Detection Of Sialic Acid Research Articles

Hydrogel Grating Sensors with Boron Affinity and Molecular Imprinting Effects for Rapid and Sensitive Detection of Tumor Marker Sialic Acid.

Rapid and sensitive detection of the concentration of sialic acid (SA) in serum is crucial for early tumor screening and prognostic assessment; however, it still remains challenging. Here, we propose a novel kind of hydrogel grating sensor with boron affinity and molecular imprinting effects (B-MIP) for the rapid and sensitive detection of SA concentration in serum. The hydrogel gratings feature uniform surface relief microstructures and incorporate highly specific recognition binding sites into SA molecules provided by boron affinity and molecular imprinting. The periodic nanoridges of hydrogel gratings increase the specific surface area contacting the environmental solution; therefore, fast detection can be achieved within 2 min. Upon recognition of SA molecules, the height of hydrogel gratings changes at the nanoscale, causing a change in the diffraction efficiency of the hydrogel gratings. The B-MIP hydrogel grating sensors have highly specific binding sites to SA molecules distributed throughout the whole hydrogel and can preferentially and selectively recognize and respond to the SA molecules even in the presence of interference substances glucose and fructose with high concentrations. The B-MIP hydrogel grating sensors are effectively applicable for the rapid and sensitive detection of SA concentrations in real serum samples with satisfactory accuracy and precision. Our approach provides an excellent strategy to address the current challenges in SA detection and provides new insights into the detection of tumor markers in serum, thereby opening up new ways to accurately detect complex biological samples in analytical science.

Sialic acid detection and theranostic activity of phenylboronic acid-based fluorescent probe in human colorectal adenocarcinoma HT-29 cells

A new probe, 4-(((3′,6′-bis(diethylamino)-3-oxospiro[isoindoline-1,9′-xanthen]-2-yl)imino)methyl)phenyl)boronic acid (R4B) was prepared by facile condensation of 4-formylphenylboronic acid and rhodamine B hydrazide. R4B was characterized by spectroscopic methods and single crystal X-ray diffraction. The sensor R4B solution turned pink and emitted orange fluorescence only in the presence of sialic acid but remained colorless and non-fluorescent otherwise. The sugar recognition performance was investigated via UV–vis and fluorescence spectroscopic studies. Our results revealed that R4B has good affinity and selectivity for sialic acid over common monosaccharides, with a detection limit as low as 10−7 M. Furthermore, R4B selectively inhibited growth of human colorectal adenocarcinoma HT-29 (IC50 <20 µM) without significant cytotoxicity to normal human colon fibroblasts CCD-18Co. Treatment with R4B suppressed HT-29 colony formation via mitochondrial apoptosis in a time-dependent manner. Cellular imaging studies also revealed the ability of R4B as a fluorescence dye to detect intracellular sialic acid and showed mitochondria-tracking ability in HT-29 cells. In summary, R4B is a potential theranostic for the detection of intracellular sialic acid during the early incubation period, followed by induction of cancer apoptotic cell death at a later treatment point.

Surface Sialic Acid Detection of Small Extracellular Vesicles at the Single-Particle Level by Nano-Flow Cytometry.

Glycans, particularly sialic acids (SAs), play crucial roles in diverse biological processes. Despite their significance, analyzing specific glycans, such as sialic acids, on individual small extracellular vesicles (sEVs) has remained challenging due to the limited glycan capacity and substantial heterogeneity of sEVs. To tackle this issue, we introduce a chemical modification method of surface SAs on sEVs named PALEV-nFCM, which involves periodate oxidation and aniline-catalyzed oxime ligation (PAL), in conjunction with single-particle analysis using a laboratory-built nano-flow cytometer (nFCM). The specificity of the PALEV labeling method was validated using SA-decorated liposomes, enzymatic removal of terminal SA residues, lectin preblocking, and cellular treatment with an endogenous sialyltransferase inhibitor. Comprehensive mapping of SA distributions was conducted for sEVs derived from different sources, including conditioned cell culture medium (CCCM) of various cell lines, human saliva, and human red blood cells (RBCs). Notably, treatment with the calcium ionophore substantially increases the population of SA-positive RBC sEVs and enhances the SA content on individual RBC sEVs as well. nFCM provides a sensitive and versatile platform for mapping SAs of individual sEVs, which could significantly contribute to resolving the heterogeneity of sEVs and advancing the understanding of their glycosignature.

