Analytical Tools for Physicochemical Characterization and Fingerprinting

Keire, David A.

doi:10.1007/978-3-030-11751-1_6

Cited by 2 publications

(2 citation statements)

References 76 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…With the development of higher field strength magnets and cryogenic probes, modern high-resolution NMR spectroscopy is a non-invasive and sensitive method for protein molecular structure characterization [ 14 , 15 , 16 , 17 , 18 ]. However, several assumptions among stakeholders have limited the application of NMR on formulated DPs.…”

Section: Introductionmentioning

confidence: 99%

NMR Spectroscopy for Protein Higher Order Structure Similarity Assessment in Formulated Drug Products

et al. 2021

Self Cite

View full text Add to dashboard Cite

Peptide and protein drug molecules fold into higher order structures (HOS) in formulation and these folded structures are often critical for drug efficacy and safety. Generic or biosimilar drug products (DPs) need to show similar HOS to the reference product. The solution NMR spectroscopy is a non-invasive, chemically and structurally specific analytical method that is ideal for characterizing protein therapeutics in formulation. However, only limited NMR studies have been performed directly on marketed DPs and questions remain on how to quantitively define similarity. Here, NMR spectra were collected on marketed peptide and protein DPs, including calcitonin-salmon, liraglutide, teriparatide, exenatide, insulin glargine and rituximab. The 1D 1H spectral pattern readily revealed protein HOS heterogeneity, exchange and oligomerization in the different formulations. Principal component analysis (PCA) applied to two rituximab DPs showed consistent results with the previously demonstrated similarity metrics of Mahalanobis distance (DM) of 3.3. The 2D 1H-13C HSQC spectral comparison of insulin glargine DPs provided similarity metrics for chemical shift difference (Δδ) and methyl peak profile, i.e., 4 ppb for 1H, 15 ppb for 13C and 98% peaks with equivalent peak height. Finally, 2D 1H-15N sofast HMQC was demonstrated as a sensitive method for comparison of small protein HOS. The application of NMR procedures and chemometric analysis on therapeutic proteins offer quantitative similarity assessments of DPs with practically achievable similarity metrics.

show abstract

Section: Introductionmentioning

confidence: 99%

NMR Spectroscopy for Protein Higher Order Structure Similarity Assessment in Formulated Drug Products

et al. 2021

Self Cite

View full text Add to dashboard Cite

show abstract

“…Optimisation of formulation conditions, and continued assessment of formulation stability, requires analytical techniques capable of assessing both protein and small molecule content and behavior, ideally in situ in intact formulations. However, in practice, a wide range of techniques is deployed, − and these techniques typically require manipulation of high-concentration formulations, such as dilution, addition of a probe molecule, or salt removal, potentially leading to changes in protein and small molecule structure and behavior.…”

Section: Introductionmentioning

confidence: 99%

Comprehensive Assessment of Protein and Excipient Stability in Biopharmaceutical Formulations Using ¹H NMR Spectroscopy

Bramham

Podmore

Davies

et al. 2020

ACS Pharmacol. Transl. Sci.

View full text Add to dashboard Cite

Biopharmaceutical proteins are important drug therapies in the treatment of a range of diseases. Proteins, such as antibodies (Abs) and peptides, are prone to chemical and physical degradation, particularly at the high concentrations currently sought for subcutaneous injections, and so formulation conditions, including buffers and excipients, must be optimized to minimize such instabilities. Therefore, both the protein and small molecule content of biopharmaceutical formulations and their stability are critical to a treatment’s success. However, assessing all aspects of protein and small molecule stability currently requires a large number of analytical techniques, most of which involve sample dilution or other manipulations which may themselves distort sample behavior. Here, we demonstrate the application of 1 H nuclear magnetic resonance (NMR) spectroscopy to study both protein and small molecule content and stability in situ in high-concentration (100 mg/mL) Ab formulations. We show that protein degradation (aggregation or fragmentation) can be detected as changes in 1D 1 H NMR signal intensity, while apparent relaxation rates are specifically sensitive to Ab fragmentation. Simultaneously, relaxation-filtered spectra reveal the presence and degradation of small molecule components such as excipients, as well as changes in general solution properties, such as pH. 1 H NMR spectroscopy can thus provide a holistic overview of biopharmaceutical formulation content and stability, providing a preliminary characterization of degradation and acting as a triaging step to guide further analytical techniques.

show abstract

Analytical Tools for Physicochemical Characterization and Fingerprinting

Cited by 2 publications

References 76 publications

NMR Spectroscopy for Protein Higher Order Structure Similarity Assessment in Formulated Drug Products

NMR Spectroscopy for Protein Higher Order Structure Similarity Assessment in Formulated Drug Products

Comprehensive Assessment of Protein and Excipient Stability in Biopharmaceutical Formulations Using ¹H NMR Spectroscopy

Contact Info

Product

Resources

About

Analytical Tools for Physicochemical Characterization and Fingerprinting

Cited by 2 publications

References 76 publications

NMR Spectroscopy for Protein Higher Order Structure Similarity Assessment in Formulated Drug Products

NMR Spectroscopy for Protein Higher Order Structure Similarity Assessment in Formulated Drug Products

Comprehensive Assessment of Protein and Excipient Stability in Biopharmaceutical Formulations Using 1H NMR Spectroscopy

Contact Info

Product

Resources

About

Comprehensive Assessment of Protein and Excipient Stability in Biopharmaceutical Formulations Using ¹H NMR Spectroscopy