Analytical chemistry : NMR, 2D NMR, IRMS

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Our experts in analytical chemistry provide your projects with exceptional technical resources and know-how.

Our Capacités experts in analytical chemistry have developed an exceptional know-how in Nuclear Magnetic Resonance (1D NMR and 2D NMR) and in Isotope Ratio Mass Spectrometry (IRMS).
Access to a large pool of equipment allows us to support industrial projects, from sample preparation to characterization.

We are specialized in very high precision quantification projects and in the analysis of complex matrices. We help our clients to innovate in the study of metabolism, the fight against counterfeiting, the determination of the purity of active molecules, or the monitoring of chemical transformations.

Summary

Services

Strategy and consulting

Diagnosis and selection

Process development

Quality control

Development of methods

Metabolomic and lipidomic analyses

Validate a therapeutic target or refine the description of a pathology through the identification and qualification of biomarkers responsible for changes between two states.

Our experts master different types of analyses of biological samples. They perform targeted and non-targeted profiling and highlight changes between states of the biological matrices studied.

In a nutshell

Targeted analysis

Metabolomic profiling

Metabolic Footprint

Analysis of matrices

Data processing

Targeted analysis

  • Structural elucidation by screening of unknowns: NMR, mass spectrometry (low and/or high resolution), mainly in LC-MS: HPLC-MSn base resolution, HPLC-DAD-MSn high resolution, HPLC-UV, HPLC-FLR, GS-MS, UV-vis and NIR
  • On-line monitoring of chemical and biochemical transformations by benchtop NMR
  • Dosing : pigment by spectrometry, proteins by colorimetric method, total bones by colorimetric method
  • Performing statistical analyses such as PCA and PLS-DA

Metabolomic profiling

  • Compound profiling: polar by GCMS and water-soluble by HPLC
  • Class profiling by HPTLC
  • Family profiling by FTIR
  • One dimensional (1D) and/or two dimensional (2D) NMR profiling of biological matrices. Targeted or non-targeted analyses

Metabolic Footprint

  • Possible biomarkers identification
  • Targeted quantification of key metabolites in biological samples

Analysis of matrices

  • Metabolic or lipidomic extracts obtained from animals or plant matrices, bacteria, etc.
  • Biofluids: urine, blood, plasma, culture supernatants, etc.
  • Validation of regulatory status for commercial pure molecules
  • Synthesized or manufactured molecules not available on the market
  • Internal reference substances

Data processing

  • Pre-processing of NMR and MS spectra for statistical analysis
  • Statistical analysis of mass spectrometry and NMR data

Quantitative NMR (nuclear magnetic resonance)

Our expertise is to qualify reference substances, determine the purity of molecules and certify their compliance with international quality standards.

Our experts assist pharmaceutical and cosmetic companies in certifying the purity of their reference molecules using NMR methods.

In a nutshell

NMR acquisition parameters

NMR processing parameters

Method validation and parameter verification

Product origin authentification

Optimization of the NMR tool

NMR acquisition parameters

  • Measurement of longitudinal relaxation time
  • Recording of NMR spectra with a very high signal/noise ratio
  • 13C decoupling to avoid the appearance of satellites, etc.
  • Quantitative 1H NMR with very high precision and accuracy (<0.1%)
  • High precision multi-dimensional quantitative NMR

NMR processing parameters

  • Manual phasing of spectra
  • Baseline correction
  • Spectra deconvolution
  • Score correction if 13C satellites are present

Method validation and parameter verification

  • Selectivity
  • Repeatability (precision)
  • Uncertainty calculation

Product origin authentification

  • Non-targeted profiling by high field or benchtop NMR
  • Isotopic analyses on target molecules
  • Quantitative and targeted NMR analysis
  • Development of metabo-isotopomic approaches

Optimization of the NMR tool

  • Ongoing European Research Council project: SUMMIT (Site-specific Ultrasensitive Magnetic resonance of Mixtures for Isotopic Tracking). Development of an analysis workflow based on two NMR methods: dynamic nuclear polarization by dissolution and ultrafast 2D NMR
  • Development of new rapid multi-dimensional methods for targeted and non-targeted metabolomics studies
  • Development of new methods for real-time monitoring of transformations

Publication

Didier G. Diomande, Estelle Martineau, Alexis Gilbert, Pierrick Nun, Ariaki Murata, Keita Yamada, Naoharu Watanabe, Illa Tea, Richard J. Robins, Naohiro Yoshida, and Gérald S. Remaud Position-Specific Isotope Analysis of Xanthines: A 13C Nuclear Magnetic Resonance Method to Determine the 13C Intramolecular Composition at Natural Abundance.v

Analytical Chemistry 2015 87 (13), 6600-6606. DOI: 10.1021/acs.analchem.5b00559

Quantitative 2D NMR (2D nuclear magnetic resonance)

Development of fast and quantitative 2D NMR acquisition methods.