Self‐assembly imprinting fabrication of shell‐crosslinking fluorescent nanoparticles for the detection of sialic acid

AbstractA new strategy for preparing sialic acid (SA)‐quenching fluorescent nanoparticles by macromolecular self‐assembly and molecular imprinting technique is suggested. An amphiphilic block copolymer polystyrene‐b‐poly(glycidyl methacrylate) (PS‐b‐PGMA) was designed and synthesized. After functionalization, PS‐b‐PGMA was co‐assembled with the template molecule SA to prepare SA‐imprinted fluorescent nanoparticles, named as nano‐FMIPs. In this process, the shell layers of nanoparticles were photo‐crosslinked upon 365 nm UV light to stabilize the recognition site. The size of the crosslinked shell was found to be increased to 420 ± 5 nm by TEM and DLS measurements. After extraction of SA, recognition cavities complementary in shape, size, and chemical function toward the template SA were left on the surface of nano‐FMIPs, which can selectively rebind the target SA in a mixture of structurally similar compounds. It is found that SA can significantly quench the fluorescence intensity of nano‐FMIPs linearly within a concentration range of 1 × 10−2 ~ 2 × 10−6 mol L−1 and a detection limit of 1.55 × 10−6 mol L−1. This strategy provides a new approach for preparing surface‐imprinted nano‐FMIPs for the quantitative detection of SA, and the nano‐FMIPs may have great potential for biomedical application.

Development of Quantifiable Assays for Detection of Hypersialylation in Oral Cancer Diagnosis

Aim: The aim of this study is to Quantify sialic acid for oral cancer diagnosis using different method using densitometric analysis. Background: Early detection of OSCC is critical for improving survival rates and reducing morbidity, which has prompted researchers to look for potential biomarkers to aid in early diagnosis. Saliva is an excellent diagnostic tool due to its noninvasive nature. Sialic acid levels in saliva have been found to be higher in cancer patients. Hypersialylation is associated with poor prognosis and accelerated cancer progression, as well as immune escape, tumor growth and spread, angiogenesis, and resistance to apoptosis and treatment. This study addresses the imperative need for adaptive, effective, and non-invasive strategies in the diagnosis and treatment of oral cancer, emphasising the importance of early detection for improved prognosis. This study introduces various methods for quantify levels of sialic acid in saliva, including resorcinol and orcinol assays, using colorimetric and densitometric assays. Material and Method: In this study, sialic acid detection methods have been standardized using standards as well as spent media from cancer cell lines to develop noninvasive diagnostic tools for early detection. Two methods used for the detection of sialic acid Resorcinal and Orcinol method by densitometric analysis using Image J software. Results: The study has successfully quantified sialic acid from spent media and saliva using resorcinol and orcinol method by densitometric analysis. Conclusion: In this study we aim to develop a reliable and cost-effective method for quantification of sialic acid using Image j software. The approach presents a non-invasive and efficient alternative for early cancer diagnosis showcasing its potential impact on advancing diagnostic technologies.

Self-enhanced luminescence of copper nanowires combined with pH-sensitive glycosyl imprinted polymers: Electrochemiluminescence for high-sensitivity detection of sialic acid

It is of great significance to explore a low cost and high sensitivity sialic acid (SA) detection method for early diagnosis and prognosis of cancer. Electrochemiluminescence (ECL) has aroused many scholars' interest in the accurate analysis of multi-domain analytes for its excellent performance. In order to improve the ECL signal strength and the corresponding detection sensitivity, it is effective to develop new ECL reagents. Copper nanowires (CuNWs) as luminophor has always been a blank in the field of ECL sensors. In this study, a glycosyl imprinted electrochemiluminescence sensor (GIP-ECLS) for SA specificity detection in serum was developed using CuNWs as luminophor. CuNWs can not only form quasi-three-dimensional (3D) conductive network, but also realize self-enhancement of ECL signal through Cu2+ and Cu+ conversion. In addition, the excellent conductivity and wrapping properties of Ti3C2 provide a large electroactive area for electrode modification. More importantly, 3-aminobenzenoboric acid (3-ABA) specifically binds to SA to form B-N or B-O, which lays the foundation for selective recognition of SA. Under optimal conditions, the signal response of GIP-ECLS showed a good linear relationship with SA concentration at 5 × 10−12–1 × 10−8 mol L−1 (R2 = 0.997), and the limit of detection (LOD) was 1.17× 10−12 mol L−1. The results showed that in the actual sample analysis, the recovery rate of SA detected by GIP/Ti3C2/CuNWs/GCE was good, and it had good selectivity and stability, which provided ideas for designing minimally invasive and highly sensitive detection of SA in serum samples.

