Water Proton NMR for In Situ Detection of Insulin Aggregates

Taraban, Marc B.; Truong, Huy C.; Feng, Yue; Jouravleva, Elena V.; Анисимов, М. А.; Yu, Yinan

doi:10.1002/jps.24633

Cited by 26 publications

(26 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Furthermore, a pH-induced aggregation procedure was also implemented on human insulin. A non-linear trend between T 2 of water protons and aggregates was observed in this case [113]. The water NMR method was extended to investigate mAb aggregation under various stress conditions, in which the transverse relaxation of water protons detected aggregate formation [114].…”

Section: Nuclear Magnetic Resonance (Nmr) Spectroscopy Applicationmentioning

confidence: 96%

An Evaluation of the Potential of NMR Spectroscopy and Computational Modelling Methods to Inform Biopharmaceutical Formulations

et al. 2018

View full text Add to dashboard Cite

Protein-based therapeutics are considered to be one of the most important classes of pharmaceuticals on the market. The growing need to prolong stability of high protein concentrations in liquid form has proven to be challenging. Therefore, significant effort is being made to design formulations which can enable the storage of these highly concentrated protein therapies for up to 2 years. Currently, the excipient selection approach involves empirical high-throughput screening, but does not reveal details on aggregation mechanisms or the molecular-level effects of the formulations under storage conditions. Computational modelling approaches have the potential to elucidate such mechanisms, and rapidly screen in silico prior to experimental testing. Nuclear Magnetic Resonance (NMR) spectroscopy can also provide complementary insights into excipient–protein interactions. This review will highlight the underpinning principles of molecular modelling and NMR spectroscopy. It will also discuss the advancements in the applications of computational and NMR approaches in investigating excipient–protein interactions.

show abstract

Section: Nuclear Magnetic Resonance (Nmr) Spectroscopy Applicationmentioning

confidence: 96%

An Evaluation of the Potential of NMR Spectroscopy and Computational Modelling Methods to Inform Biopharmaceutical Formulations

et al. 2018

View full text Add to dashboard Cite

show abstract

“…At the same time, the sensitivity of R 2 ( 1 H 2 O) is strongly affected by the size of aggregates and their structural organization, such as formation of water‐accessible cavities inside the assemblies. In the case of rather large aggregates, R 2 ( 1 H 2 O) might respond to their formation even at relatively low total protein concentrations, e.g., <0.5 mg/ml (Taraban et al, ). In general, R 2 ( 1 H 2 O) is most reliably sensitive to protein aggregate content >1% (Taraban, DePaz, et al., ).…”

Section: Strategic Planningmentioning

confidence: 99%

Magnetic Resonance Relaxometry for Determination of Protein Concentration and Aggregation

Taraban

Briggs

2019

CP Protein Science

Self Cite

View full text Add to dashboard Cite

The water‐proton signal, overwhelmingly considered a nuisance in nuclear magnetic resonance spectroscopy, is advantageously used as a tool to assess protein concentration and to detect protein aggregates in aqueous solutions. The protocols in this article describe use of the water‐proton transverse relaxation rate to determine concentration and aggregate content in protein solutions. Detailed recommendations and description of the parameter settings and data processing ensure successful implementation of this technique, even by a user with limited experience in magnetic resonance relaxometry. All measurements are done noninvasively, in a sealed container, without sampling or otherwise aliquoting the solution. The magnetic resonance relaxometry approach offered in this article could be advantageous for analysis of biologics formulations or when use of conventional analytical techniques is not possible. © 2019 by John Wiley & Sons, Inc. Basic Protocol 1: Nuclear magnetic resonance (NMR) relaxometry to measure protein concentration Basic Protocol 2: NMR relaxometry to measure protein aggregation

show abstract

“…Aggregation can be followed by size-exclusion chromatography (SEC) [5] or by sodium dodecyl polyacrylamide gel electrophoresis (SDS-PAGE) [6]. Also, nuclear magnetic resonance (NMR) spectroscopy assays have been suggested for insulin aggregation studies [7,8]. Another option is the application of dynamic light scattering (DLS) methods, which provide information on aggregation [9,10].…”

Section: Introductionmentioning

confidence: 99%

FT-IR versus EC-QCL spectroscopy for biopharmaceutical quality assessment with focus on insulin—total protein assay and secondary structure analysis using attenuated total reflection

Delbeck

Heise

2020

Anal Bioanal Chem

View full text Add to dashboard Cite

For the quality control of biopharmaceutical products, which contain proteins as the most important active ingredients, shelf life may be limited due to inappropriate storage conditions or mechanical stress. For insulins as representatives of life-saving pharmaceuticals, analytical methods are needed, which are providing additional information than obtained by assays for total protein quantification. Despite sophisticated formulations, the chemical stability may be challenged by temperatures deviating from recommended conditions or shear rate exposure under storage, leading to misfolding, nucleation, and subsequent fibril formation, accompanied by a decrease in bioactivity. A reliable method for insulin quantification and determination of secondary structure changes has been developed by attenuated total reflection (ATR) Fourier-transform infrared spectroscopy of insulin formulations by a silver halide fiber-coupled diamond probe with subsequent dry-film preparation. A special emphasis has been placed on the protein amide I band evaluation, for which spectral band analysis provides unique information on secondary structure fractions for intact and misfolded insulins. Quantitative measurements are possible down to concentrations of less than 0.5 mg/ml, whereas the dry-film preparation delivers high signal-to-noise ratios due to the prior water evaporation, thus allowing a reliable determination of secondary structure information.

show abstract

Water Proton NMR for In Situ Detection of Insulin Aggregates

Cited by 26 publications

References 35 publications

An Evaluation of the Potential of NMR Spectroscopy and Computational Modelling Methods to Inform Biopharmaceutical Formulations

An Evaluation of the Potential of NMR Spectroscopy and Computational Modelling Methods to Inform Biopharmaceutical Formulations

Magnetic Resonance Relaxometry for Determination of Protein Concentration and Aggregation

FT-IR versus EC-QCL spectroscopy for biopharmaceutical quality assessment with focus on insulin—total protein assay and secondary structure analysis using attenuated total reflection

Contact Info

Product

Resources

About