7− 9įourier transform infrared (FT-IR) spectroscopy, the most Of the amide I and II bands, particularly with chemometric analysis. However, it has been shown that additionalĪnd more comprehensive information can be gained by combined analysis The individual secondary structure components, curve fitting 4 or chemometric approaches such as partial least-squares 5 or multivariate curve resolutions-alternating Orientations in the α-helices, β-sheets, turns, and random Of hydrogen bonding, dipole–dipole interactions, and geometric To secondary structure due to changes in the interaction of the C=OĪnd N–H groups of the amide bond induced by different arrangements 2 The position and shape of the amide bands show sensitivity Recognized to be sensitive to secondary structure. The amide II band (1600–1500 cm –1) has been The most prominentīand for secondary structure analysis is the amide I band (1700–1600 The IR spectrum that are referred to as amide bands. Polypeptides and proteins show characteristic absorption bands in (4000–400 cm –1) provides detailed informationĪbout the specific rotational–vibrational transitions of molecules. Quantitative analyses of proteins in complex matrices such as thoseįound in up- and downstream bioprocess monitoring and similar challengingĪpplications which can not be readily met by conventional FT-IR spectroscopy. Protein–ligand interaction studies as well as qualitative and This opens a wide range of future applications including Previous research type EC-QCL setups as well as commercially available Large optical path lengths, and short measurement times and thus outperforms The presented setup combines high sensitivity, Of the protein measurements and to ensure maximum flexibility forįuture applications. Time of the setup can be adapted to fulfill the required sensitivity Furthermore, a highly linear response was demonstrated for concentrationsīetween 0.05 and 10 mg mL –1. Identified at concentrations as low as 0.1 mg mL –1. Of proteins with different secondary structures were successfully Way, noise levels better by a factor of up to 20 were achieved compared High-intensity laser light source, a thermoelectrically cooled mercuryĬadmium telluride balanced detection module was employed. For minimizing the noise introduced by the Optical setup that adjusts the path length (26 μm) to ensure Laser (EC-QCL) operated at room temperature with an optimized double-beam This large improvement is made possibleīy combining the latest-generation external cavity-quantum cascade Our previous results and other state-of-the-art mid-IR-based techniquesīy almost an order of magnitude). Of detection as low as 0.0025 mg mL –1 (outperforming Report a mid-IR transmission setup for the analysis of the proteinĪmide I and amide II band in aqueous solutions that achieves a limit
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