News from LabRulezICPMS Library - Week 39, 2025

Our Library never stops expanding. What are the most recent contributions to LabRulezICPMS Library in the week of 22nd September 2025? Check out new documents from the field of spectroscopy/spectrometry and related techniques!

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This week we bring you application notes by Agilent Technologies and Shimadzu and poster by Thermo Fisher Scientific!

1. Agilent Technologies: Protein Glycosylation Analysis Using the Agilent Cary 630 FTIR Spectrometer

• Application note
• Full PDF for download

Protein glycosylation is a crucial post-translational modification (PTM) impacting the efficacy, stability, and integrity of therapeutic proteins, including monoclonal antibodies (mAbs) and other biopharmaceuticals. This process involves attaching sugar molecules, or glycans, to specific amino acid residues on proteins. Glycosylation is classified as a critical quality attribute (CQA) and has important functions in biological processes, impacting protein structure, solubility, and ability to resist proteolytic degradation.1 Therefore, monitoring and controlling glycosylation is essential in developing and producing therapeutic proteins.

Various analytical techniques are employed to monitor glycosylation in biopharmaceutical production. These techniques include FTIR spectroscopy, mass spectrometry, high-performance liquid chromatography, capillary electrophoresis, nuclear magnetic resonance spectroscopy, and lectin arrays. However, most of these techniques require time-consuming complex sample preparation steps. Alternatively, FTIR is a robust analytical technique that provides a fast and easy tool for protein research. It is a nondestructive technique used to study protein molecules under different experimental conditions. The building blocks of glycans (monosaccharide/sugars), amide groups of protein, and water molecules produce distinguishable fingerprint absorption bands in FTIR. It has also been used to monitor the carbohydrate content and glycosylation profiles in glycoproteins.2,3 

In this study, we used an Agilent Cary 630 FTIR spectrometer to demonstrate an FTIR spectroscopy method for protein glycosylation analysis. The results show that FTIR spectra of glycoproteins provide an FTIR signal corresponding to the glycosylation profile, allowing a rapid, nondestructive analysis method with minimal sample preparation.

Experimental 

Instrumentation 

The Cary 630 FTIR spectrometer was fitted with a single reflection diamond attenuated total reflection (ATR) module. Data acquisition was carried out with an Agilent MicroLab software using the parameters shown in Table 1.

Results and discussion 

Proteins exhibit characteristic amide absorption bands in FTIR spectroscopy, which are related to secondary structure and confirmation. The amide I band (1,600 to 1,800 cm–1) and amide II band (1,470 to 1,570 cm–1) result from the absorption of the carbonyl and N-H group. 

Glycosylation is one of the common PTMs that occur in biopharmaceuticals. Carbohydrates linked to glycoproteins have an absorption band distinct from the protein amide region. The infrared (IR) absorption of glycan spans broadly in the 1,200 to 950 cm–1 region where proteins have very low signal intensity. It has been demonstrated that the amount of carbohydrates in the glycoprotein correlates with the intensity of these spectral bands.3 

In this study, various glycosylated proteins with different levels of glycosylation were selected to evaluate the FTIR method. The formulated mAbs contain excipients that can exhibit absorption bands, and these signals may interfere with glycan analysis in the 1,200 to 950 cm–1 region. Therefore, mAb samples were buffer-exchanged with water using spin columns. Figure 3 shows the comparison of FTIR spectra of formulated and buffer-exchanged mAbs. The desalting step is necessary to eliminate buffer interference with the glycosylation measurement.

Conclusion 

This application note demonstrates the simplicity and usability of glycosylation analysis using an Agilent Cary 630 FTIR spectrometer. The FTIR method provides a spectral variation related to the global glycosylation level of protein, removing the necessity for extensive sample preparation or labeling. It provides a signal corresponding to the glycan profile that allows the identification of glycoproteins and enables rapid comparative glycosylation analysis between innovator and biosimilar therapeutic proteins. The user‑friendly Agilent MicroLab Expert software streamlines data processing and shortens time to results. The technique may be used to verify batch consistency and product quality during the protein production process. Its capability for quick glycosylation estimation makes it an excellent fit for protein quality control workflows. In summary, the FTIR approach provides numerous advantages, including minimal sample preparation, reduced analysis time, and the capacity to analyze intact glycoproteins

2. Shimadzu: Peel Test for Cathode Materials of Lithium-Ion Batteries in a Thermostatic Chamber

• Application note
• Full PDF for download

User Benefits

• A thermostatic chamber makes it possible to measure the adhesion force of cathode materials in actual temperature environments. 
• A copper foil peeling test device enables to conduct peel tests while maintaining a 90° angle in a thermostatic chamber. 
• By using a load cell with a wider accuracy guarantee range than a conventional one, it is possible to conduct high-accuracy tests even with small test forces.