Our experts have developed a protocol for the implementation of targeted and non-targeted metabolic and lipidomic studies.

In a nutshell

Rapid quantification for metabolomics and lipidomics

Quantification of polysaccharides by 2D NMR

  • Concentration measuring of the small molecules in complex samples
  • Exploitation of standard addition methods
  • Development of quantitative approaches for polysaccharide analysis
  • Profiling of biological matrices
  • Targeted and non-targeted analyses

Rapid quantification for metabolomics and lipidomics

  • Production of accurate quantitative data
  • Optimal separation between overlapping resonances
  • Development of fast acquisition methods
  • Representative and complementary experiments: UF COSY, ZF-TOCSY with irregular sampling
  • HSQC with irregular sampling

Quantification of polysaccharides by 2D NMR

  • Characterization of molecular structures
  • Development of powerful quality control methods
  • Measurement of polysaccharide concentration
  • Molecular behavior studies
  • Development of the quantification method
  • Definition of the optimal analytical workflow

Publication

Estelle Martineau, Serge Akoka, Renaud Boisseau, Benoît Delanoue, and Patrick Giraudeau. Fast Quantitative 1H–13C Two-Dimensional NMR with Very High Precision. Analytical Chemistry 2013 85 (9), 4777-4783

Estelle Martineau, Kamel El Khantache, Marion Pupier, Patricia Sepulcri, Serge Akoka, Patrick Giraudeau, Non-linear effects in quantitative 2D NMR of polysaccharides: Pitfalls and how to avoid them, Journal of Pharmaceutical and Biomedical Analysis, Volume 108, 2015, Pages 78-85, ISSN 0731-7085

Estelle Martineau, Jean-Nicolas Dumez, Patrick Giraudeau. Fast quantitative 2D NMR for metabolomics and lipidomics: A tutorial Magnetic Resonance in Chemistry. 04 June 2019

IRMS (Isotope Ratio Mass Spectrometry)

To analyze the 13C, 15N, 2H, 18O and 34S isotopes present in organic matter, for a better understanding of metabolism in the fields of health and nutrition.

The mastery of compound-specific analysis allows to obtain 13C and/or 15N profiles of different matrices.

In a nutshell

Average isotopic composition by EA-IRMS

GC-IRMS analyse

Average isotopic composition by EA-IRMS

  • Sample preparation
  • Analysis 13, 15N, 2H, 18O, 34S

GC-IRMS analyse

  • 15N profiling of amino acids (after hydrolysis of proteins, derivatization, chromatography separation and 15N determination

Publication

Tea, I., Martineau, E., Antheaume, I. et al. 13C and 15N natural isotope abundance reflects breast cancer cell metabolism. Sci Rep 6, 34251 (2016). https://doi.org/10.1038/srep34251

Olivier L Mantha, Caroline Goupille, Jean-François Dumas, Richard Robins, Philippe Bougnoux, Régis Hankard, Arnaud De Luca, Natural isotopic abundances as markers of compliance in clinical trials, The American Journal of Clinical Nutrition, Volume 111, Issue 5, May 2020, Pages 1109–1110

Tea I, Le Guennec A, Frasquet-Darrieux M, Julien M, Romek K, Antheaume I, Hankard R, Robins RJ. Simultaneous determination of natural-abundance δ15N values and quantities of individual amino acids in proteins from milk of lactating women and from infant hair using gas chromatography/isotope ratio mass spectrometry. Rapid Commun Mass Spectrom. 2013 Jun 30;27(12):1345-53.

Engineering at CAPACITÉS

Our Analytical Chemistry experts perform their engineering duties in the very research laboratories of Nantes Université themselves, most notably at CEISAM. They strive to combine technical resources and scientific discovery to meet your R&D needs.

