Comparative Stability Studies of Different Infliximab and Biosimilar CT-P13 Clinical Solutions by Combined Use of Physicochemical Analytical Techniques and Enzyme-Linked Immunosorbent Assay (ELISA)

Hermosilla, Jesús; Martín, R. Sánchez; Pérez-Robles, Raquel; Salmerón-García, Antonio; Casares, Salvador; Cabeza, José; Cuadros-Rodrı́guez, Luis; Navas, Natalia

doi:10.1007/s40259-019-00342-9

Cited by 14 publications

(18 citation statements)

References 20 publications

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“…No meaningful clinical differences have been reported to date. CT‐P13 also demonstrated high levels of similarity to infliximab 32 . Differences in the native isoform MS profile were seen between CT‐P13 and infliximab, which were expected based on different glycosylation patterns.…”

Section: Discussionmentioning

confidence: 86%

Physicochemical analysis and biological characterization of FKB327 as a biosimilar to adalimumab

Schreiber

Yamamoto

Muniz

et al. 2020

Pharmacology Res & Perspec

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FKB327 was approved by the European Medicines Agency as a biosimilar to European‐authorized adalimumab (Humira®; AbbVie Inc). Adalimumab is a monoclonal antibody, binding and inhibiting tumor necrosis factor (TNF)‐α with use indicated for several immune‐mediated, chronic, and inflammatory disorders. The approval is based on high similarity in the physicochemical properties between FKB327 and adalimumab. The objective of this study is to assess the biological similarity, with regard to Fab‐ and Fc‐associated functions, and describe the relationship between physicochemical and biological characterization and functional activity. State‐of‐the‐art orthogonal techniques were implemented to assess the structure and function of FKB327. Peptide mapping with liquid chromatography and mass spectrometry, capillary electrophoresis–sodium dodecyl sulfate, ultraviolet circular dichroism, size‐exclusion high‐performance liquid chromatography (HPLC), and cation exchange HPLC were the techniques used to assess structure. Functional activity was assessed with enzyme‐linked immunosorbent assay, surface plasmon resonance, and cell‐based assays. The polypeptide sequence of FKB327 was identical to that of adalimumab. FKB327 also was demonstrated to have a similar secondary and tertiary structure to adalimumab. Posttranslational heterogeneities, along with size and charge variants, were not clinically meaningful. FKB327 binds to TNF‐α, FcγR, the neonatal Fc receptor, and C1q, and induces apoptosis, antibody‐dependent cellular cytotoxicity, and complement‐dependent cytotoxicity. The binding and activity of FKB327 were similar to that of adalimumab. FKB327 shares similar structure and activity with adalimumab. Based on characterization of physicochemical and biological properties, FKB327 is expected to have a similar safety, immunogenicity, and efficacy profile to adalimumab.

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Section: Discussionmentioning

confidence: 86%

Physicochemical analysis and biological characterization of FKB327 as a biosimilar to adalimumab

Schreiber

Yamamoto

Muniz

et al. 2020

Pharmacology Res & Perspec

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“…The experimental conditions were similar to those used in ref. 32 . Measurements were carried out on a Cary Eclipse spectrofluorometer (Agilent, Santa Clara, CA, USA) equipped with a Peltier system for temperature control.…”

Section: Methodsmentioning

confidence: 99%

“…The experimental conditions were similar to those used in ref. 32 . DLS measurements were carried out in a Protein Solutions DynaPro-99 System Dynamic Light Scattering Module equipped with a Temperature Control Micro Sampler (Wyatt, Santa Bárbara, California, USA).…”

Section: Methodsmentioning

confidence: 99%

“…Functional-based method: enzyme-linked immunosorbent assay (ELISA). This method was here optimized for ziv-AFL, starting from the outline of that optimized for infliximab 32 . An indirect and non-competitive ELISA based on the specific interaction of ziv-AFL with VEGF was carried out to test the biological activity of ziv-AFL after the various stress factors had been applied.…”

Section: Size Exclusion Chromatography (Se/hplc-dad)mentioning

confidence: 99%

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Comprehensive biophysical and functional study of ziv-aflibercept: characterization and forced degradation

