Proteomics

In this application note, we highlight the high sensitivity of the Duetta for measuring transferrin at very low concentrations. The Duetta spectrofluorometer combines both fluorescence and absorbance measurements simultaneously, thus providing key advantages for protein analysis. In parallel, we compare the fluorescence results obtained using the Duetta with absorbance results obtained with a dedicated low-volume, benchtop spectrophotometer.

Duetta is an analytical instrument capable of recording both the fluorescence and the absorbance of a sample. Absorbance data provides concentration-dependant information on non-fluorescent compounds. Through its user-friendly software, Duetta allows different measurements related to biomolecules like DNA, RNA and proteins including protein concentration, DNA/RNA purity, finding the concentration of an unknown sample based on a standard-based calibration curve, Pass/Fail testing and all standard fluorescence and absorbance related measurements. All these functionalities are possible through dedicated applications in EzSpec™ software for Duetta.

The prevalence of allergies in the world is between 30 and 40%. Even though it exists medical treatments such as antihistaminic and desensitization this number is constantly increasing. Thus, in this application note, the identification and chemical characterization of aeroallergens by Raman microscopy will allow to prevent people who are affected by respiratory allergies of the presence of this type of allergens in indoor and outdoor air.

The new HORIBA Duetta fluorescence and absorbance spectrometer offers many unique benefits for molecular spectroscopy. It is primarily thought of as a spectrofluorometer that combines absorbance and fluorescence spectroscopy to correct the fluorescence fingerprint for concentration-related effects. Duetta can also be used as a precise absorbance spectrometer. This application note explores the use of Duetta for a common absorbance application, the Protein A280 application.

The use of low molecular weight molecules in the industries of drug discovery and agri-food has become increasingly important over the course of a century of drug research and science development in agriculture and in pharmaceutical field.

The majority of proteins are glycosylated: they possess oligosaccharide chains and are hence termed glycoproteins. Oligosaccharides, also called carbohydrate or sugar, are often quite large (as large as some protein domains for example) and they have many functions in molecular interactions.

Description of the advantages of combining the use of peptide chips with direct label-free detection as achieved by SPR imaging (SPRi).

This application note shows that the HORIBA Scientific SPRi platform is suitable for the analysis of aptamer-based molecular interactions. For this purpose, the interaction between human IgE protein, an antibody involved in allergic reactions, and a human IgE specific aptamer4 is presented as proof-of-concept.

Our SPRi technology (Surface Plasmon Resonance imaging), is able to measure multiplexed label-free biolo-gical interactions in a complex media such as serum.We will demonstrate the ability to monitor specific interactions between immobilized peptides and proteins issued from non purified immune sera, using a cystamine/glutaraldehyde SAM surface chemistry on the bio-chip surface. We chose to illustrate this ability by monitoring the interaction between an ovalbumin peptide fragment and an ovalbumin immunized serum.

The purpose of this proof of concept is to show that Surface Plasmon Resonance imaging (SPRi) is a suitable technique to detect allergens. Compared with other techniques, such as ELISA or Luminex, which are based on colorimetric or fluorescence detection, this technology does not require the labeling of antibodies.

HORIBA provides the pharmaceutical industry, biotech companies and academic research laboratories with high performance instrumentation based on SPRi technology (Surface Plasmon Resonance imaging). To demonstrate the power of this technology, we carried out an experiment based on an antibody-antigen interaction. Original surface chemistry combined with an electrochemical process allows the rapid coupling of biomolecules to the gold layer (polypyrrole co-polymerisation) on the top of a glass prism.

Rapid mixing accessories to perform stopped flow measurements have found application in characterizing interactions and reactions occurring in solution. Reactants are expelled from syringes, mixed and injected into a flowcell.

The potential health benefits stemming from the antioxidant activity of curcumin, commonly found in turmeric (Curcuma longa L), has attracted the interest of several research groups.

Polarized light striking a fluorescent molecule results in polarized fluorescence. This polarized emission gradually returns to unpolarized fluorescence depending on rotational diffusion and other factors. Anisotropy is directly related to the polarization, and is the ratio of the polarized light component to the total light intensity.