Rapid Conformational Analysis of Protein Drugs in Formulation by Hydrogen/Deuterium Exchange Mass Spectrometry

Nazari, Zeinab; Weert, Marco van de; Bou-Assaf, George M.; Houde, Damian J.; Weiskopf, Andrew; Rand, Kasper D.

doi:10.1016/j.xphs.2016.07.006

Cited by 13 publications

(15 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the last 5 years, there has been methodological advances w.r.t range of temperature required for HDX experiments, method reproducibility, significance testing, and application of HDX-MS for similarity assessments. This includes the optimization of HDX-MS methodology based on manual solid-phase extraction to allow a fast and simplified conformational analysis of proteins under pharmaceutically relevant formulation conditions as demonstrated on interferon-β-1a in-formulation samples (E. Nazari et al, 2016 ) ( Supplementary Table S7 ).…”

Section: Global Landscape On Biosimilar Approvalsmentioning

confidence: 99%

Analytical Similarity Assessment of Biosimilars: Global Regulatory Landscape, Recent Studies and Major Advancements in Orthogonal Platforms

Nupur

Joshi

Gulliarme

et al. 2022

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Biopharmaceuticals are one of the fastest-growing sectors in the biotechnology industry. Within the umbrella of biopharmaceuticals, the biosimilar segment is expanding with currently over 200 approved biosimilars, globally. The key step towards achieving a successful biosimilar approval is to establish analytical and clinical biosimilarity with the innovator. The objective of an analytical biosimilarity study is to demonstrate a highly similar profile with respect to variations in critical quality attributes (CQAs) of the biosimilar product, and these variations must lie within the range set by the innovator. This comprises a detailed comparative structural and functional characterization using appropriate, validated analytical methods to fingerprint the molecule and helps reduce the economic burden towards regulatory requirement of extensive preclinical/clinical similarity data, thus making biotechnological drugs more affordable. In the last decade, biosimilar manufacturing and associated regulations have become more established, leading to numerous approvals. Biosimilarity assessment exercises conducted towards approval are also published more frequently in the public domain. Consequently, some technical advancements in analytical sciences have also percolated to applications in analytical biosimilarity assessment. Keeping this in mind, this review aims at providing a holistic view of progresses in biosimilar analysis and approval. In this review, we have summarized the major developments in the global regulatory landscape with respect to biosimilar approvals and also catalogued biosimilarity assessment studies for recombinant DNA products available in the public domain. We have also covered recent advancements in analytical methods, orthogonal techniques, and platforms for biosimilar characterization, since 2015. The review specifically aims to serve as a comprehensive catalog for published biosimilarity assessment studies with details on analytical platform used and critical quality attributes (CQAs) covered for multiple biotherapeutic products. Through this compilation, the emergent evolution of techniques with respect to each CQA has also been charted and discussed. Lastly, the information resource of published biosimilarity assessment studies, created during literature search is anticipated to serve as a helpful reference for biopharmaceutical scientists and biosimilar developers.

show abstract

Section: Global Landscape On Biosimilar Approvalsmentioning

confidence: 99%

Analytical Similarity Assessment of Biosimilars: Global Regulatory Landscape, Recent Studies and Major Advancements in Orthogonal Platforms

Nupur

Joshi

Gulliarme

et al. 2022

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

show abstract

“…For the characterization of the intact MGL, 30 pmol of protein was incubated for 45 min at rt in 20 mM Tris buffer, pH 8.0 (final incubation volume: 50 μL) either in the absence or in the presence of a 4:1 molar excess of compound 13. At the end of the incubation time, intact protein samples were injected in a rapid desalting system constituted of a Waters Acquity HPLC (Waters Corp., Milford, MA) with an injection loop containing a Valco, a 6port injection valve 91 (VICI AG International, Schenkon, Switzerland) equipped with a microcolumn 92 (approximately 2 μL bed volume of C4 Poros 20 R2), and proteins were eluted into an electrospray ion source at a flow rate of 40 μL/min employing an 8.5 min gradient going from 10 to 90% acetonitrile in 0.1% formic acid. Positive ion-electrospray ionization mass spectra of eluted proteins were acquired on a Waters Synapt G2 HDMS mass spectrometer (Waters Corp., Milford, MA).…”