Lithium-ion batteries play an important role in various electronic devices due to their high energy density and excellent charging efficiency. To improve battery performance, materials and processing methods are actively being developed, and strength measurement is one of the essential evaluation methods. This Application News focuses on the adhesion strength of the electrode sheets used as battery electrodes. Fig. 1 shows the structure of an electrode sheet. An electrode sheet consists of an electrode material that adheres to a current collector, through which electrons move. The electrode material is composed of a cathode active material such as lithium iron phosphate and an insulating material known as a binder. When the adhesion between the electrode material and the current collector is weak, the electrode material may delaminate from the current collector, preventing electrons from moving in the electrode sheet and increasing the electrical resistance. Therefore, evaluating the adhesion strength is important for improving battery performance. In this application, test equipment, test conditions, and results of the peel test for battery cathode materials are described. The temperature dependence of the adhesion strength of the electrode materialsis also described.

• Testing Machine: Precision Universal Testing Machine AGS-V

Conclusion 

Peel tests in a thermostatic chamber were conducted using an AGS-V and TCE-N300A for Lithium iron phosphate used as cathode sheet of lithium-ion batteries. From the test results, it was confirmed that the adhesion force between the cathode active material and the aluminum foil increased as the temperature increased. This measurement can easily evaluate the temperature dependence of mechanical characteristics of battery cathodes. This method is expected to be applied to solve problems for improving battery performance.

3. Thermo Fisher Scientific: Assessing the level and distribution of trace elements in individual cells using single cell ICP-MS analysis

• Poster
• Full PDF for download

Trace elements play a key role in many processes involved in living beings. Some elements are found as constituent elements in biopolymers, such as phosphorous as part of the DNA backbone, or sulfur, as part of the key amino acids methionine and cysteine involved in the formation of proteins covering a wide variety of functions. Other elements (in particular, the transition elements copper, iron, and zinc) are involved in specific functions, such as acting as co-factors in enzymes. However, the distribution of trace elements cannot be assumed to be homogeneous across a cell cohort, even under ideal conditions. Ultimately, cell-tocell variability of the metal content may be an important factor in understanding biological diversity. The analysis of specific biomarkers at the single cell level has therefore gained significant interest in recent years. 

The analysis of the content of a given metal at the single cell level has remained a challenge. While inductively coupled plasma mass spectrometry (ICP-MS) is well known as a powerful and element-selective detection system, it is only recently that its capacity for analyzing individual cells has been recognized.

Methods

A triple quadrupole ICP-MS system, in conjunction with the scQuant plug-in for Thermo Scientific™ Qtegra™ Intelligent Scientific Data Solution software was used for all measurements.

Conclusions 

This poster highlights how ICP-MS operated in the single cell mode can determine the amount of different trace elements in individual cells and can therefore allow not only assessment of the average element mass per cell, but also the element distribution across the cohort. 

• Scanning for multiple elements is possible using a sequential approach, in which all the elements of interest are measured for an identical period in a single aspiration of a sample, and the results are summarized in a single data set. 
• The scQuant plug-in allows integrated control over sample delivery devices, in this case, a syringe pump, to facilitate a stable sample flow at the required low flow rates. 
• Key method parameters, such as detection sensitivity and transport efficiency can be determined as part of the same Qtegra ISDS Software LabBook or can be independently determined (if no suitable standards are available) and applied to a data set. 
• The data visualization features of the scQuant plug-in allow raw data and intermediate data (signal distribution) to be comprehensively evaluated, as well as enabling representation of results in either a histogram or a box plot. All results can be exported in a spreadsheet compatible format if required.