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CONFOCHECK for Protein Analysis

Analyze Protein in Water with CONFOCHECK

The CONFOCHECK is a dedicated FT-IR system for the investigation of proteins in water. Its specific configuration facilitates a fast data acquisition (ca. 30 sec. per sample) with a high sample throughput controlled by an extremely user-friendly software interface.

About CONFOCHECK and Proteins

Main applications for the CONFOCHECK are the detection and quantification of conformational changes in proteins and the protein concentration determination. Quantitative determination of the protein concentration and the secondary structure elements (alpha-helix, beta-sheet) from the analyzed protein samples is accomplished within seconds via internal calibrations.

The CONFOCHECK is equipped with several specific protein sampling cuvettes that allow the measurement of solved and membrane bound proteins, even at very low amounts (10 µg). To reach optimal temperature constancy and to facilitate the measurement of temperature induced protein conformational changes, the sample cuvettes are temperature-controlled by a thermostat.

FT-IR Protein Applications

  • Protein quantification
  • Detection of conformational changes
  • Protein dynamics (temperature induced conformational changes)
  • Monitoring of conformational changes during protein aggregation, precipitation and crystallization
  • Determination of the secondary structure
  • Quantification of all kind of solutes in aqueous samples (buffers, excipients, detergents)

Analyzing Proteins in Pharma

  • Pre-formulation and formulation development: Analysis of the influence of the formulation on protein stability
  • Stability studies (stress tests): Influence of pH, temperature, mutation, etc. on the protein stability
  • Quality control: Check of protein concentration, structure on correct product composition (buffer, excipients) in production environment
  • Binding studies: Analysis of the impact of ligand-binding on the protein structure
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Protein Analysis in Water: AquaSpec™

The AquaSpecTM is used for the measurement of water soluble proteins or solubilized membrane-bound proteins. Using the internal calibrations of the CONFOCHECK the secondary structure and concentration of the analyzed protein are determined directly after measurement (ca. 30 sec).

It features a flow-through transmission cell, with an optimal path length of ~7 µm. The AquaSpecTM also permits higher sample throughput and accuracy. Easily measure low protein concentrations (below 0.1 mg/ml) and detect the smallest conformational changes.

Folding in Proteins: BioATRCell II

Temperature-induced unfolding / refolding processes are studied by the BioATRCell II. Apply automated temperature ramps (0-95°C) to follow specific changes of the secondary structure and determine the proteins melting point.

Furthermore, you can characterize fibrillation and aggregation processes and monitor protein structural changes in dependence of pH‑changes or ligand-binding interactions. Using a special dialysis top section for BioATRCell II, low molecular weight substances can be dialyzed into the protein compartment without diluting the protein.

More information

About CONFOCHECK.

About the INVENIO FT-IR platform.

AquaSpec Transmission Cell

The AquaSpecTM micro transmission cell is a flow-through cuvette for the measurement of aqueous protein sample.

Using the AquaSpecTM cell IR-spectroscopic measurements of water soluble proteins, solubilised membrane proteins and proteins immobilised on nano particles can be performed. It has a path length of approx. 7 µm that is optimized for the measurement of aqueous solutions. For the measurement the sample (ca. 10 µg protein) is filled into the cell via an injection port. The cell is cleaned afterwards by injection of the buffer solution. All parts that get into contact with the sample are made from bio compatible materials to avoid unspecific interactions with the protein. The AquaSpecTM cell combines a minimized sample volume, high pressure stability (> 30 bar) and an extreme path length precision. The windows of the cell are made of CaF2.

Applications

  • Determination of changes in protein structure
  • Protein concentration determination
  • Quality control of biopharmaceuticals
  • Secondary structure determination
  • Quantification of various substances in aqueous solutions

The AquaSpecTM cell is integral part of CONFOCHECK, a FTIR system that is specially designed for protein analysis. The CONFOCHECK is based on a high performance FTIR spectrometer that allows very fast acquisition (measurement time 30 sec) of protein spectra with excellent S/N (Signal/Noise). It is tightly sealed and includes a precise water bath that allows temperature control of the AquaSpecTM transmission cell therefore guaranteeing optimal measurement stability.

Using the AquaSpecTM transmission cell the highest sensitivity for the detection of conformational changes and the best accuracy in the protein concentration determination is achieved.

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BioATRCell II

The BioATRCell II allows to follow temperature-induced conformational changes of solved and membrane-standing proteins between 0 - 95°C.

The BioATRCell II is a micro-ATR unit that was specially developed for the investigation of temperature-induced unfolding, refolding and denaturation processes of proteins in aqueous media. It can be applied for water solved and also for membrane-standing proteins. The number of internal reflections in the ATR crystal (silicon) is adjusted to result in an optimal protein signal in aqueous samples. The BioATRCell II is temperature-controlled by the same thermostat like the other accessories that are part of the CONFOCHECK system. With this unit temperature ramps from 0 to 95°C can be performed automatically to analyze the temperature-induced structural changes and the protein melting point.

Applications:

  • Automated temperature ramps (0-95°C):
    Analysis of temperature-induced conformational changes
    Measurement of phase transitions in lipids
  • Quality control of biopharmaceuticals
  • Membrane-standing proteins
  • Conformational changes induced by ligand-binding, pH-changes, salts

The BioATRCell II is a micro-ATR unit that was specially developed for the investigation of temperature-induced unfolding, refolding and denaturation processes of proteins in aqueous media. It can be applied for water solved and also for membrane-standing proteins. The number of internal reflections in the ATR crystal (silicon) is adjusted to result in an optimal protein signal in aqueous samples. The BioATRCell II is temperature-controlled by the same thermostat like the other accessories that are part of the CONFOCHECK system. With this unit temperature ramps from 0 to 95°C can be performed automatically to analyze the temperature-induced structural changes and the protein melting point.

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