Antibody aggregation

In antibody aggregation studies, common methods include difference spectra, principal component analysis (PCA), partial least squares (PLS) and multivariate curve resolution (MCR) [22,23,40]. The application of chemometrics to Raman data is a broad area of research.

Mar 25, 2018 · The aggregation propensity of an antibody can be determined by its intrinsic properties such as primary sequence and structure. 12 Protein folds into a functional conformation that represents the lowest free energy and thus most stable, and this conformation is referred to as a native conformation. 13 Antibody at its native state has less ... This paper investigates how mAb aggregates generated by a variety of mechanical, thermal and chemical stresses impact the biological activity of a biotherapeutic. Increased aggregation resulted in a decrease in biological activity, as confirmed by cell based assays such as antibody dependent cell mediated cytotoxicity (ADCC) and complement ...However, the formulation with 100 mM Arg·Glu was more stable than the reference formulation, with Arg·Glu reducing the rate of antibody aggregation 2-fold, whilst also marginally reducing the rate of fragmentation (Table 1). This illustrates that excipients, such as Arg·Glu, can improve the stability of high concentration protein solutions ...Esmail et al. develop and validate a rapid and cost-effective antibody-dependent agglutination (aggregation) test by using SARS-CoV-2 antigen-coated latex particles. The agglutination assay detects SARS-CoV-2 antibodies with 100% specificity and ∼98% sensitivity. They further show that the agglutination assay might be used to …Antibody amyloidogenesis is of great concern for diseases in humans, but has limited application in the development of therapeutic proteins. 23 Moreover, a machine learning-based model that was trained on 21 mAbs was developed to predict therapeutic antibody aggregation rates at 150 mg/ml using structural-based features extracted from MD ...In this set, where we know which antibody is aggregation-prone and classified it as such, the Solubis score clearly outperforms simpler scoring schemes, such as the total raw TANGO score or the number of APRs identified by TANGO in the sequence. For completeness, the Matthews correlation coefficient was 0.83, and the area under curve was 0.93 ...Both antibodies evidenced increasing aggregation rates when approaching extreme temperatures, because of hot and cold denaturation. This behavior was modelled …

It is important to avoid aggregation of monoclonal antibodies during large-scale production for clinical use. In this article, the authors present an approach for establishing maximum permissible hold times for processing of a monoclonal antibody product and highlight the importance of statistical analysis and modelling of the data to …Despite that, the CDR1 domain, predicted to concentrate most of the aggregation-prone residues in both heavy- and light-chain antibody segments, potentially does not have a critical role in FGF2 ...Jul 29, 2016 · Antibody aggregate level is normally controlled in bioprocessing by polishing steps after Protein A capture. This paper studied the Higher Order Structures (HOS) of antibo … Antibody aggregate is a common issue in therapeutic antibodies, which may compromise product efficacy and cause adverse effects. Esmail et al. develop and validate a rapid and cost-effective antibody-dependent agglutination (aggregation) test by using SARS-CoV-2 antigen-coated latex particles. The agglutination assay detects SARS-CoV-2 antibodies with 100% specificity and ∼98% sensitivity. They further show that the agglutination assay might be used to …Aggregate formation is influenced by multiple aspects of the bioproduction process but can be mitigated by good process design and control. Monoclonal antibody (mAb) products have emerged as an extremely important and valuable class of therapeutic products for the treatment of cancer and other diseases such as rheumatoid arthritis, auto ...

The drugs associated with ADCs often have less than desirable aqueous solubility characteristics themselves. These drugs have the propensity to cause inter-antibody aggregation through either exposure of protected aggregation-prone regions within the antibody backbone upon conjugation or through drug–drug mediated interactions . This greatly ...For elution of antibodies captured on a Protein A resin, the pH is shifted from neutral to low pH of 3-3.5, contributing to antibody aggregation to different extents depending on the antibody. To remove these aggregated species, which are classified as impurities that risk eliciting an immunogenic response, IEX is often included further ...Abstract. The size exclusion-high-performance liquid chromatography is a high-throughput analytical method, through isocratic condition, that allows to determine and quantify the level of aggregates and fragments of purified antibodies. Here, we describe the preparation of the samples, chromatographic conditions to be used (buffer …Rabbit recombinant monoclonal Alpha-synuclein aggregate antibody [MJFR-14-6-4-2] - Conformation-Specific. Validated in IHC, Dot, ICC/IF and tested in Mouse, ...Jul 1, 2011 · The phenomenon of protein aggregation is a common issue that compromises the quality, safety, and efficacy of antibodies and can happen at different steps of the manufacturing process, including ...

