CONFOCHECK Infrared Protein Analysis Innovation with Integrity

CONFOCHECK: FTIR System for Protein Analytics FTIR Protein Analysis Infrared spectroscopy measures molecular vibrations caused by the specific absorption of infrared (IR) radiation by chemical bonds. For over 35 years, it has been known that the shape and frequency of the amide I band, which is assigned to the C=O stretching vibration within peptide bonds, is characteristic for the structure of the studied protein. From this single band the presence and relative percentage of secondary structure elements (α-helix, β-strand) in a protein can be determined and structural changes detected with high...

FTIR Protein Analysis Information Content of IR Protein Spectra The information content of the IR protein spectrum is not limited to the amide I band. Since IR spectroscopic data follows Lambert-Beer´s law, the protein spectrum can be utilized for concentration determination. Also, side chains of various amino acids can be monitored and directly compared under varying environmental conditions (pH, salts, ligands), or in mutant proteins with altered primary sequence. Moreover, all other chemical components used in protein samples like certain pH buffers or proteinstabilizing agents are also represented...

FTIR Protein Determination and Structure Analysis The demanding research lab and biopharmaceutical environments necessitate that an IR system provides accurate and reproducible protein measurements, rapid analysis (< 1 minute), and ease of use. The CONFOCHECK system satisfies these demands. Unique in that it combines the necessary hardware with software for precise spectral measurement and interpretation using internal calibration models. The stability of the system permits high sensitivity (dynamic range: 0.03 to >200 µg/µl) and direct analysis of proteins in any biological buffer with a spectrum...

Temperature Ramp Fig. 3A: Antibody spectra in temperature ramp Thermal stability studies of a therapeutic antibody using the BioATR II accessory. The CONFOCHECK software permits the user designated configuration of temperature ramps. When the temperature range (maximum from 0° - 95° C), increment and equilibration time are set, the system starts to acquire automatically a spectrum at each predefined temperature. The IR spectra of an antibody at different temperatures (25-95° C) are shown (Figure 3A). As indicated by the unchanged amide I band shape the protein maintains its native-sheet structure...

CONFOCHECK: A Dedicated FTIR System for Protein Analysis The CONFOCHECK is a compact IR spectrometer system dedicated to the analysis of proteins in aqueous solutions. Its robust design permits fast data acquisition while maintaining high sensitivity thus making high sample throughput possible. The system is controlled by a user friendly software interface that guides the user through the measurement procedure for spectral acquisition and temperature ramps. The system includes internal calibrations for protein concentration determination and secondary structure prediction, greatly simplifying...

CONFOCHECK utilizes three FTIR sampling accessory cells to cover all protein applications. ■ Protein quantification (down to 0.03 mg/ml in aqueous solution) ■ Detection of conformational changes in proteins AquaSpec cell This flow through transmission cell is used to study proteins in aqueous solution. The analysis of conformational stability, concentration and secondary structure composition are typical applications. For biocompatibility, all tubes are made from PTFE. In-line filters are integrated to prevent contamination of the cell volume by aggregated and precipitated protein. This ATR-accessory...

Application Support Bruker is staffed by expert life scientists and engineers that have in depth knowledge of protein analytics and instrumentation. Our product specialists are prepared to assist you with method development, selection and usage of sampling accessories, choices of optical components and software operation. We offer customized instruction and support packages to fit your needs. Bruker systems are designed to provide many years of trouble-free operation, but should a problem occur a network of Bruker Optics offices and representatives throughout the world are ready to promptly respond...