Section: ■ Experimental Sectionmentioning

confidence: 99%

Benzisothiazolinone Derivatives as Potent Allosteric Monoacylglycerol Lipase Inhibitors That Functionally Mimic Sulfenylation of Regulatory Cysteines

et al. 2019

View full text Add to dashboard Cite

We describe a set of benzisothiazolinone (BTZ) derivatives that are potent inhibitors of monoacylglycerol lipase (MGL), the primary degrading enzyme for the endocannabinoid 2-arachidonoyl-sn-glycerol (2-AG). Structure–activity relationship studies evaluated various substitutions on the nitrogen atom and the benzene ring of the BTZ nucleus. Optimized derivatives with nanomolar potency allowed us to investigate the mechanism of MGL inhibition. Site-directed mutagenesis and mass spectrometry experiments showed that BTZs interact in a covalent reversible manner with regulatory cysteines, Cys201 and Cys208, causing a reversible sulfenylation known to modulate MGL activity. Metadynamics simulations revealed that BTZ adducts favor a closed conformation of MGL that occludes substrate recruitment. The BTZ derivative 13 protected neuronal cells from oxidative stimuli and increased 2-AG levels in the mouse brain. The results identify Cys201 and Cys208 as key regulators of MGL function and point to the BTZ scaffold as a useful starting point for the discovery of allosteric MGL inhibitors.

show abstract

“…5 The increasing use of HDX-MS has been facilitated by rapid advances in the hardware and software that collect and analyze HDX-MS kinetics measurements. In short, HDX-MS has emerged from the "quicksand" of frontier science, 6 and it has become a mainstay tool in modern pharmaceutical 4,[7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26] and structural biology laboratories. 3,23,[27][28][29][30][31][32][33][34][35] Major advantages of HDX-MS for characterizing the conformational dynamics of proteins are its availability to make measurements under physiological conditions, its modest sample requirements, its use of proteomic informatics that link each HDX-MS peptide sequence directly to a portion of the subject protein, and its nearly unlimited capability to characterize large proteins, for example, antibodies (~ 150 kBa) 10,12,16,19,35 and viral capsids (~2 MDa).…”

Section: Graphical Abstract Introductionmentioning

confidence: 99%

“…The increasing use of HDX-MS has been facilitated by rapid advances in the hardware and software that collect and analyze HDX-MS kinetics measurements. In short, HDX-MS has emerged from the “quicksand” of frontier science, and it has become a mainstay tool in modern pharmaceutical ,− and structural biology laboratories. ,,− …”

Section: Introductionmentioning

confidence: 99%

Interlaboratory Comparison of Hydrogen–Deuterium Exchange Mass Spectrometry Measurements of the Fab Fragment of NISTmAb

et al. 2019

View full text Add to dashboard Cite

Descriptions of materials, metrological methods, computational methods, and supplementary results. Figures of HDX-MS publications and citations versus publication year, histogram of peptide sequence lengths, sequence coverage maps, performance of instrumentsoftware configurations, repeatability plots, %E corrected peptide t HDX versus log 10 (t HDX ) for eight peptides. Tables of instrumentation,software, peptide search methodology, and operating conditions of proteolytic, chromatographic components, and effects of peptide charge on deuterium uptake (PDF)

show abstract

Rapid Conformational Analysis of Protein Drugs in Formulation by Hydrogen/Deuterium Exchange Mass Spectrometry

Cited by 13 publications

References 35 publications

Analytical Similarity Assessment of Biosimilars: Global Regulatory Landscape, Recent Studies and Major Advancements in Orthogonal Platforms

Analytical Similarity Assessment of Biosimilars: Global Regulatory Landscape, Recent Studies and Major Advancements in Orthogonal Platforms

Benzisothiazolinone Derivatives as Potent Allosteric Monoacylglycerol Lipase Inhibitors That Functionally Mimic Sulfenylation of Regulatory Cysteines

Interlaboratory Comparison of Hydrogen–Deuterium Exchange Mass Spectrometry Measurements of the Fab Fragment of NISTmAb

Contact Info

Product

Resources

About