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Purpose To identify the aggregation mechanism and the stability characteristics of three different monoclonal antibodies under acidic conditions. Methods The aggregation kinetics is analyzed by a combination of light scattering, size exclusion chromatography and fluorescence techniques and the aggregation data are correlated …Protein aggregates may impact significantly the product quality, safety and/or efficacy. This review is intended to summarize four major aspects of protein aggregation – (1) aggregation mechanisms, (2) aggregation-influencing factors, (3) detection of protein aggregates, and (4) control of protein aggregation based on recent literature in ...In addition, aggregation can occur during and after drug fill-finish. Shear stress, agitation, mixing speed and filtration can impact aggregate formation during fill-finish (4, 5). Post fill examples of antibody aggregate formation include product temperature excursions due to improper storage and mishandling such as dropping of vials .Purpose To develop an analytical platform for the estimation as well as characterization of aggregates over the complete size spectrum (from invisible monomer to visible precipitates). Methods Two mAb samples were incubated at 30°C in different buffer systems of protein A chromatography for observing degradation due to aggregation. The aggregation in these samples was quantified by size ...Abstract. Monoclonal antibodies (mAbs) are the fastest-growing biological therapeutics with important applications ranging from cancers, autoimmunity diseases and metabolic …

This study aimed in understanding the degradation behaviors of an IgG 1 subtype therapeutic monoclonal antibody A (mAb-A) associated with pH and buffer species. The information obtained in this study can augment conventional, stability-based screening paradigms by providing the direction necessary for efficient experimental design. …Luan et al. have shown that the ribosomal protein L9 exhibited 76% aggregation at 4 °C and ∼36% aggregation at 25 °C. 109 For antibodies, a decrease in temperature could induce precipitation as seen for the monoclonal IgM cryoglobulin at 14–15 °C. 110 A recent report by Wälchli et al. using two immunoglobulins showed various types and ...Purpose Aggregation of monoclonal antibodies (mAbs) is a common yet poorly understood issue in therapeutic development. There remains a need for high-resolution structural information about conformational changes and intermolecular contacts during antibody aggregation. Methods We used hydrogen exchange mass …Feb 28, 2023 · Antibody-mediated aggregation of the TfR ectodomain was done in a 10 µl reaction containing 10 µM TfR (aa 89–760), 1.5 µM anti-TfR Ab 289, and 0.75 µM anti-human secondary antibody. The calculated aggregation index for each sample is shown in Table 1. Figure 1. UV/Vis Spectra of Antibody Samples. Sample Calculated Aggregation Index Unstressed 1.70 Freeze-thaw stress 52.94 Thermal stress 16.29 Table 1. Calculated aggregation index for each antibody sample. It can be seen from these results that the antibodies in the Protein aggregates may impact significantly the product quality, safety and/or efficacy. This review is intended to summarize four major aspects of protein aggregation – (1) aggregation mechanisms, (2) aggregation-influencing factors, (3) detection of protein aggregates, and (4) control of protein aggregation based on recent literature in ...SEC for Antibody Aggregation Analysis SEC is the standard method for aggregate and fragment analysis of mAbs in biopharmaceutical QC. Aggregates, monomers, dimers, and degradation products of monoclonal antibodies can be separated on size exclusion columns based on their molecular weights under native conditions.We also identified six new spike mutations that mediate antibody resistance, including E554K that threatens SD1 mAbs in clinical development. The BA.2.86 spike also had …undesirable antibody characteristic is the propensity to aggregate. Although there are computational algorithms that predict the propensity of a protein to aggregate from sequence information alone, few consider the relevance of the native structure. The Spatial Aggregation Propensity (SAP) algorithm developed by Chennamsetty et. al. incorporates

Tau protein abnormally aggregates in tauopathies, a diverse group of neurologic diseases that includes Alzheimer’s disease (AD). In early stages of disease, tau becomes hyperphosphorylated and mislocalized, which can contribute to its aggregation and toxicity. We demonstrate that tau phosphorylation at Ser208 (pSer208) promotes microtubule dysfunction …

Trehalose suppresses antibody aggregation during the culture of Chinese hamster ovary cells. Masayoshi Onitsuka, Miki Tatsuzawa, Ryutaro Asano, Izumi Kumagai, ...Antibodies 2016, 5, 19 3 of 23 2. Protein Aggregation 2.1. Why Does a Protein Aggregate? The native protein has the lowest free energy and the most stable conformation [3] (Figure1).Nov 1, 2013 · Self-interaction of a therapeutic monoclonal antibody may cause aggregation, high viscosity, or low solubility, which limits its likelihood of being successfully developed as a drug (i.e., the developability of the drug). 1-5 Ideally, antibodies with self-interaction tendancies could be rapidly eliminated early in the discovery stage, with minimum material consumption, to minimize downstream ... Antibody aggregation also leads to low expression yield from host cells, affecting biological activity, and may increase immunogenic responses [39]. Clearly, such adverse effects must be prevented. Arosio et al. [40] reported that pH and salt concentration are among the main parameters affecting monoclonal antibody aggregation. Low pH (pH < 4.0 ...When it comes to staying informed and up-to-date with the latest news, there are countless options available. One popular choice for many people is Apple News, a news aggregator developed by Apple Inc.Aggregate content in cell culture samples was estimated using a high-resolution size-exclusion chromatography technique, which efficiently resolved the antibody monomer and aggregates in the cell ...Antibody Aggregation Analytical Techniques Aggregation characterization is key to successful biopharmaceutical development and manufacturing. Assessing or analyzing the aggregation level of mAb therapeutic candidates is a regulatory requirement and also an important component of developability assessment.

