High-Resolution NMR Services
In the complex world of biopharmaceutical development, knowing your protein's exact structure is critical for early-stage R&D and candidate selection. Our High-Resolution Solution Nuclear Magnetic Resonance (NMR) service allows biologics to be observed in their native, liquid state.
We provide atomic-level structural fingerprints to support structural-equivalence assessment, formulation optimization, interaction mapping, and early-stage candidate selection.
- Characterize protein higher-order structure in solution.
- Evaluate structural dynamics, flexibility, and formulation-dependent changes.
- Map ligand, drug, epitope, and protein-interaction sites with high resolution.
In line with ICH Q6B Guidance for biologic characterization
Jump to Section
- Why High-Resolution NMR Matters
- Service Capabilities
- Biologic Modalities
- Analytical Approaches
- Service Advantages
- Instrument Platform
- Analysis Workflow
- Report Deliverables
- FAQs
- Sample Requirements
Why High-Resolution NMR Matters in Biologic Development
The performance of a biologic depends not only on its primary sequence, but also on how the molecule folds, fluctuates, and responds to its solution environment. Small local changes in conformation or dynamics may affect stability, molecular recognition, aggregation behavior, or biological activity, even when the overall protein appears structurally similar by lower-resolution methods.
High-resolution solution NMR provides detailed structural fingerprints while the protein remains in solution. Resonance positions, signal intensities, and line shapes reflect local chemical environments and molecular motions. Comparing these spectral features across candidates, batches, formulations, or reference materials can reveal subtle structural differences and, when appropriate data are available, help localize the affected molecular regions.
This sensitivity makes NMR valuable for candidate selection, formulation screening, stability investigations, interaction studies, and higher-order structure comparability. It can also evaluate how buffer composition, pH, temperature, excipients, or storage conditions influence a biologic's structure and dynamics.
When combined with Circular Dichroism, FTIR, and Differential Scanning Calorimetry, high-resolution NMR adds localized—and in suitable studies, residue-level—information, providing a more complete understanding of biologic structure, stability, and comparability.
Fit-for-purpose study design. The appropriate experiment depends on molecular size, concentration, spectral complexity, labeling status, formulation background, and whether the objective is fingerprinting, residue-level mapping, or full structure determination.
High-Resolution NMR Service Capabilities
Higher-Order Structure Analysis
Confirm secondary and tertiary structure with detailed 1D and 2D NMR mapping to assess whether the protein is correctly folded and structurally consistent.
Protein Dynamics and Flexibility
Assess molecular flexibility and solution-state behavior to understand structural dynamics relevant to function and developability.
Formulation and Stability Testing
Evaluate how buffers, pH, temperature, and formulation components influence structure, unfolding, degradation, or aggregation risk.
Binding and Interaction Mapping
Map epitopes and identify regions involved in ligand, drug, or protein interactions to support mechanistic research and rational design.
High-Resolution NMR Across Biologic Modalities
We tailor NMR strategy to the molecule, formulation environment, and required level of structural detail.
- Monoclonal Antibodies and Biosimilars
Compare spectral fingerprints with reference materials and investigate HOS consistency, domain-level changes, or formulation-dependent differences. - Recombinant Proteins and Peptides
Characterize folding, dynamics, conformational integrity, and—when feasible—residue assignments or three-dimensional structures. - Protein–Ligand and Protein–Protein Complexes
Map binding interfaces, epitopes, and chemical-shift perturbations associated with molecular recognition. - Formulated Biologics and Candidate Panels
Support formulation scientists and discovery teams by comparing structural fingerprints across buffers, stress conditions, variants, or candidate molecules.
Analytical Approaches for High-Resolution NMR Characterization
The study can range from rapid spectral profiling to multidimensional structure determination. Final experiment selection is confirmed during feasibility review.
| Approach | Best suited for | Typical considerations and outputs |
| 1D NMR profiling | Rapid HOS fingerprinting, screening, and sample-quality assessment | Often suitable for unlabeled samples. Provides spectral fingerprints and comparative profiles with a typical turnaround of approximately two weeks. |
| 2D NMR mapping | Higher-resolution fingerprinting, comparability, and interaction mapping | Experiment and labeling needs depend on molecular size and target nuclei. Outputs can include spectral overlays, chemical-shift changes, and mapped features. |
| Multidimensional / 3D NMR | Resonance assignment and solution-structure calculation for suitable proteins | Generally requires appropriately isotope-labeled material, commonly 15N or 15N/13C. Outputs may include assignments, structure ensembles, and coordinates. |
Service Advantages
Our high-resolution NMR service combines native-state observation with detailed structural evidence for early-stage research decisions.
- Native-State Analysis
Observe biologics in solution under near-physiological or project-relevant conditions to reflect native behavior as closely as practical. - Data-Driven R&D
Use atomic-level structural evidence to guide candidate selection, formulation screening, and early-pipeline decisions. - Proactive Risk Mitigation
Detect subtle structural shifts, misfolding, heterogeneity, or formulation effects during discovery and screening. - Non-Destructive Analysis
NMR is generally non-destructive under suitable native conditions, supporting sample recovery when stability and acquisition conditions allow.
Instrument Platform for High-Resolution NMR Analysis
Creative Proteomics uses high-field NMR spectroscopy with experiment-specific pulse sequences and acquisition conditions. High-sensitivity probe technology, including cryogenic probes where appropriate, may support data acquisition at lower concentrations. Platform configuration is selected according to protein size, labeling status, sample volume, concentration, formulation background, and study objective.