Hermosilla

Pérez-Robles

Salmerón-García

et al. 2020

Sci Rep

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Bones 4 & natalia navas 1* Aflibercept (AFL) is an Fc fusion protein used in the treatment of colorectal cancers and different ophthalmological diseases. There are two medicines in which AFL is the active substance: Zaltrap and Eylea, referred as ziv-AFL and AFL respectively. No proper accelerated degradation studies were published on either AFL or ziv-AFL. These studies are essential during research, development and manufacturing stages. Here, we characterized ziv-AFL and submitted it to different stress conditions: light, 60 °C, freeze-thaw cycles, changes in pH, high hypertonic solution and strong denaturing conditions. We used an array of techniques to detect aggregation (SE-HPLC/DAD and DLS), changes in secondary structure (Far-UV circular dichroism), changes in conformation or tertiary structure (Intrinsic tryptophan fluorescence) and alterations in functionality (ELISA). Results indicate that aggregation is common degradation pathway. Two different types of aggregates were detected: dimers and high molecular weight aggregates attributed to β-amyloid-like structures. Secondary structure was maintained in most of the stress tests, while conformation was altered by almost all the tests except for the freeze-thaw cycles. Functionality, evaluated by its immunochemical reaction with VEGF, was found to be stable but with decrease when exposed to light and with likely partial inactivation of the drug when pH was altered. Fc-fusions proteins are a well-established class of biotherapeutics. They are composed of the Fc region of the IgG antibody (Hinge-CH2-CH3) and a desired linked protein. Although the top-selling biotherapeutics are monoclonal antibodies (mAbs), there is increasing use of therapeutic Fc-fusion proteins in medicine today 1. One of these is aflibercept (AFL), a recombinant human Fc-fusion protein that-as indicated in the assessment report of Zaltrap of the European Medicine Agency 2-"acts as a high-affinity soluble decoy receptor that preferentially binds to vascular endothelial growth factor A (VEGF-A), VEGF-B and placenta growth factor (PlGF) and prevents these factors from activating their endogenous receptors. By blocking this pathway, AFL exerts direct anti-cancer activity and boosts the anti-cancer activity of chemotherapy agents by preventing new tumour vessel growth, regressing existing tumour vessels, normalizing vasculature, negatively affecting tumour cell function, offsetting the effects of chemotherapy induction of VEGF levels and potentially inhibiting VEGF repression of dendritic cell function" 2. As a VEGF inhibitor, AFL is also classified as a VEGF-trap together with bevacizumab and ranibizumab. Cancer therapy is not the only field of medicine to benefit from AFL, which is also being used to treat several ophthalmological diseases characterized by neovascularization. As the administration to patient varies depending on the particular illness they are suffering, two different medicines are currently available, Zaltrap (Sanofi-Aventis US, LLC, Bridgewater, New Jersey, USA and Regeneron...

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“…By peptide mapping they showed methionine oxidations that were comparable between the reference product and the biosimilar [29]. Some studies have applied forced oxidation as well as other forced degradation treatments on bevacizumab and infliximab for analysis of aggregate formation, for binding assays and to support method validation [30,31,32,33]. However, as these studies were conducted on the intact mAbs, no structural information about PTMs introduced by forced degradation was provided.…”

Section: Introductionmentioning

confidence: 99%

Forced Degradation Testing as Complementary Tool for Biosimilarity Assessment

et al. 2019

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Oxidation of monoclonal antibodies (mAbs) can impact their efficacy and may therefore represent critical quality attributes (CQA) that require evaluation. To complement classical CQA, bevacizumab and infliximab were subjected to oxidative stress by H2O2 for 24, 48, or 72 h to probe their oxidation susceptibility. For investigation, a middle-up approach was used utilizing liquid chromatography hyphenated with mass spectrometry (LC-QTOF-MS). In both mAbs, the Fc/2 subunit was completely oxidized. Additional oxidations were found in the light chain (LC) and in the Fd’ subunit of infliximab, but not in bevacizumab. By direct comparison of methionine positions, the oxidized residues in infliximab were assigned to M55 in LC and M18 in Fd’. The forced oxidation approach was further exploited for comparison of respective biosimilar products. Both for bevacizumab and infliximab, comparison of posttranslational modification profiles demonstrated high similarity of the unstressed reference product (RP) and the biosimilar (BS). However, for bevacizumab, comparison after forced oxidation revealed a higher susceptibility of the BS compared to the RP. It may thus be considered a useful tool for biopharmaceutical engineering, biosimilarity assessment, as well as for quality control of protein drugs.

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Comparative Stability Studies of Different Infliximab and Biosimilar CT-P13 Clinical Solutions by Combined Use of Physicochemical Analytical Techniques and Enzyme-Linked Immunosorbent Assay (ELISA)

Cited by 14 publications

References 20 publications

Physicochemical analysis and biological characterization of FKB327 as a biosimilar to adalimumab

Physicochemical analysis and biological characterization of FKB327 as a biosimilar to adalimumab

Comprehensive biophysical and functional study of ziv-aflibercept: characterization and forced degradation

Forced Degradation Testing as Complementary Tool for Biosimilarity Assessment

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