Monitoring plasma protein aggregation during aging using conformation-specific antibodies and FTIR spectroscopy

Magalhães, Sandra; Trindade, Dário; Martins, Tânia Soares; Rosa, Ilka Martins; Delgadillo, Ivonne; Goodfellow, Brian J.; Silva, Edgar F. da Cruz e; Nunes, Alexandra

doi:10.1016/j.cca.2019.11.025

Cited by 18 publications

(12 citation statements)

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“…FTIR spectroscopy is widely used to assess secondary protein structure in aqueous samples (16)(17)(18) and thus plasma samples were analyzed using this technique to evaluate protein conformational changes across aging. In fact, in our previous work on protein aggregation in human plasma, we verified that the levels of protein aggregates in this biofluid are relatively low (9). Nevertheless, we successfully used FTIR spectroscopy to detect slight changes in protein conformation in samples from donors with different ages.…”

Section: Discussionmentioning

confidence: 64%

“…Fourier Transform Infrared (FTIR) spectroscopy appears as a promising approach for the study of biofluids and has already been successfully used in a clinical setting for diagnosis (8). In our previous work we used FTIR spectroscopy to evaluate age-related changes in protein conformation in human plasma from a dataset of 127 samples (9) and our results suggested that FTIR spectroscopy was able to detect changes in protein secondary structure from middle to old age. When age-associated comorbidities were excluded, a correlation between the age of the donors and secondary structure of plasma proteins was found: samples from younger donors were characterized by antiparallel β-sheetcontaining structures while intermolecular β-sheets characterized older samples (9).…”

Section: Introduction: Proteostasis and Agingmentioning

confidence: 88%

“…In our previous work we used FTIR spectroscopy to evaluate age-related changes in protein conformation in human plasma from a dataset of 127 samples (9) and our results suggested that FTIR spectroscopy was able to detect changes in protein secondary structure from middle to old age. When age-associated comorbidities were excluded, a correlation between the age of the donors and secondary structure of plasma proteins was found: samples from younger donors were characterized by antiparallel β-sheetcontaining structures while intermolecular β-sheets characterized older samples (9). To validate the obtained results and to validate and support the applicability of FTIR as a reliable approach to study proteostasis during aging we have applied the same methodology to a new, independent dataset of human plasma samples to find out if these results corroborate our previous conclusions.…”

Section: Introduction: Proteostasis and Agingmentioning

confidence: 94%

“…For details of FTIR spectral acquisition see section 2.4 of ref (9). Briefly, samples were thawed on ice and homogenized before spectral acquisition in ATR mode in the mid infrared range.…”

Section: Ftir Measurements and Data Analysismentioning

confidence: 99%

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FTIR spectroscopy confirms age-related changes in protein conformation in a new independent dataset of human plasma samples

Magalhães

Teixeira²,

Gouveia³

et al. 2022

MARJ

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The loss of proteostasis is a key hallmark of aging and has been well documented using several model systems. However, in biofluids such as blood plasma, little is known about how the loss of proteostasis affects protein conformation during aging. Our previous work has demonstrated age-related changes in protein conformation in human plasma samples using FTIR spectroscopy and conformation-specific antibodies. To prove the potential of spectroscopy to detect slight changes in protein secondary structure during aging, we have applied the same methodology to a new and independent set of 24 human plasma samples from donors with ages ranging from 40 to 75 years. The results clearly show that from middle to old age there is a decrease in antiparallel β-sheet structures and an increase in intermolecular β-sheets, indicative of an increase in aggregation-prone proteins in human plasma over time. This confirms the potential of FTIR spectroscopy as a technique to evaluate protein conformational changes related to health and disease and as a suitable method to be used in a clinical setting to produce quick and reliable results.