Sample sensitivity. Although NMR is generally non-destructive, prolonged acquisition at high concentration may affect sensitive proteins. Sample recovery and post-analysis condition should be discussed during project planning.
High-Resolution NMR Analysis Workflow
To support high-quality data, we follow a seven-stage workflow for high-resolution solution NMR analysis:
- 1
Project Consultation & Feasibility
Understand your goals and sample requirements to determine the best NMR strategy.
- 2
Sample Preparation & Quality Control
Prepare high-quality samples with careful attention to protein expression, purity, and homogeneity to ensure suitability for NMR analysis.
- 3
NMR Data Acquisition
Use advanced NMR instruments to gather high-resolution data. This includes high-field spectroscopy and optimizing conditions for each sample.
- 4
Data Processing & Resonance Assignment
Process the raw NMR data and assign chemical shifts to identify key spectral features, including backbone and side-chain resonances.
- 5
Structure Calculation & Refinement
Where applicable, calculate a 3D structure from the NMR data and refine the model using computational tools, ensuring accuracy and precision.
- 6
Structure Validation & Analysis
Validate the resulting structure using tools like MolProbity. Perform analysis of structural dynamics and functional mappings.
- 7
Reporting & Data Delivery
Provide a comprehensive report with structural insights, coordinates (PDB), and processed NMR data, supporting downstream decision-making and further analysis.
Isotope-labeling requirement. Three-dimensional structure calculation and refinement generally require appropriately isotope-labeled samples, such as 15N or 15N/13C. Unlabeled samples used for fit-for-purpose 1D or 2D analyses may proceed directly from data interpretation to reporting.
Report Deliverables
Depending on the selected NMR study, deliverables can include:
- A final report describing sample conditions, acquisition strategy, results, and interpretation.
- Processed NMR spectra and project-relevant spectral overlays or comparative outputs.
- Chemical-shift lists or resonance assignments when included in the project scope.
- A three-dimensional structure ensemble and coordinate files in PDB format, where applicable.
- Processed NMR data suitable for downstream review or deposition planning, as agreed.
Frequently Asked Questions About High-Resolution NMR
How does NMR compare with Circular Dichroism or FTIR?
CD and FTIR provide broad views of overall secondary structure, such as estimated alpha-helix and beta-sheet content. High-resolution NMR provides more localized spectral information and can identify which residues or regions are affected by a structural or interaction-related change when the experiment supports assignment.
Can these data be used for regulatory submissions?
Our High-Resolution NMR services are for Research Use Only. The data are intended to support early-stage discovery, clone or candidate selection, formulation screening, biosimilar research, and internal R&D decisions.
What if my protein aggregates at the high concentrations required for NMR?
Our scientists can evaluate buffer optimization, pH adjustment, alternative sample volumes, and suitable pulse sequences or probe technology. Feasibility still depends on the molecule's stability and the sensitivity required for the selected experiment.
What is the typical turnaround time?
One-dimensional profiling typically takes approximately two weeks, while more extensive 2D or 3D structural studies typically take approximately 4–6 weeks.
Sample Requirements for High-Resolution NMR Analysis
Please contact our technical team for tailored preparation instructions. General planning guidelines are summarized below.
| Parameter | General guideline |
| Purity and homogeneity | Samples should ideally be >95% pure, as assessed by an appropriate method such as SEC or SDS-PAGE, to reduce overlapping signals from impurities. |
| Buffer composition | Provide the complete buffer formulation. Strong protonated buffer or excipient signals can interfere with some experiments; deuterated alternatives or buffer exchange may be recommended. Avoid paramagnetic salts when possible. |
| D₂O | An addition of approximately 5–10% D₂O is commonly required for the NMR lock signal, subject to the selected experiment and sample stability. |
| Isotope labeling | High-resolution multidimensional mapping and structure calculation commonly require 15N or 15N/13C labeling. Some small-protein, 1D, or 2D studies may use unlabeled material. |
| Concentration | Original planning ranges: approximately 1–5 mg/mL for 1D HOS profiling and around 10 mg/mL for extensive 2D/3D mapping; final requirements depend on molecular weight and experiment sensitivity. |
| Volume | Typical volumes are approximately 150–200 µL for 3 mm microprobes or 35–45 µL for ultra-micro 1.7 mm probes, subject to platform availability and project design. |
| Shipping | Ship on dry ice or cold packs according to the protein's stability profile and include the completed Sample Submission Form. |
Before shipping samples. Confirm concentration, volume, buffer compatibility, D₂O content, isotope labeling, storage conditions, and the required experiment with our technical team.
References
- Arbogast LW, Brinson RG, Formolo T, Hoopes JT, Marino JP. 2D 1H(N), 15N Correlated NMR Methods at Natural Abundance for Obtaining Structural Maps and Statistical Comparability of Monoclonal Antibodies. Pharmaceutical Research. 2016;33:462–475. doi:10.1007/s11095-015-1802-3.
- Brinson RG, Marino JP, Delaglio F, et al. Enabling adoption of 2D-NMR for the higher order structure assessment of monoclonal antibody therapeutics. mAbs. 2019;11:94–105. doi:10.1080/19420862.2018.1544454.
- Aubin Y, Hodgson DJ, Thach WB, Gingras G, Sauvé S. Monitoring Effects of Excipients, Formulation Parameters and Mutations on the High Order Structure of Filgrastim by NMR. Pharmaceutical Research. 2015;32:3365–3375. doi:10.1007/s11095-015-1713-3.
