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Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy: An analytical technique to understand therapeutic responses at the molecular level.

Kalmodia S, Parameswaran S, Yang W, Barrow CJ, Krishnakumar S - Sci Rep (2015)

Bottom Line: Fourier Transform Infrared (FTIR) spectroscopy has become a popular technique in the field of cancer therapy with an ability to elucidate molecular interactions.Towards achieving the aim, we utilized the mouse xenograft model of retinoblastoma and nanoparticle mediated targeted therapy.The results indicate that the mechanism underlying the response differed between the treated and untreated group which can be elucidated by unique spectral signatures generated by each group.

View Article: PubMed Central - PubMed

Affiliation: Department of Nano biotechnology, Vision Research Foundation, Sankara Nethralaya, 18, College Road, Nungambakkam, Chennai - 600 006, India.

ABSTRACT
Rapid monitoring of the response to treatment in cancer patients is essential to predict the outcome of the therapeutic regimen early in the course of the treatment. The conventional methods are laborious, time-consuming, subjective and lack the ability to study different biomolecules and their interactions, simultaneously. Since; mechanisms of cancer and its response to therapy is dependent on molecular interactions and not on single biomolecules, an assay capable of studying molecular interactions as a whole, is preferred. Fourier Transform Infrared (FTIR) spectroscopy has become a popular technique in the field of cancer therapy with an ability to elucidate molecular interactions. The aim of this study, was to explore the utility of the FTIR technique along with multivariate analysis to understand whether the method has the resolution to identify the differences in the mechanism of therapeutic response. Towards achieving the aim, we utilized the mouse xenograft model of retinoblastoma and nanoparticle mediated targeted therapy. The results indicate that the mechanism underlying the response differed between the treated and untreated group which can be elucidated by unique spectral signatures generated by each group. The study establishes the efficiency of non-invasive, label-free and rapid FTIR method in assessing the interactions of nanoparticles with cellular macromolecules towards monitoring the response to cancer therapeutics.

No MeSH data available.


Related in: MedlinePlus

Principal component analysis (PCA) on the raw spectral data before 2nd derivatization.The raw spectral data (wavelength range 3040-930 cm−1) (A) was pre-processed to obtain the “bioband” spectra (wavelength: 1800-930 cm−1; 3040-2810 cm−1) corresponding to the major biomolecules such as proteins, lipids and nucleic acids (B). The spectra corresponding to bioband range was further analyzed by extended multiplicative scatter correction (EMSC) to obtain only the spectra from chemical information (C). PCA analysis of the EMSC corrected spectra revealed maximum difference in the PCs between the Control, GNPs-1 and GNPs-2 as revealed by the Cumulative variance plot (D) and PCA score plot (E).
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f4: Principal component analysis (PCA) on the raw spectral data before 2nd derivatization.The raw spectral data (wavelength range 3040-930 cm−1) (A) was pre-processed to obtain the “bioband” spectra (wavelength: 1800-930 cm−1; 3040-2810 cm−1) corresponding to the major biomolecules such as proteins, lipids and nucleic acids (B). The spectra corresponding to bioband range was further analyzed by extended multiplicative scatter correction (EMSC) to obtain only the spectra from chemical information (C). PCA analysis of the EMSC corrected spectra revealed maximum difference in the PCs between the Control, GNPs-1 and GNPs-2 as revealed by the Cumulative variance plot (D) and PCA score plot (E).

Mentions: A total of 1493 raw spectra were obtained between wavelengths (3040-930 cm−1) from all three groups (Fig. 4A). This included 543 from vehicle control, 486 from GNPs-1 and 464 from GNPs-2. This data was further analyzed in the bioband range (1800-930 and 3040-2810 cm−1) (Fig. 4B) followed by the EMSC correction (Fig. 4C) and PCA analysis on the raw data (Fig. 4D,E). PCA was performed on the spectral data of “BioBands” through cross validation at 7 PCs. The interval of the wavelength was kept at 1–60 and 320–548 in the ranges of wave numbers: 3040-2810 cm−1 and 1800-930 cm−1, respectively. The actual bioband wave number was 3039.234 - 2811.677 and 1808.883 - 929.51 cm−1. The specific range was selected since, it is indicated in the identification of lipids, nucleic acid and protein molecules present in the mammalian cell2728.PCA was applied to all the data and provided 3 levels with 95.22% of variance, best observed in three-dimensional space (PC1 versus PC2 versus PC3) (Fig. 4D,E). The PC score plot of unpriced original data showed clear scattering at different PC levels for all the three groups studied.


Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy: An analytical technique to understand therapeutic responses at the molecular level.

Kalmodia S, Parameswaran S, Yang W, Barrow CJ, Krishnakumar S - Sci Rep (2015)

Principal component analysis (PCA) on the raw spectral data before 2nd derivatization.The raw spectral data (wavelength range 3040-930 cm−1) (A) was pre-processed to obtain the “bioband” spectra (wavelength: 1800-930 cm−1; 3040-2810 cm−1) corresponding to the major biomolecules such as proteins, lipids and nucleic acids (B). The spectra corresponding to bioband range was further analyzed by extended multiplicative scatter correction (EMSC) to obtain only the spectra from chemical information (C). PCA analysis of the EMSC corrected spectra revealed maximum difference in the PCs between the Control, GNPs-1 and GNPs-2 as revealed by the Cumulative variance plot (D) and PCA score plot (E).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4645174&req=5

f4: Principal component analysis (PCA) on the raw spectral data before 2nd derivatization.The raw spectral data (wavelength range 3040-930 cm−1) (A) was pre-processed to obtain the “bioband” spectra (wavelength: 1800-930 cm−1; 3040-2810 cm−1) corresponding to the major biomolecules such as proteins, lipids and nucleic acids (B). The spectra corresponding to bioband range was further analyzed by extended multiplicative scatter correction (EMSC) to obtain only the spectra from chemical information (C). PCA analysis of the EMSC corrected spectra revealed maximum difference in the PCs between the Control, GNPs-1 and GNPs-2 as revealed by the Cumulative variance plot (D) and PCA score plot (E).
Mentions: A total of 1493 raw spectra were obtained between wavelengths (3040-930 cm−1) from all three groups (Fig. 4A). This included 543 from vehicle control, 486 from GNPs-1 and 464 from GNPs-2. This data was further analyzed in the bioband range (1800-930 and 3040-2810 cm−1) (Fig. 4B) followed by the EMSC correction (Fig. 4C) and PCA analysis on the raw data (Fig. 4D,E). PCA was performed on the spectral data of “BioBands” through cross validation at 7 PCs. The interval of the wavelength was kept at 1–60 and 320–548 in the ranges of wave numbers: 3040-2810 cm−1 and 1800-930 cm−1, respectively. The actual bioband wave number was 3039.234 - 2811.677 and 1808.883 - 929.51 cm−1. The specific range was selected since, it is indicated in the identification of lipids, nucleic acid and protein molecules present in the mammalian cell2728.PCA was applied to all the data and provided 3 levels with 95.22% of variance, best observed in three-dimensional space (PC1 versus PC2 versus PC3) (Fig. 4D,E). The PC score plot of unpriced original data showed clear scattering at different PC levels for all the three groups studied.

Bottom Line: Fourier Transform Infrared (FTIR) spectroscopy has become a popular technique in the field of cancer therapy with an ability to elucidate molecular interactions.Towards achieving the aim, we utilized the mouse xenograft model of retinoblastoma and nanoparticle mediated targeted therapy.The results indicate that the mechanism underlying the response differed between the treated and untreated group which can be elucidated by unique spectral signatures generated by each group.

View Article: PubMed Central - PubMed

Affiliation: Department of Nano biotechnology, Vision Research Foundation, Sankara Nethralaya, 18, College Road, Nungambakkam, Chennai - 600 006, India.

ABSTRACT
Rapid monitoring of the response to treatment in cancer patients is essential to predict the outcome of the therapeutic regimen early in the course of the treatment. The conventional methods are laborious, time-consuming, subjective and lack the ability to study different biomolecules and their interactions, simultaneously. Since; mechanisms of cancer and its response to therapy is dependent on molecular interactions and not on single biomolecules, an assay capable of studying molecular interactions as a whole, is preferred. Fourier Transform Infrared (FTIR) spectroscopy has become a popular technique in the field of cancer therapy with an ability to elucidate molecular interactions. The aim of this study, was to explore the utility of the FTIR technique along with multivariate analysis to understand whether the method has the resolution to identify the differences in the mechanism of therapeutic response. Towards achieving the aim, we utilized the mouse xenograft model of retinoblastoma and nanoparticle mediated targeted therapy. The results indicate that the mechanism underlying the response differed between the treated and untreated group which can be elucidated by unique spectral signatures generated by each group. The study establishes the efficiency of non-invasive, label-free and rapid FTIR method in assessing the interactions of nanoparticles with cellular macromolecules towards monitoring the response to cancer therapeutics.

No MeSH data available.


Related in: MedlinePlus