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Quantum Cascade Laser Based Infrared Spectroscopy: A New Paradigm for Protein Secondary Structure Measurement

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Abstract

Mid-infrared spectroscopy is one of the major analytical techniques employed for measurements of protein structure in solution. Traditional Fourier Transform-Infrared (FT-IR) measurement is limited by its blackbody light source that is inherently spatially incoherent and has low optical power output. This limitation is pronounced when working with proteins in aqueous solutions. Strong absorbance of water in protein amide I region 1600–1700 cm−1 restricts light path length to <10 μm and imposes significant experimental challenges in sample and flow cell handling. Emerging laser spectroscopic techniques use high-power coherent laser as light source that overcomes the limitation in FT-IR measurement. In this study, we employed an innovative infrared spectrometer that uses quantum cascade laser (QCL) as light source. Continuous infrared radiation from this laser source can be swiftly swept within the amide I region (1600–1700 cm−1) and amide II region (1500–1600 cm−1), which makes this technique ideal for protein secondary structure study. Protein solutions as low as 0.5 mg/mL were measured rapidly without any sample preparation. Infrared spectra of model proteins were thus collected, and a chemometric model based on partial least squares regression was developed to quantify α-helix and β-strand motifs in protein secondary structure. The model was applied to measurement of the native secondary structure of commercial therapeutic proteins and bovine serum albumin (BSA) and in thermal degradation studies.

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Funding

This work is the result of research projects funded by Bristol-Myers Squibb Company.

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Authors and Affiliations

  1. Global Quality Analytical Science & Technology, Bristol Myers Squibb, New Brunswick, New Jersey, 08901, USA

    Chunguang Jin, Amrish Patel, Jeremy Peters & Ravi Kalyanaraman

  2. DRS Daylight Solutions, San Diego, California, 92127, USA

    Santosh Hodawadekar

Authors
  1. Chunguang Jin
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  2. Amrish Patel
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  3. Jeremy Peters
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  4. Santosh Hodawadekar
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  5. Ravi Kalyanaraman
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The manuscript was written through contributions of all authors. All authors have given approval to the final version of the manuscript.

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Correspondence to Chunguang Jin or Ravi Kalyanaraman.

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Jin, C., Patel, A., Peters, J. et al. Quantum Cascade Laser Based Infrared Spectroscopy: A New Paradigm for Protein Secondary Structure Measurement. Pharm Res 40, 1507–1517 (2023). https://doi.org/10.1007/s11095-022-03422-8

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