Challenges we’ve met

Quantification of polysaccharides by 2D NMR

Developing a quick quantitative method in two-dimensional NMR
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CAPACITES x Sanofi Pasteur_Quantification des polysaccharides par RMN 2D
All our expertises

Need an innovative and resourceful partner?

Capacités teams inspire, harness and couple numerous cutting-edge skills covering a wide range of applications and expertise to meet all of your innovation needs with passion and commitment!

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    A unique translational expertise in gene therapy

    • Life science

    Our experts in viral vector production work in the gene therapy laboratory in Nantes to support you in the development of new drugs.

    CAPACITÉS’ teams in monitoring host immune response, preclinical assays and viral vector production offer a complete range of support for the development of drug candidates.

    With internationally recognized know-how in the field of gene therapy, our experts offer the responsiveness of a CRO (Contract Research Organization) in the discovery and development phases of gene therapy products.

    Summary

    Services

    Proof of concept

    Preclinical data

    Manufacturing

    Preclinical assays

    Technology platforms

    Viral vectors manufacturing

    In close collaboration with TaRGeT UMR 1089 gene therapy laboratory (UMR INSERM-Nantes Université), we propose the development of processes for the production and characterization of viral gene transfer vectors derived from adenoviruses and AAV. These vectors are intended for applications ranging from basic research to preclinical trials.

    In a nutshell

    AAV vectors

    vector plasmids

    analysis processes

    R&D

    Production of recombinant AAV vectors

    • Testing and proof of concept
    • Testing and proof of concept
    • Hybrid vectors (ITR AAV2)
    • Serotypes 1, 2, 3, 4, 5, 6, 7, 8, 9, rh 10
    • Variants (2i8, DJ, …)
    • Single-stranded and self-complementary rAAV
    • 400 lots per year

    Custom construction of vector plasmids

    • Strategy and design of plasmids
    • Production of plasmids
    • Molecular cloning
    • Amplification de plasmides pour la production de vecteurs

    Development of analysis processes and techniques

    • Development of GMP compatible processes for small/medium scale manufacturing of viral vectors (rAAV)
    • Evaluation of innovative platforms for viral vector amplification (upstream, USP) and purification (downstream, DSP) processes
    • Development of production processes in adherent and suspension cells
    • Development of adapted analytical methods: purity, identity, titre, infectious power
    • Production and characterization of rAAV batches for preclinical efficacy, dose assessment and toxicity studies
    • ICH S6 Directive
    • Biocompatibility studies in final GMP containers

    R&D

    • Research center dedicated to innovation in viral vector production
    • Development of high-performance custom processes for vector production
    • Improved efficiency of rAAV genome wrapping
    • Development of analytical tools based on next generation sequencing and bioinformatics
    • Characterization of genomes and contaminant residues from viral vector manufacturing processes

    Vector production center – CPV

    The research teams and the vector production center of TaRGeT UMR 1089 Gene therapy laboratory work together to ensure reliable, high-quality production and characterization of viral vectors. They can also develop custom analytical tests and new innovative characterization tests.

    In a nutshell

    Upstream technologies

    Downstream technologies

    Upstream technologies

    • HEK293 adherent cells – Cellstacks® vessels (CS) – Up to 24 CS10 per batch for small and medium rAAV production
    • Sf9 cells / BEV platform from 2 to 50 liters per batch for small and medium productions of rAAV
    • HEK293 cells in suspension. Process development phase, cell line screening…
    • 2 benchtop bioreactors: Biostat® B benchtop for 2L/10L scale

    Downstream technologies

    • Clarification: deep filtration, low speed centrifugation, HP homogenization (C55 cell disrupter)
    • Purification: Ion exchange column (IEX), immuno-affinity chromatography (Akta Ready, Akta Pilot, Akta Purifier
    • Akta Explorer), density gradient ultracentrifugation, TFF (KR2i – Spectrum Labs)
    • Formulation, filling in cryovials (multipette/X-Stream®)

    Immune monitoring analyses

    Experts in the monitoring of host immunity in gene therapy protocols, we provide analyses for monitoring immune responses directed against the vector and against the transgene product. We assist in the development of your gene therapy products, from the research project to toxicology studies and clinical trials.

