Abstract
Changes in protein concentrations within human blood are used as an indicator for nutritional state, hydration and underlying illnesses. They are often measured at regular clinical appointments and the current analytical process can result in long waiting times for results and the need for return patient visits. Attenuated total reflectance – Fourier transform infrared (ATR-FTIR) spectroscopy has the ability to detect minor molecular differences, qualitatively and quantitatively, in biofluid samples, without extensive sample preparation. ATR-FTIR can return an analytical measurement almost instantaneously and therefore could be deemed as an ideal technique for monitoring molecular alterations in blood within the clinic. To determine the suitability of using ATR-FTIR spectroscopy to enable protein quantification in a clinical setting, pooled human serum samples spiked with varying concentrations of human serum albumin (HSA) and immunoglobulin G (IgG) were analysed, before analysing patient clinical samples. Using a validated partial least squares method, the spiked samples (IgG) produced a linearity as high as 0.998 and a RMSEV of 0.49 ± 0.05 mg mL−1, with the patient samples producing R2 values of 0.992 and a corresponding RMSEV of 0.66 ± 0.05 mg mL−1. This claim was validated using two blind testing models, leave one patient out cross validation and k-fold cross validation, achieving optimum linearity and RMSEV values of 0.934 and 1.99 ± 0.79 mg mL−1, respectively. This demonstrates that ATR-FTIR is able to quantify protein within clinically relevant complex matrices and concentrations, such as serum samples, rapidly and with simple sample preparation. The ability to provide a quantification step, along with rapid disease classification, from a spectroscopic signature will aid clinical translation of vibrational spectroscopy to assist with problems currently faced with patient diagnostic pathways.
| Original language | English |
|---|---|
| Pages (from-to) | 50-58 |
| Number of pages | 9 |
| Journal | Vibrational Spectroscopy |
| Volume | 99 |
| Early online date | 1 Sep 2018 |
| DOIs | |
| Publication status | Published - 30 Nov 2018 |
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Keywords
- attenuated total reflection
- clinical
- infrared
- protein
- quantification
- serum
Cite this
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Enabling quantification of protein concentration in human serum biopsies using attenuated total reflectance – Fourier transform infrared (ATR-FTIR) spectroscopy. / Spalding, Katie; Bonnier, Franck; Bruno, Clément; Blasco, Hélène; Board, Ruth; Benz-de Bretagne, Isabelle; Byrne, Hugh J.; Butler, Holly J.; Chourpa, Igor; Radhakrishnan, Pretheepan; Baker, Matthew J.
In: Vibrational Spectroscopy, Vol. 99, 30.11.2018, p. 50-58.Research output: Contribution to journal › Article
TY - JOUR
T1 - Enabling quantification of protein concentration in human serum biopsies using attenuated total reflectance – Fourier transform infrared (ATR-FTIR) spectroscopy
AU - Spalding, Katie
AU - Bonnier, Franck
AU - Bruno, Clément
AU - Blasco, Hélène
AU - Board, Ruth
AU - Benz-de Bretagne, Isabelle
AU - Byrne, Hugh J.
AU - Butler, Holly J.
AU - Chourpa, Igor
AU - Radhakrishnan, Pretheepan
AU - Baker, Matthew J.
PY - 2018/11/30
Y1 - 2018/11/30
N2 - Changes in protein concentrations within human blood are used as an indicator for nutritional state, hydration and underlying illnesses. They are often measured at regular clinical appointments and the current analytical process can result in long waiting times for results and the need for return patient visits. Attenuated total reflectance – Fourier transform infrared (ATR-FTIR) spectroscopy has the ability to detect minor molecular differences, qualitatively and quantitatively, in biofluid samples, without extensive sample preparation. ATR-FTIR can return an analytical measurement almost instantaneously and therefore could be deemed as an ideal technique for monitoring molecular alterations in blood within the clinic. To determine the suitability of using ATR-FTIR spectroscopy to enable protein quantification in a clinical setting, pooled human serum samples spiked with varying concentrations of human serum albumin (HSA) and immunoglobulin G (IgG) were analysed, before analysing patient clinical samples. Using a validated partial least squares method, the spiked samples (IgG) produced a linearity as high as 0.998 and a RMSEV of 0.49 ± 0.05 mg mL−1, with the patient samples producing R2 values of 0.992 and a corresponding RMSEV of 0.66 ± 0.05 mg mL−1. This claim was validated using two blind testing models, leave one patient out cross validation and k-fold cross validation, achieving optimum linearity and RMSEV values of 0.934 and 1.99 ± 0.79 mg mL−1, respectively. This demonstrates that ATR-FTIR is able to quantify protein within clinically relevant complex matrices and concentrations, such as serum samples, rapidly and with simple sample preparation. The ability to provide a quantification step, along with rapid disease classification, from a spectroscopic signature will aid clinical translation of vibrational spectroscopy to assist with problems currently faced with patient diagnostic pathways.
AB - Changes in protein concentrations within human blood are used as an indicator for nutritional state, hydration and underlying illnesses. They are often measured at regular clinical appointments and the current analytical process can result in long waiting times for results and the need for return patient visits. Attenuated total reflectance – Fourier transform infrared (ATR-FTIR) spectroscopy has the ability to detect minor molecular differences, qualitatively and quantitatively, in biofluid samples, without extensive sample preparation. ATR-FTIR can return an analytical measurement almost instantaneously and therefore could be deemed as an ideal technique for monitoring molecular alterations in blood within the clinic. To determine the suitability of using ATR-FTIR spectroscopy to enable protein quantification in a clinical setting, pooled human serum samples spiked with varying concentrations of human serum albumin (HSA) and immunoglobulin G (IgG) were analysed, before analysing patient clinical samples. Using a validated partial least squares method, the spiked samples (IgG) produced a linearity as high as 0.998 and a RMSEV of 0.49 ± 0.05 mg mL−1, with the patient samples producing R2 values of 0.992 and a corresponding RMSEV of 0.66 ± 0.05 mg mL−1. This claim was validated using two blind testing models, leave one patient out cross validation and k-fold cross validation, achieving optimum linearity and RMSEV values of 0.934 and 1.99 ± 0.79 mg mL−1, respectively. This demonstrates that ATR-FTIR is able to quantify protein within clinically relevant complex matrices and concentrations, such as serum samples, rapidly and with simple sample preparation. The ability to provide a quantification step, along with rapid disease classification, from a spectroscopic signature will aid clinical translation of vibrational spectroscopy to assist with problems currently faced with patient diagnostic pathways.
KW - attenuated total reflection
KW - clinical
KW - infrared
KW - protein
KW - quantification
KW - serum
UR - http://www.scopus.com/inward/record.url?scp=85052879660&partnerID=8YFLogxK
U2 - 10.1016/j.vibspec.2018.08.019
DO - 10.1016/j.vibspec.2018.08.019
M3 - Article
VL - 99
SP - 50
EP - 58
JO - Vibrational Spectroscopy
JF - Vibrational Spectroscopy
SN - 0924-2031
ER -