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Here, we describe a human monoclonal antibody α-IAPP-O selectively targeting IAPP oligomers in T2D. α-IAPP-O neutralizes IAPP aggregate toxicity by preventing membrane disruption and apoptosis ...Abstract. The elicitation of anti-drug antibodies (ADA) against biotherapeutics can have detrimental effects on drug safety, efficacy, and pharmacokinetics. The immunogenicity of biotherapeutics is, therefore, an important issue. There is evidence that protein aggregation can result in enhanced immunogenicity; however, the precise immunological ...Luan et al. have shown that the ribosomal protein L9 exhibited 76% aggregation at 4 °C and ∼36% aggregation at 25 °C. 109 For antibodies, a decrease in temperature could induce precipitation as seen for the monoclonal IgM cryoglobulin at 14–15 °C. 110 A recent report by Wälchli et al. using two immunoglobulins showed various types and ...Jan 25, 2021 · For elution of antibodies captured on a Protein A resin, the pH is shifted from neutral to low pH of 3-3.5, contributing to antibody aggregation to different extents depending on the antibody. To remove these aggregated species, which are classified as impurities that risk eliciting an immunogenic response, IEX is often included further ... The development of stable antibody formulations is challenging, since they have the tendency to be physically and chemically instable in aqueous media. Antibodies are, for example, susceptible to denaturation, aggregation and covalent modifications (2,3). Degradation of antibody formulations due to aggregation phenomena is a particular problem.• The recombinant antibody aggregation in mammalian cell systems is reviewed. • Intracellular environment and extracellular parameters inuence recombinant antibody aggregation . At pH 5.5, both tended to increase the aggregation of an antibody with a strongly positive charge on the Fv, while only NaCl reduced the aggregation of the antibody with a large negative charge patch on the Fv. Arg.HCl reduced the viscosities of antibodies with either a hydrophobicity-driven mechanism or a charge-driven mechanism.Antibody Aggregation Analytical Techniques Aggregation characterization is key to successful biopharmaceutical development and manufacturing. Assessing or analyzing the aggregation …A host of diverse stress techniques was applied to a monoclonal antibody (IgG 2) to yield protein particles with varying attributes and morphologies.Aggregated solutions were evaluated for percent aggregation, particle counts, size distribution, morphology, changes in secondary and tertiary structure, surface hydrophobicity, metal … ….

A host of diverse stress techniques was applied to a monoclonal antibody (IgG 2) to yield protein particles with varying attributes and morphologies.Aggregated solutions were evaluated for percent aggregation, particle counts, size distribution, morphology, changes in secondary and tertiary structure, surface hydrophobicity, metal …Self-interaction of a therapeutic monoclonal antibody may cause aggregation, high viscosity, or low solubility, which limits its likelihood of being successfully developed as a drug (i.e., the developability of the drug). 1-5 Ideally, antibodies with self-interaction tendancies could be rapidly eliminated early in the discovery stage, with minimum material consumption, …Antibody aggregation was analyzed by sephacryl S-300 column (GE healthcare). Solution structure of Ex3-scDb-Fc was assessed by circular dichroism spectroscopy. Results and discussion. Cell culture performance in trehalose containing medium. We cultivated CHO Top-H cell line in 150mM trehalose containing medium. The media osmolalities with and ...Despite that, the CDR1 domain, predicted to concentrate most of the aggregation-prone residues in both heavy- and light-chain antibody segments, potentially does not have a critical role in FGF2 ...The amino acid arginine is frequently used as a solution additive to stabilize proteins against aggregation, especially in the process of protein refolding. Despite arginine's prevalence, the mechanism by which it stabilizes proteins is not presently understood. We propose that arginine deters aggregation by slowing protein−protein association reactions, …On account of the identification of aggregation-prone regions and their contribution to the thermodynamic stability of the protein, predicting and decreasing the aggregation propensity of monoclonal antibodies become available. With years of exploration in antibody engineering, Creative Biolabs has built a full-scale in silico technology platform.Here, we suggest the hypothesis that the heterogeneity of N-linked oligosaccharides is a possible cause of antibody aggregation during the cell culture process.Various studies have investigated the contribution of N-glycosylation to the stability and aggregation of antibodies, as described above.Conversely, the difference in the N …Purpose To develop an analytical platform for the estimation as well as characterization of aggregates over the complete size spectrum (from invisible monomer to visible precipitates). Methods Two mAb samples were incubated at 30°C in different buffer systems of protein A chromatography for observing degradation due to aggregation. The aggregation in these samples was quantified by size ... Antibody aggregation, [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1], [text-1-1]