Current Detection Methods of Glycoprotein

Overview of Glycoprotein

Glycoproteins, the family of glycosylated proteins, are a kind of cis-diol-containing biomolecule. Large quantities of glycoproteins are found in nature, including enzymes, antigens, antibodies, and peptide hormones. As we can see, glycoproteins play a vital role in the biological activities of organisms, such as in cell recognition, immunomodulation, and growth regulation. Meanwhile, abnormal glycosylation of proteins in vivo is an important feature in disease progression, being related to the occurrence and development of many diseases, such as infections, tumors, cardiovascular diseases, liver diseases, kidney diseases, diabetes, and certain genetic diseases, so the detection of glycoproteins is of great value in clinical diagnosis, biological research, disease prevention, and so on. The traditional methods of glycoprotein detection, such as mass spectrometry, chromatography-mass spectrometry, and ELISA, are time-consuming, professional, and demanding for sample purity. So in recent studies, researchers developed various novel detection methods.

Current Detection Methods of Glycoprotein

• Fluorescent boronic acid-based molecularly imprinted polymer

In a previous study, a new fluorescent molecularly imprinted polymers (FMIP) were prepared by introducing a fluorescent boronic acid quinoline based on the PGMA/EDMA bead surface and assembled a glycoprotein molecularly imprinted polymer using surface imprinting technology. The FMIP was characterized using Fourier transform infrared (FT-IR) spectroscopy and X-ray photoelectron spectroscopy (XPS). The applications of the fluorescence MIP were further investigated for the determination of template glycoprotein from biological samples under physiological conditions. Researchers used HRP as a template to prepare a fluorescent boronic acid-based MIP by surface imprinting technology. The resultant fluorescent MIP showed high specific selectivity and sensitivity to template glycoprotein when it was used to detect the template HRP from the biological samples under physiological conditions. The approach for the synthesis of the fluorescent MIP provided an alternative solution for the determination of glycoprotein in biological samples, and may also be applied in biological fields.

Fig.1 The synthesis scheme of the fluorescent molecularly imprinted polymers of glycoprotein. (Wei, 2017)

• BAM-based sensors

Boronate affinity materials (BAMs) are a kind of novel adsorbent material based on the properties of specific binding between boronic acid and cis diols, which actively contribute to the recognition, separation, enrichment, or detection of cis-diols. The central principle is that boronic acids can specifically recognize cis-diols and reversibly form five- or six-membered cyclic esters in certain environments. One of the key factors that determined whether boronic acid combines with cis-diols is the pKa of boronic acid. When the surrounding environmental pH is greater than the pKa of boronic acid, cyclic esters are formed, otherwise, cyclic esters are decomposed. Thereby, since the high binding pH is not suitable for the actual physiological detection environment, it is very important to choose an appropriate boronic acid as the ligand. Besides, many parameters need to be taken into comprehensive consideration, including selectivity, affinity, and binding capacity. In addition to the above-mentioned pH-controlled BAMs, temperature-controlled BAMs have also been proposed for the detection of glycoproteins.

Fig.2 Schematic diagram of the interaction between BAMs and glycoproteins. (Qin, 2020)

• Multifunctional oligomer immobilized on quartz crystal microbalance

In another study, researchers developed a novel multifunction oligomer-modified QCM biosensor for glycoprotein. In this work, a multifunctional oligomer was prepared via free radical polymerization and a glycoprotein-imprinted crosslinked film was fabricated directly on the transducer surface of quartz crystal microbalance. Results of template rebinding tests had proven that the cross-linking process film could successfully conserve the orientation of imprinted sites in the aqueous environment and possessed a much higher affinity towards template OVA than that of the non-cross-linked assays. The specific binding affinity was further testified by three distinct control proteins, and the imprinted assay revealed large selectivity factors towards template glycoprotein. This facile and simple strategy holds great promise to fabricate efficient and specific devices for specific tumor marker detection.

Services at Creative Biolabs

With rich experience and advanced technology platform, Creative Biolabs is capable of offering global customers comprehensive glycoprotein-based services including but not limited to:

• Custom Synthesis
• Glycoengineering Service
• Glycoprotein Analysis
• Anti-Glycoprotein Antibody Development
• Glyco-based Vaccine Development

If you are interested in our services or you have any other questions, please don't hesitate to contact us for more information.

References

1. Qin, X.; et al. Boronate affinity material-based sensors for recognition and detection of glycoproteins. Analyst. 2020, 145(23): 7511-7527.
2. Wei, J.R.; et al. Detection of glycoprotein through fluorescent boronic acid-based molecularly imprinted polymer. Anal Chim Acta. 2017, 960: 110-116.

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