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Related Concept Videos

Raman Spectroscopy: Overview01:20

Raman Spectroscopy: Overview

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The underlying principle of Raman spectroscopy is based on the interaction between light and matter, specifically molecules' inelastic scattering of photons. When a monochromatic beam of light, typically from a laser source, interacts with a sample, most scattered light has the same frequency as the incident light. This is known as Rayleigh scattering.
However, a small fraction of the scattered light exhibits a frequency shift due to the exchange of energy between the incident photons and...
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Chromatographic Methods: Terminology01:18

Chromatographic Methods: Terminology

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Chromatography is an analytical technique widely used in fields such as chemistry, biology, environmental science, and pharmaceuticals to separate the components of a mixture and identify substances between them. The process of chromatography is based on the interactions between two distinct phases: the stationary phase and the mobile phase. The stationary phase is fixed in place by a supporting material, while the mobile phase moves over it, carrying the solutes. As the mobile phase travels,...
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Chromatographic Methods: Classification01:12

Chromatographic Methods: Classification

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Chromatographic techniques are classified in three ways: the classification is based on the physical state of the stationary and mobile phases, how the mobile phase and the stationary phase contact each other, or through the chemical or physical processes that isolate the components of the sample. Typically, the mobile phase is either a liquid or gas, while the stationary phase is either a solid or a liquid layer applied to a solid surface.
Chromatographic techniques are typically named by...
3.9K
Raman Spectroscopy Instrumentation: Overview01:26

Raman Spectroscopy Instrumentation: Overview

1.3K
A conventional Raman spectrophotometer includes a laser source, a sample holding system, a wavelength selector, and a detector.
The monochromatic laser source, typically using visible or near-infrared radiation, generates a highly focused beam of light. This light interacts with the molecules of the sample, scattering some of the light. Liquid and gaseous samples are usually tested in ordinary glass capillaries, while solids can be analyzed as powders packed in capillaries or as potassium...
1.3K
Applications of IR Spectroscopy: Overview01:11

Applications of IR Spectroscopy: Overview

2.3K
The non-destructive nature and ability to provide valuable chemical information make IR spectroscopy a versatile technique with broad applications in various scientific and industrial fields. IR spectroscopy is commonly used to identify and characterize organic and inorganic compounds. It provides information about the functional groups present in a molecule and the bonding between atoms. This helps in the structural elucidation of compounds during organic synthesis, pharmaceutical research,...
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Chromatographic Resolution01:15

Chromatographic Resolution

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In chromatography, a solute moves through a chromatographic column and tends to spread, forming a Gaussian-shaped band. The longer the solute spends in the column, the broader the band becomes. The broadening can lead to overlaps within the column, affecting separation effectiveness.
The effectiveness of separation can be evaluated by determining the level of separation between two neighboring peaks in a chromatogram, which represents the individual components of a sample.
In chromatography,...
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Updated: Feb 3, 2026

Methods to Study Changes in Inherent Protein Aggregation with Age in Caenorhabditis elegans
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Development of Raman Spectroscopy and Machine Learning Methods for Protein Aggregate Quantification: Application to

Jakob Heyer-Müller1, Robin Schiemer1, Lars Robbel2

  • 1Institute of Process Engineering in Life Sciences-Section IV: Biomolecular Separation Engineering, Karlsruhe Institute of Technology (KIT), Fritz-Haber-Weg 2, Karlsruhe, Baden-Württemberg, Germany.

Biotechnology and Bioengineering
|February 2, 2026
PubMed
Summary
This summary is machine-generated.

Raman spectroscopy with machine learning accurately quantifies protein monomers and aggregates in real time. This advanced technique enhances biopharmaceutical manufacturing by enabling rapid detection of protein aggregation, improving product quality and safety.

Keywords:
Raman spectroscopyaggregationchemometricschromatographyconvolutional neural networkprocess analytical technologystructural markers

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Area of Science:

  • Biopharmaceutical Manufacturing
  • Analytical Chemistry
  • Process Analytical Technology

Background:

  • Protein aggregation is a critical issue affecting biopharmaceutical product quality, efficacy, and safety.
  • Traditional offline methods for aggregate detection, like size-exclusion chromatography, lack the speed for real-time process control.
  • There is a significant need for inline analytical techniques to monitor protein monomers and aggregates during manufacturing.

Purpose of the Study:

  • To develop and validate a Raman spectroscopy-based strategy for the selective detection and quantification of protein monomers and aggregates.
  • To address the limitations of traditional methods by enabling real-time monitoring in biopharmaceutical processes.
  • To utilize advanced chemometric approaches for robust quantification of protein size variants.

Main Methods:

  • Raman spectroscopy was employed for molecular specificity and rapid data acquisition.
  • Controlled stress conditions were used to generate reproducible protein aggregates (bovine serum albumin).
  • A Latin Hypercube sampling design varied protein concentration and aggregate fraction to isolate aggregation effects.
  • Convolutional neural networks (CNNs) were utilized as a chemometric machine learning approach for data analysis.

Main Results:

  • Spectral markers indicative of protein aggregation were identified.
  • CNNs demonstrated superior predictive performance and robustness in quantifying monomers and aggregates compared to traditional methods.
  • The developed method achieved reliable, real-time monitoring of protein size variants.
  • Qualitative comparison with offline size-exclusion chromatography validated spectral marker findings.

Conclusions:

  • Raman spectroscopy, coupled with advanced chemometric modeling (specifically CNNs), provides a reliable method for real-time monitoring of protein aggregation.
  • This approach significantly enhances biopharmaceutical downstream process control and robustness.
  • The findings support the application of Raman spectroscopy as a Process Analytical Technology (PAT) for quality control in biopharmaceutical manufacturing.