Synthesis of Bone Meal-derived 4-Carboxyphenylboronic Acid Functionalized Sulfur and Nitrogen Co-doped Graphene Quantum Dots Nanoprobe for Sialic Acid Sensing

Detection of sialic acid using advanced sensors in milk-based products is essential in the food industry. Therefore, the present work reports the sulfur and nitrogen-doped graphene quantum dots from bone meal functionalized with boronic acid (Boro-S/N-dGQDs) nanoprobe for sialic acid sensing applications. Briefly, S/N-dGQDs were functionalized with 4-carboxyphenylboronic acid to improve performance of fluorescent sensors toward the detection of sialic acid. Here, boronic acid surface decoration on S/N-dGQD was confirmed by several spectral characterizations. The addition of different quantities of sialic acid results in a directly proportionate correlation to fluorescence quenching. It gives a broad linear range of 50 ng/mL to 1000 ng/mL and a limit of detection of 6.04 ng/mL. Also, it displayed remarkable selectivity, likely due to interaction of sialic acid-containing 1,2-diol with hydroxyl group of Boro-S/N-dGQDs nanoprobe. Designed sensor demonstrated good stability and reproducibility. Real-time analysis of sialic acid in different milk-based products confirmed practicability of Boro-S/N-dGQDs.

Simple Binding and Dissociation of a Sialoglycoprotein Using Boronic Acid-Modified Functional Interfaces on Microparticles.

An overexpression of sialic acid is an indicator of metastatic cancer, and selective detection of sialic acid shows potential for cancer diagnosis. Boronic acid is a promising candidate for this purpose because of its ability to specifically bind to sialic acid under acidic conditions. Notably, the binding strength can be easily modulated by adjusting the pH, which allows for a simple dissociation of the bound sialic acid. In this study, we developed 5-boronopicolinic acid (5-BPA)-modified magnetic particles (BMPs) to selectively capture sialic acid biomolecules. We successfully captured fetuin, a well-known sialoglycoprotein, on BMPs at >104 molecules/particle using an acetate buffer (pH 5.0). Facile dissociation then occurred when the system was changed to a pH 7.6 phosphate buffer. This capture-and-release process could be repeated at least five times. Moreover, this system could enrich fetuin by more than 20 times. In summary, BMPs are functional particles for facile purification and concentration through the selective capture of sialic acid proteins and can improve detection sensitivity compared with conventional methods. This technology shows potential for the detection of sialic acid overexpression by biological particles.

Electrochemiluminescence Analysis of Multiple Glycans on Single Living Cell with a Closed Bipolar Electrode Array Chip.

The profiling of multiple glycans on a single cell is important for elucidating glycosylation mechanisms and accurately identifying disease states. Herein, we developed a closed bipolar electrode (BPE) array chip for live single-cell trapping and in situ galactose and sialic acid detection with the electrochemiluminescence (ECL) method. Methylene blue-DNA (MB-DNA) as well as biotin-DNA (Bio-DNA) codecorated AuNPs were prepared as nanoprobes, which were selectively labeled on the cell surface through chemoselective labeling techniques. The individual cell was captured and labeled in the microtrap of the cathodic chamber, under an appropriate potential, MB molecules on the cellular membrane underwent oxidation, triggering the reduction of [Ru(bpy)3]2+/TPA and consequently generating ECL signals in the anodic chamber. The abundance of MB groups on the single cell enabled selective monitoring of both sialic acid and galactosyl groups with high sensitivity using ECL. The sialic acid and galactosyl content per HepG2 cell were detected to be 0.66 and 0.82 fmol, respectively. Through comprehensive evaluation of these two types of glycans on a single cell, tumor cells, and normal cells could be effectively discriminated and the accuracy of single-cell heterogeneous analysis was improved. Additionally, dynamic monitoring of variations in galactosyl groups on the surface of the single cell was also achieved. This work introduced a straightforward and convenient approach for heterogeneity analysis among single cells.

