developed a new LC/MS/MS method for improved disulfide analysis. With postcolumn introduction of dithiothreitol (DTT) and ammonium hydroxide, each disulfide-containing peptide eluted out of LC in an acidic mobile phase can be rapidly reduced prior to MS analysis. The reduction can be driven to near completion. The reagents are MS-friendly, and the reaction occurs at no cost of separation (little is added to the postcolumn dead volume of the LC system). Comparing LC/MS data with and without online reduction, a direct correlation can be established between a disulfide peptide and its composing peptides using retention time. With disulfide online removal, high-quality MS/MS fragmentation data can be acquired and allows for definitive determination of the disulfide peptide. This technique is especially valuable in determining the disulfide bond linkage of complicated molecules such as the hinge-containing disulfide peptides produced from IgG2 disulfide isoforms. Due to over/under enzymatic cleavages, multiple hinge-containing disulfide peptides are produced from each isoform. Twenty-two hinge-containing disulfide peptides in total have been confidently identified with this technique. Without the method, successful identification to many of these peptides would have become extremely difficult.

Loading Analytical Biotechnology collaborators
Loading Analytical Biotechnology collaborators