Use Of Lectins Research Articles

Lectins as a promising therapeutic agent for breast cancer: A review.

Efficient treatment of cancer has been a subject of research by scientists for many years. Current treatments for cancer, such as radiotherapy, chemotherapy and surgery have been used in traditional combination therapy, but they have major setbacks like non-specificity, non-responsiveness in certain cancer types towards treatment, tumor recurrence, etc. Epidemiological data has shown that breast cancer accounts for 14% of cancer cases occurring in Indian women. In recent years, scientists have started to focus on the use of natural compounds like lectins obtained from various sources to counter the side effects of traditional therapy. Lectins like Sambucus nigra Agglutinin, Maackia amurensis lectin, Okra lectins, Haliclona caerulea lectin, Sclerotium rolfsii lectin, etc., have been discovered to have both diagnostic and therapeutic potential for breast cancer patients. Lectins have been found to have inhibitory effects on various cancer cell activities such as neo-angiogenesis, causing cell cycle arrest at the G1 phase, and inducing apoptosis. The major idea behind the use of lectins in cancer diagnostics and therapeutics is their capability to bind to glycosylated proteins that are expressed on the cell surface. This review focuses on an exploration of the roles of post-translational modification in cancer cells, especially glycosylation, and the potential of lectins in cancer diagnosis and therapeutics.

Development of an Antibody-Based Platform for the Analysis of Immune Cell-Specific N-linked Glycosylation.

N-linked glycosylation plays an important role in both the innate and adaptive immune response through the modulation of cell surface receptors as well as general cell-to-cell interactions. The study of immune cell N-glycosylation is gaining interest but is hindered by the complexity of cell-type-specific N-glycan analysis. Analytical techniques such as chromatography, LC-MS/MS, and the use of lectins are all currently used to analyze cellular glycosylation. Issues with these analytical techniques include poor throughput, which is often limited to a single sample at a time, lack of structural information, the need for a large amount of starting materials, and the requirement for cell purification, thereby reducing their feasibility for N-glycan study. Here, we report the development of a rapid antibody array-based approach for the capture of specific nonadherent immune cells coupled with MALDI-IMS to analyze cellular N-glycosylation. This workflow is adaptable to multiple N-glycan imaging approaches such as the removal or stabilization and derivatization of terminal sialic acid residues providing unique avenues of analysis that have otherwise not been explored in immune cell populations. The reproducibility, sensitivity, and versatility of this assay provide an invaluable tool for researchers and clinical applications, significantly expanding the field of glycoimmunology.

Structural Prediction and Characterization of Canavalia grandiflora (ConGF) Lectin Complexed with MMP1: Unveiling the Antiglioma Potential of Legume Lectins.

A glioblastoma (GBM) is a highly malignant primary brain tumor with a poor prognosis because of its invasiveness and high resistance to current therapies. In GBMs, abnormal glycosylation patterns are associated with malignancy, which allows for the use of lectins as tools for recognition and therapy. More specifically, lectins can interact with glycan structures found on the malignant cell surface. In this context, the present work aimed to investigate the antiglioma potential of ConGF, a lectin purified from Canavalia grandiflora seeds, against C6 cells. The treatment of C6 cells with ConGF impaired the mitochondrial transmembrane potential, reduced cell viability, and induced morphological changes. ConGF also induced massive autophagy, as evaluated by acridine orange (AO) staining and LC3AB-II expression, but without prominent propidium iodide (PI) labeling. The mechanism of action appears to involve the carbohydrate-binding capacity of ConGF, and in silico studies suggested that the lectin can interact with the glycan structures of matrix metalloproteinase 1 (MMP1), a prominent protein found in malignant cells, likely explaining the observed effects.

Supramolecular Binding with Lectins: A New Route for Non-Covalent Functionalization of Polysaccharide Matrices.

The chemical functionalization of polysaccharides to obtain functional materials has been of great interest in the last decades. This traditional synthetic approach has drawbacks, such as changing the crystallinity of the material or altering its morphology or texture. These modifications are crucial when a biogenic matrix is exploited for its hierarchical structure. In this work, the use of lectins and carbohydrate-binding proteins as supramolecular linkers for polysaccharide functionalization is proposed. As proof of concept, a deproteinized squid pen, a hierarchically-organized β-chitin matrix, was functionalized using a dye (FITC) labeled lectin; the lectin used was the wheat germ agglutinin (WGA). It has been observed that the binding of this functionalized protein homogenously introduces a new property (fluorescence) into the β-chitin matrix without altering its crystallographic and hierarchical structure. The supramolecular functionalization of polysaccharides with protein/lectin molecules opens up new routes for the chemical modification of polysaccharides. This novel approach can be of interest in various scientific fields, overcoming the synthetic limits that have hitherto hindered the technological exploitation of polysaccharides-based materials.