Detection of Sialic Acid to Differentiate Cervical Cancer Cell Lines Using a Sambucus nigra Lectin Biosensor.

Pap smear screening is a widespread technique used to detect premalignant lesions of cervical cancer (CC); however, it lacks sensitivity, leading to identifying biomarkers that improve early diagnosis sensitivity. A characteristic of cancer is the aberrant sialylation that involves the abnormal expression of α2,6 sialic acid, a specific carbohydrate linked to glycoproteins and glycolipids on the cell surface, which has been reported in premalignant CC lesions. This work aimed to develop a method to differentiate CC cell lines and primary fibroblasts using a novel lectin-based biosensor to detect α2,6 sialic acid based on attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) and chemometric. The biosensor was developed by conjugating gold nanoparticles (AuNPs) with 5 µg of Sambucus nigra (SNA) lectin as the biorecognition element. Sialic acid detection was associated with the signal amplification in the 1500-1350 cm-1 region observed by the surface-enhanced infrared absorption spectroscopy (SEIRA) effect from ATR-FTIR results. This region was further analyzed for the clustering of samples by applying principal component analysis (PCA) and confidence ellipses at a 95% interval. This work demonstrates the feasibility of employing SNA biosensors to discriminate between tumoral and non-tumoral cells, that have the potential for the early detection of premalignant lesions of CC.

Molecule-induced n-type behavior of phosphorene-based field-effect transistor for highly sensitive detection of sialic acid

Low-concentration detection of sialic acid (SA) seems to be a promising method in early cancer prognosis. In this work, a phosphorene-based field-effect-transistor (Ph-FET) device has been introduced for detecting various concentrations of SA. The device was fabricated using successive hydrogen plasma treatment to start from the red phosphorus layer and to realize highly crystalline black phosphorus sheets. The morphology and crystalline quality of phosphorene sheets have been extensively examined by transmission electron microscopy, scanning electron microscopy, atomic force microscopy, and Raman spectroscopy. By utilizing this device, we can detect different concentrations of SA from 3 to 2400 μM owing to its negative charge. The detection limit of SA is as low as 2.8 μM. This technique offers a straightforward method for the fabrication of high-performance Ph-FET with no need for exfoliation or sheet transfer. We have fabricated a Ph-FET with a mobility of 10 cm2 V−1 s−1, a carrier concentration of 3.9 × 1011 cm−2, and an on/off current ratio of 103. Moreover, we have observed that exposure to SA solution would change the carrier type of the transistor from a p-type channel into an n-type channel device due to the interaction of SA molecules with the phosphorene atoms at the top of the puckered structure that weakens the P-P lattice anti-bonding orbitals and screens the trapped sites.

Detection of Sialic Acid and Imaging of Cell-Surface Glycan Using a Fluorescence-SERS Dual Probe.

Sialic acid (SA) is an acidic monosaccharide present in the human brain and body fluids in the form of N-acetylneuraminic acid. It is also a well-known cancer biomarker. For decades, it has remained a challenging task to design synthetic receptors for SA. However, mainly because of the interference from other sugars with the receptors, it was challenging to differentiate SA from other sugars. Here, we report the development of a two-component aggregation-induced emissive (AIE) probes that can interact with SA and other saccharides via noncovalent interactions with unique emission fingerprints. Analysis of the output signals enabled the reliable detection and clear discrimination of SA in the presence of other saccharides with high accuracy. Further, its potential application in cellular glycan mapping has been explored by fluorescence imaging and surface-enhanced Raman scattering with MDA-MB-231 breast cancer cells.

