Size Exclusion Chromatography with Multi-Angle Light Scattering (SEC-MALS) for Glycoprotein Analysis

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Efficient And Accurate SEC-MALS Service at Creative Biolabs

SEC-MALS is an analytical technique that combines SEC and MALS. It is widely applied to characterize biomacromolecules, such as proteins, carbohydrates, and nucleic acids. This technique provides information such as biomacromolecules' molecular weight distribution and molecular weight. Creative Biolabs has extensive industry experience in glycoprotein analysis. We have advanced analysis techniques such as HPLC, MS, FTIR, SPRi, microarray, SEC-MALS, etc., which promptly respond to clients' analysis needs and provide clients with high-resolution and accurate analysis results. Creative Biolabs usually performs SEC-MALS analysis on proteins in the following steps.

Sample preparation

Creative Biolabs purifies the obtained glycoprotein samples to remove other impurities that may influence the determination. We then dissolve the samples in an appropriate buffer, commonly used buffers include phosphate-buffered saline (PBS).

SEC-MALS system setup

Column selection: SEC uses the difference in the size of different molecules in the sample for separation. When a sample passes through an SEC column filled with porous fillers, larger molecules cannot enter the pores and are therefore preferentially expelled, while smaller molecules remain in the pores for a longer time. This allows the sample to be separated according to molecular size. Different SEC columns have different pore size ranges. Creative Biolabs selects the appropriate SEC column for analysis according to the size and properties of the target protein. Moreover, the SEC column is equilibrated with appropriate buffers to ensure a stable environment within the column.
Connecting the MALS detector: MALS is a light scattering technique that measures the relationship between the intensity of scattered light and the scattering angle to obtain information about the size and molecular weight of molecules in solution. Creative Biolabs connects the SEC system to the MALS detector and adjusts the angle and sensitivity of the MALS detector to suit the type of sample being analyzed.

Collection and analysis

Creative Biolabs injects the processed protein sample into the SEC-MALS system, monitors the scattered light signal in real time through the MALS detector, and obtains scattering intensity data at different scattering angles. Subsequently, we calculate the molecular weight distribution and molecular weight of the glycoprotein using an appropriate algorithm.

Through the above steps, Creative Biolabs combines the separation capability of SEC with the precise detection of MALS to efficiently and accurately characterize information such as the molecular weight and distribution of glycoproteins. This technology provides important support for biomedical research, especially in the study of the relationship between the structure and functions of glycoproteins. Please contact us to obtain more information if you are very interested in our SEC-MALS analysis services.

High-resolution and accurate SEC-MALS analysis services. (Creative Biolabs Original)

Published data

SEC-MALS is a technique that is particularly well suited for measuring the absolute molar mass of macromolecules, especially proteins in different morphologies. SEC separates proteins by their hydrodynamic radius, while MALS is used to determine the molar mass of the eluted proteins. The combination of these two techniques results in a versatile and reliable method for effectively characterizing the properties of one or more proteins in solution. In this study, the researchers applied SEC-MALS to determine the molar mass of proteins containing [Fe-S] clusters and corrected the light scattering data for errors caused by the absorption of laser light by the sample. The results revealed a relationship between the molar mass changes of the whole NEET protein and its oligomeric state, highlighting the importance of the iron-sulfur cluster in protein function. By combining SEC with MALS, the authors obtained more accurate protein molar mass information without the need for standard calibration, while also being able to analyze the homogeneity of the sample.

Fig.1 SEC-MALS analysis results of proteins.¹

FAQ

Q1: How does the choice of the SEC column affect the analysis results?

A1: The pore size range of the SEC column determines the molecular size that can be separated. Choosing the right SEC column can ensure the effective separation of the target protein in the column, thereby improving the accuracy of the data.

Q2: What specific information can SEC-MALS provide?

A2: SEC-MALS is mainly suitable for the analysis of macromolecules (such as proteins, carbohydrates, and nucleic acids). SEC-MALS can provide information such as molecular weight, molecular weight distribution, polymerization state (such as monomer, dimer, etc.), and relative abundance of different forms.

Q3: What factors affect the accuracy of SEC-MALS analysis results?

A3: The accuracy of SEC-MALS results may be affected by many factors such as sample purity, SEC column selection, flow rate, temperature, and sensitivity of the MALS detector. Therefore, we will strictly control the experimental conditions to ensure reliable results.

Customer Review

Highly Accurate SEC-MALS Analysis Results
"The analysis team of Creative Biolabs is very professional. We cross-validated the analysis results of SEC-MALS and compared them with data from other technologies (such as HPLC and MS). We found that the molecular weight and distribution results provided by SEC-MALS were highly consistent with expectations, laying a solid foundation for our subsequent functional studies."

Advanced Equipment And Fast Analysis Speed
"The efficient performance of the SEC-MALS equipment enabled us to obtain accurate data in a shorter time, which greatly facilitated the time management of our project. The quality and accuracy provided by Creative Biolabs exceeded our expectations and is worth recommending."

Reference

Velours, Christophe, et al. "Determination of the absolute molar mass of [Fe-S]-containing proteins using size exclusion chromatography-multi-angle light scattering (SEC-MALS)." Biomolecules 12.2 (2022): 270. Distributed under Open Access license CC BY 4.0, without modification.

For Research Use Only.