Extractable and Leachable

Invitron Advanced Analytics specializes in complex characterization of peptides and protein. Our characterization services include techniques to characterize or compare peptide/protein higher order structure (HOS). These include methods to characterize peptide/protein folding (secondary/tertiary structure), peptide/protein stability, peptide/protein conformation, peptide/protein aggregation, and the oligomeric state of the native peptide/protein (quaternary structure).

Services

Primary Peptide Structure

Peptide mapping is a powerful technique that can be used to comprehensively identify the primary structure (amino acid sequence) of a peptide/protein. Workflows employ bottom-up methodologies including enzymatic digestion followed by separation of the resulting peptides and analysis via ultraviolet (UV) detection and/or mass spectrometry (MS). 

Higher Order Structure

Secondary Structure
  • Far-UV CD (190 – 250 nm) (Far-Ultraviolet Circular Dichroism)
  • F T I R (Fourier Transform Infrared Spectroscopy)
  • Raman Spectroscopy
Tertiary Structure
  • Near-UV CD (250 – 350 nm) (Near-Ultraviolet Circular Dichroism)
  • Intrinsic Fluorescence
  • N M R (Nuclear Magnetic Resonance Spectroscopy)
    • C D Services
    • Circular dichroism (CD) spectroscopy for characterizing the higher order structure (HOS) and thermal stability of peptide/proteins and peptide.

    Far-UV CD measurements are obtained using circularly polarized light over a wavelength range of about ≤ 200-250 nm. Light absorption in this wavelength range is mainly due to absorption by the peptide bonds. Since each type of secondary structure element (such as α-helix, β-sheet, etc.) has a distinctive spectral CD profile, Far-UV CD is sensitive to changes in the secondary structure.

    Near-UV CD measurements are obtained over a wavelength range of about 250-300 nm, where tyrosine, tryptophan and phenylalanine residues, as well as disulphide bridges absorb light and are CD sensitive.

    Aggregation Analysis

    Invitron Advanced Analytics has extensive experience in analysis of aggregation in peptide/proteins and other macromolecules. The amount, type, and size of the aggregates can have important consequences for the safety and efficacy of the molecule.

    Light Scattering

    • SEC-UV/MALS (Size Exclusion Chromatography-Ultraviolet/Multi-angle Light Scattering)
    • CG-MALS (Composition-Gradient MALS)
    • DLS (Dynamic Light Scattering)

      Analytical Ultracentrifugation

      • SE-AUC (Sedimentation Equilibrium-Analytical Ultracentrifugation)
      • SV-AUC (Sedimentation Velocity-Analytical Ultracentrifugation) Gel Electrophoresis
      • Denaturing – SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis)
      • Non-Denaturing or Native
          -FFF (Field Flow Fractionation)
      • AF4 (Asymmetric Field Flow Fractionation)
          -MFI (Micro-Flow Imaging)

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