    In a nutshell

    Humoral immunity

    Cellular immunity

    Sample preparation

    Humoral immunity

    • Search for neutralizing factors (AAV, Adenovirus)
    • Detection of anti-AAV and anti-transgen antibodies
    • Cytokines quantification

    Cellular immunity

    • Assessment of the T cell response
    • Multiparametric cell phenotyping

    Sample preparation

    • Isolation of plasma or serum
    • Cell isolation (from whole blood, spleen, lymph nodes, bone marrow)

    Tests available for :

    • Different vectors: AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV8, AAV9, AAV10 and Adenovirus
    • Several types of samples: PBMC (Peripheral Blood Mononuclear Cells), Splenocytes, Lymph node cells, Plasma, Serum, Spinal fluid
    • Adaptable to your needs, in close collaboration with the CPV and TaRGeT UMR 1089 Gene therapy laboratory research teams

    Gene Therapy Immunology Core – GTI

    Analyses for monitoring the immune response against the vector or the transgene product are carried out by the GTI (Gene Therapy Immunology Core) platform of TaRGeT UMR 1089 Gene therapy laboratory. This platform has complete technical means for the Immune monitoring  of gene therapy studies.

    In a nutshell

    ELISpot

    Cell proliferation test

    Tetramer

    flow cytometry

    Cellular neutralization test

    ELISA

    Western Blot

    Luminex

    Plate washer

    Multimode microplate reader

    ELISpot reader

    Turbo gel transfer system

    • ELISpot
    • Cell proliferation test
    • Tetramer
    • Multi-parametric phenotyping by flow cytometry (in collaboration with the Cytocell platform)
    • Cellular neutralization test
    • ELISA
    • Western Blot (WB)
    • Quantification of cytokines by Luminex
    • Plate washer
    • Multimode microplate reader (Spectrophotometry, Fluorescence, Luminescence)
    • ELISpot reader
    • Turbo gel transfer system

    Preclinical analysis

    Evaluation of the biodistribution and expression profile of transgenes is necessary to understand the pharmacology and pharmacokinetics of a therapeutic product. We are recognized as experts in the molecular and biochemical analyses required to assess the efficacy and biosafety of gene therapy products.

    In a nutshell

    Evaluation of functionality

    Evaluation of biodistribution

    Expression Profile Assessment

    development of preclinical studies

    Absolute quantification

    • Evaluation of the functionality of therapeutic products
    • Evaluation of biodistribution and dissemination of gene therapy products after administration
    • Expression Profile Assessment:
    • RT-qPCR (relative quantification of the transgenic messenger)
    • Western-Blot or ELISA on tissues or fluids (specific detection of the transgenic protein)
    • Absolute quantification of the transgenic genome: qPCR on tissues and biological fluids
    • Global coordination and expertise for the development of preclinical studies to evaluate a therapeutic product

    Preclinical analysis core – PAC

    Our leading expertise, in the evaluation of biodistribution and expression profiles of transgenes, is implemented using multiple tools and equipment available on PAC (Preclinical Analytics Core) platform of the Nantes UMR1089 gene therapy laboratory.

    In a nutshell

    Tissue shredder

    Pipetting machine

    Nanophotometer

    microplate reader

    Colorimetric and fluorimetric reader

    Electrochemiluminescence reader

    PCR and qPCR instruments

    Turbo gel transfer system

    • Tissue shredder (Qiagen, Tissue Lyser II)
    • Pipetting machine (Hamilton, Microlab Star)
    • Nanophotometer and microplate reader (Thermo Scientific, Multiskan FC)
    • Colorimetric and fluorimetric reader (Tecan, Spark microplate reader)
    • Electrochemiluminescence reader (MSD, quickplex reader)
    • PCR and qPCR instruments (96 and 384 wells, Applied Biosystems, Biorad)
    • Turbo gel transfer system (Trans-blot turbo system, Biorad)

    Engineering at CAPACITÉS

    Our gene therapy experts carry out their engineering work directly in the research laboratories of Nantes Université, particularly in TaRGeT UMR 1089 Gene therapy laboratory
    On a daily basis, they combine technical resources and scientific discoveries to meet your R&D challenges.

    Need an innovative and resourceful partner?

    Capacités teams inspire, harness and couple numerous cutting-edge skills covering a wide range of applications and expertise to meet all of your innovation needs with passion and commitment!

    or send us a message:






      Required fields