Highly sensitive and real-time detection of sialic acid using a solution-gated graphene transistor functionalized with carbon quantum dots

Sialic acid (SA) involves multiple different physiological and pathological processes. The SA level in serum is a reliable biomarker for the early diagnosis of malignant tumors. In this study, a new type of solution-gated graphene transistor was designed and fabricated for highly sensitive and real-time detection of SA by rapid binding of boric acid groups to diol monosaccharides. The gate electrode was modified with synthesized carbon quantum dots having boronic acid on the surface as the specific recognition site of SA. The detection of SA is achieved via the change of the gate surface potential after the conjugation of SA with carbon quantum dots. The device sensitivity can reach 10−19 M and a good linear relationship was present between the electrical current response and logarithmic values of the SA concentrations in the range of 10−19 M to 10−5 M. This sensor also exhibited good selectivity and good stability. Furthermore, the detection of SA in simulated serum samples was also carried out and satisfactory results were acquired, showing the high potential of the prepared sensor in clinical application.

Tumor identification via in vivo portable Raman detection of sialic acid with a dual gold nanoprobe system.

A dual gold nanoprobe system was designed for in vivo portable Raman detection of sialic acid (SA) for tumor identification. The dual gold nanoprobe system contained two gold nanoprobes, Au10-DTTC/PEG-PBA and Au40-PEG-SA. Au10-DTTC/PEG-PBA was constructed on a 10 nm gold nanoparticle modified with 3,3'-diethylthia tricarbocyanine iodide (DTTCI) as the Raman reporter and 3-aminophenylboronic acid (APBA) through a thiol PEG succinimidyl carboxymethyl ester (HS-PEG-NHS) linker for specific recognition of SA. Au40-PEG-SA was constructed on a 40 nm gold nanoparticle modified with SA through HS-PEG-NHS. For in vivo detection of SA, Au10-DTTC/PEG-PBA and Au40-PEG-SA were subsequently injected into tumor xenografted mice with optimal interval and retention times. Through the specific recognition between PBA and SA, the conjugates of Au10-DTTC/PEG-PBA and Au40-PEG-SA formed in the tumor region emitted strong SERS signals of DTTC, which could be detected by a portable Raman detector. This work provides a convenient and portable method to detect SA in tumor xenografted mice, which is useful for family-stay identification and clinical cleavage of tumors.

Oxidase mimicking Co/2Fe MOF included biosensor for sialic acid detection

A facile amperometric biosensor that included oxidase mimicking Co/2Fe metal-organic framework (MOF) for sialic acid (SA) detection was prepared. Amperometric SA biosensor was constructed on a gold screen-printed electrode via immobilization of Co/2Fe MOF and N-acetylneuraminic Acid Aldolase (NANA-Aldolase) enzyme, respectively. NANA-Aldolase enzyme converts free SA into pyruvate and N-acetyl-d-mannosamine. After this conversion, oxidase mimicking Co/2Fe bimetallic MOF converts pyruvate into acetylphosphate and O2 into H2O2. Investigation of analytical characteristics resulted with the linear range of 0.02 mM–1.00 mM of SA concentration with limit of detection value of 0.026 mM. Sample application studies with developed SA biosensor were carried out with GD3 ganglioside and HeLa cancer cell lines which have high SA concentrations while A549 cell lines were also used as control group. Before detecting free SA, the bound SA was freed from SA sources where every step was monitored via electron impedance spectroscopy. Then, free SA was successfully detected with the amperometric SA biosensor and as a result, more practical and accurate system was developed.

Layer-by-layer assembly of composite conductive fiber-based organic electrochemical transistor for highly sensitive detection of sialic acid

A novel composites conductive fiber based on Poly(diallyldimethylammonium chloride) and multi-walled carbon nanotubes/Poly(3, 4-ethylenedioxythiophene)-Poly(styrenesulfonate) has been prepared using layer-by-layer assembly (LBL) technology. The characterization results showed the composites film was successful prepared on the cotton fiber and had flexibility, conductivity and washability. The fiber-based organic electrochemical transistors (FECTs) have been assembled by the LBL composite fiber, which exhibit an on/off ratio of 5.12*102±17.99, a peak transconductance value of 8.7 mS, a switching response time of 1.2 s, and excellent cycle stability. Bending and repetitive hand-washing cycles of fibers have no significant influence on the electrical properties of FECTs. Moreover, FECTs maintain stable electrical properties within a week. The gate electrode of the FECT was modified with 3-aminophenylboronic acid to prepare a sialic acid (SA) biosensor. The SA biosensor based on FECTs presented a sensitivity of 57.66 mV per decade, a good linear response in a wide concentration range of 1 nM-1 mM, a linear coefficient R2=0.9848. This biosensor can be successful used to detect the SA content in saliva. Most importantly, the sensor can be integrated into fabrics with no significantly change in the device performance before and after weaving, providing an efficient detection platform for wearable healthcare application.

