Attenuated Total Reflection Fourier Transform Infrared (ATR-FTIR) spectroscopy is a label-free, non-destructive technique that can be applied to a vast range of biological applications, from imaging cancer tissues and live cells, to determining protein content and protein secondary structure composition. This review summarises the recent advances in applications of ATR-FTIR spectroscopy to biopharmaceuticals, the application of this technique to biosimilars, and the current uses of FTIR spectroscopy in biopharmaceutical production. We discuss the use of ATR-FTIR spectroscopic imaging to investigate biopharmaceuticals, and finally, give an outlook on the possible future developments and applications of ATR-FTIR spectroscopy and spectroscopic imaging to this field. Throughout the review comparisons will be made between FTIR spectroscopy and alternative analytical techniques, and areas will be identified where FTIR spectroscopy could perhaps offer a better alternative in future studies. This review focuses on the most recent advances in the field of using ATR-FTIR spectroscopy and spectroscopic imaging to characterise and evaluate biopharmaceuticals, both in industrial and academic research based environments.
Monoclonal
antibodies (mAbs) have been used as therapeutics for
the last few decades. It is necessary to investigate the stability
of these mAbs under stress conditions and to elucidate aggregation
mechanisms as a means of developing approaches which minimize the
problem. Attenuated total reflection (ATR)-FTIR spectroscopic imaging
allows probing of a sample at a depth of penetration of around 0.5–5
μm, which makes it suitable for the study of aggregated proteins
when accumulated as a layer close to the surface of the ZnSe internal
reflection element (IRE). Here, macro ATR-FTIR spectroscopic imaging,
along with a variable angle of incidence accessory, have been used
to differentiate between the secondary structure of proteins in bulk
solution and those that have precipitated onto or near the ZnSe IRE
surface. IgG spectra obtained from protein samples in individual wells
have been averaged, extracted, and preprocessed, and the Amide I bands
of the protein samples were compared and further analyzed to reveal
protein distribution at the ZnSe IRE surface. These findings show
depth profiling of IgG aggregates at the ZnSe IRE surface (0.5–5
μm) and do not follow a trend of decreasing protein presence
with an increasing angle of incidence or increasing depth of penetration,
suggesting an irregular distribution of aggregates in the z-direction.
Biopharmaceuticals are used to treat a range of diseases from arthritis to cancer, however, since the advent of these highly specific, effective drugs, there have been challenges involved in their...
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