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Section: Discussionmentioning

confidence: 64%

Section: Introduction: Proteostasis and Agingmentioning

confidence: 88%

Section: Introduction: Proteostasis and Agingmentioning

confidence: 94%

“…For details of FTIR spectral acquisition see section 2.4 of ref (9). Briefly, samples were thawed on ice and homogenized before spectral acquisition in ATR mode in the mid infrared range.…”

Section: Ftir Measurements and Data Analysismentioning

confidence: 99%

See 2 more Smart Citations

FTIR spectroscopy confirms age-related changes in protein conformation in a new independent dataset of human plasma samples

Magalhães

Teixeira²,

Gouveia³

et al. 2022

MARJ

Self Cite

View full text Add to dashboard Cite

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“…To calculate the intensity of the spectral bands we used different approaches: for the calculation of intensity of peaks assigned to CH (3013 cm −1 ), CH 2 (2851 cm −1 and 2922 cm −1 ) CH 3 (2959 cm −1 and 2871 cm −1 ), C=O (1741 cm −1 ), glucose (1045 cm −1 ), cholesterol esters (1169 cm −1 ) and protein secondary structures, namely β-sheets (1693 cm −1 , 1682 cm −1 and 1628 cm −1 ), we inverted 2nd derivative correspondent spectra by factoring by −1, as previously described [36,37]. Then we selected the wavenumbers corresponding to that peak and extracted the intensity values.…”

Section: Intensity Of Spectral Bandsmentioning

confidence: 99%

FTIR Spectroscopy as a Tool to Study Age-Related Changes in Cardiac and Skeletal Muscle of Female C57BL/6J Mice

et al. 2021

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Studying aging is important to further understand the molecular mechanisms underlying this physiological process and, ideally, to identify a panel of aging biomarkers. Animals, in particular mice, are often used in aging studies, since they mimic important features of human aging, age quickly, and are easy to manipulate. The present work describes the use of Fourier Transform Infrared (FTIR) spectroscopy to identify an age-related spectroscopic profile of the cardiac and skeletal muscle tissues of C57BL/6J female mice. We acquired ATR-FTIR spectra of cardiac and skeletal muscle at four different ages: 6; 12; 17 and 24 months (10 samples at each age) and analyzed the data using multivariate statistical tools (PCA and PLS) and peak intensity analyses. The results suggest deep changes in protein secondary structure in 24-month-old mice compared to both tissues in 6-month-old mice. Oligomeric structures decreased with age in both tissues, while intermolecular β-sheet structures increased with aging in cardiac muscle but not in skeletal muscle. Despite FTIR spectroscopy being unable to identify the proteins responsible for these conformational changes, this study gives insights into the potential of FTIR to monitor the aging process and identify an age-specific spectroscopic signature.

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Use of spectroscopic process analytical technology for rapid quality evaluation during preparation of CHO cell culture media

Ou,

Cui,

Zhao

et al. 2024

Biotechnology Progress

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Media preparation parameters contribute significantly to media quality, cell culture performance, productivity, and product quality. Establishing proper media preparation procedures is critical for ensuring a robust CHO cell culture process. Process analytical technology (PAT) enables unique ways to quantify assessments and improve media quality. Here, cell culture media were prepared under a wide range of temperatures (40–80°C) and pH (7.6–10.0). Media quality profiles were compared using three real‐time PATs: Fourier‐transform infrared (FTIR) spectroscopy, Raman spectroscopy, and excitation‐emission matrix (EEM) spectroscopy. FTIR and Raman spectroscopies identified shifts in media quality under high preparation temperature (80°C) and at differing preparation pH which negatively impacted monoclonal antibody (mAb) production. In fed‐batch processes for production of three different mAbs, viable cell density (VCD) and cell viability were mostly unaffected under all media preparation temperatures, while titer and cell specific productivity of mAb decreased when cultured in basal and feed media prepared at 80°C. High feed preparation pH alone was tolerated but cell growth and productivity profiles deviated from the control condition. Further, charge variants (main, acidic, basic species) and glycosylation (G0F, afucosylation, and high mannose) were examined. Statistically significant differences were observed for one or more of these quality attributes with any shifts in media preparation. In this study, we demonstrated strong associations between media preparation conditions and cell growth, productivity, and product quality. The rapid evaluation of media by PAT implementation enabled more comprehensive understanding of different parameters on media quality and consequential effects on CHO cell culture.

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Monitoring plasma protein aggregation during aging using conformation-specific antibodies and FTIR spectroscopy

Cited by 18 publications

References 50 publications

FTIR spectroscopy confirms age-related changes in protein conformation in a new independent dataset of human plasma samples

FTIR spectroscopy confirms age-related changes in protein conformation in a new independent dataset of human plasma samples

FTIR Spectroscopy as a Tool to Study Age-Related Changes in Cardiac and Skeletal Muscle of Female C57BL/6J Mice

Use of spectroscopic process analytical technology for rapid quality evaluation during preparation of CHO cell culture media

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