Colorimetric-assisted photoelectrochemical sensing for dual-model detection of sialic acid via oxidation-power mediator integration

Development of sensitive and accurate methods for sialic acid (SA) determination is of great significance in early cancer diagnosis. Here, a colorimetric-assisted Photoelectrochemical (PEC) sensor was constructed for SA detection based on the two pairs of cis-diol groups in SA molecule. With the specific recognition of SA via the two pairs of cis-diol groups, the prepared gold-modified Bi2S3 (Au [email protected]2S3) and metal organic framework ([email protected]) were introduced to the electrode and formed a sandwich structure. Based on the properties of inert electroconductivity and nanozyme of the PCN-224, dual-readout of photocurrent and visualization was achieved with the presence of 3,3′,5,5′-tetramethylbenzidine (TMB). Moreover, to intensify the visualization signal, Ce3+ was employed as the mediator to boost the catalysis capability by transferring energy from PCN-224 surface to the whole system. With the unique SA recognition and the mediation of Ce3+, both the photocurrent and the visualization signals sensitively responded with the SA concentration linearly. The present method showed greatly high sensitivity, selectivity and accuracy for SA detection with a limit of detection of 1.44 μmol/L and a wide linear range of 5-1000 μmol/L. This method provided a new promising platform for SA detection and a potential strategy for design of novel biosensors with dual-model readout.

Clinical application of liver diseases diagnosis using ultrahigh-sensitive liquid chromatography-mass spectrometry for sialic acids detection

Sialic acids are a group of nine-carbon N-acetylated derivatives of neuraminic acid containing a keto group at position C2 (ketononose), which play important roles in many biological processes. The simultaneous detection of free sialic acid (FSA) and total sialic acid (TSA) is always challenging due to three orders of magnitude difference. An accurate and robust chemical derivatization-based liquid chromatography-mass spectrometry (LC-MS) method was proposed here to quantify both TSA and FSA in human serum with only 1 μL human serum consumption. The derivatization method with Girard's P (GP) reagent provided an ultrasensitive analysis of sialic acids as only [GP+SA-H2O]+ ions derivatized from SA could be detected by LC-MS. The limit of quantification (LOQ) of SA was less than 5 fg (S/N = 47), which was the most sensitive measurement published for SAs in biomatrices. In addition, no matrix effect existed after 10000-fold dilution of serum extracts. The recovery rates were in the range of 98.1–114.0% and the coefficient of variations (CV) was within 5%. The method has been successfully applied for the quantification of TSA and FSA in serums of patients with different liver diseases. The specificities of TSA and FSA for the early diagnosis of severe hepatopathies were higher than most of the lab blood test indicators with area under the curve (AUC) of 0.900 and 0.882.

Detection of Sialic Acids on the Cell Surface Using Flow Cytometry.

We described a method to detect α2-3 linked and α2-6 linked sialic acids on the cell surface with using flow cytometry. Cells were fixed with 4% paraformaldehyde, and then α2-3 and α2-6 sialic acids were stained with biotinylated MAACKIA AMURENSIS LECTIN II (MALII) and biotinylated ELDERBERRY BARK LECTIN (SNA), respectively. Sialic acids on the cell surface were cleaved by sialidase in acetate buffer at pH5.5 to confirm the specificity of staining. Streptavidin conjugated with Alexa flour 488 was used to detect biotinylated lectins. Thus, the α2-3 linked and α2-6 linked sialic acids on the cell surface were semi-quantitatively detected by flow cytometry.

Cathodic Detection of Sialic Acid by a Self-Powered Molecularly Imprinted Photoelectrochemical Sensing Platform Employing Wo3/Bi2s3photoanode and Cuins2 Photocathode

Cathodic Detection of Sialic Acid by a Self-Powered Molecularly Imprinted Photoelectrochemical Sensing Platform Employing Wo3/Bi2s3photoanode and Cuins2 Photocathode