Anthelmintic effect of a water soluble Moringa oleifera lectin in rodents experimentally infected with Haemonchus contortus

Allied to the problem of gastrointestinal parasites, especially Haemochus contortus, the use of lectins of plant origin has contributed to the research of alternative anthelmintics. The nematicidal effect of a water soluble Moringa oleifera lectin (WSMoL) was investigated in an experimental model with H. contortus infected Wistar rodents. Three concentrations were tested orally: 5 mg/kg, 2.5 mg/kg and 1 mg/kg. The reduction in the number of larvae recovered in the experimental groups was analyzed, as well as biochemical, hematological and histological parameters. Treatments with 5, 2.5 and 1 mg/kg of WSMoL reduced the number of larvae recovered of animals by 74.7 %, 72.8 % and 66 %, respectively. Untreated infected animals had anemia, moderate mononuclear multifocal hepatitis, vascular congestion in the liver and kidneys, white pulp hyperplasia in the spleen, and presence of eosinophils in the intestine. Infected animals treated with 5 mg/kg of WSMoL showed liver with moderate bleeding, kidney with vascular congestion, spleen with white pulp hyperplasia and intestine with moderate presence of mononuclear cells. An increase in the serum level of glutamic pyruvic transaminase and a reduction in the level of hemoglobin (p < 0.001) were also observed in this group when compared to the uninfected group. However, the administered concentrations of 2.5 and 1 mg/kg of WSMoL were both satisfactory in terms of reducing the number of recovered larvae and not promoting negative changes in the biochemical, hematological and histological parameters evaluated. These results indicate an in vivo nematicidal effect of WSMoL on the H. contortus parasite.

Liposomal phytohemagglutinin: In vivo T-cell activator as a novel pan-cancer immunotherapy.

Immunotherapy is an attractive approach for treating cancer. T‐cell engagers (TCEs) are a type of immunotherapy that are highly efficacious; however, they are challenged by weak T‐cell activation and short persistence. Therefore, alternative solutions to induce greater activation and persistence of T cells during TCE immunotherapy is needed. Methods to activate T cells include the use of lectins, such as phytohemagglutinin (PHA). PHA has not been used to activate T cells in vivo, for immunotherapy, due to its biological instability and toxicity. An approach to overcome the limitations of PHA while also preserving its function is needed. In this study, we report a liposomal PHA which increased PHA stability, reduced toxicity and performed as an immunotherapeutic that is able to activate T cells for the use in future cancer immunotherapies to circumvent current obstacles in immunosuppression and T‐cell exhaustion.

Microscopic Characterization of the Mucous Cells and Their Mucin Secretions in the Alimentary Canal of the Blackmouth Catshark Galeus melastomus (Chondrichthyes: Elasmobranchii)

Sharks belong to the most primitive group of jawed vertebrates and have some special structural and functional features such as a cartilaginous skeleton, a spiral intestinal valve, and a rectal gland for osmoregulation. In January 2020, ten specimens of Galeus melastomus, the Blackmouth catshark, were collected from the Gulf of Asinara (North Sardinia, Italy) and the entire alimentary canal was studied using histochemical reactions to characterize the mucous cell types. In the alimentary canal of G. melastomus, mucous cells mainly secrete a mixture of acidic and neutral mucins. Of the acidic mucins, only the carboxylated type was present in mucous cells of the stomach, while the sulfated type predominated in the esophagus and the intestines. The use of lectins revealed a distribution of sugar residues in mucins related to cellular activities of the different regions of the catshark alimentary canal. The current study is the first report to characterize the intestinal mucous cells of G. melastomus and to provide quantitative data on their different populations in the alimentary canal.

The Use of Lectins as Tools to Combat SARS-CoV-2.

In December 2019, China announced the first case of an infection caused by an, until then, unknown virus: SARS-CoV-2. Since then, researchers have been looking for viable alternatives for the treatment and/or cure of viral infection. Among the possible complementary solutions are lectins, proteins that are reversibly bound to different carbohydrates. The Spike protein, present on the viral surface, can interact with different cell receptors: ACE2, CD147, and DC-SIGNR. Since lectins have an affinity for different carbohydrates, the binding with the glycosylated cell receptors represents a possibility of preventing the virus from binding to the receptors of host cells. In this review, we discuss the main lectins that are possible candidates for use in the treatment of Covid-19, highlighting those that have already demonstrated antiviral activity in vivo and in vitro, including mannose-binding lectin, Griffithsin, BanLec, and others. We also aim to discuss the possible mechanism of action of lectins, which appears to occur through the mediation of viral fusion in host cells, by binding of lectins to glycosylated receptors found in human cells and/or binding of these proteins with the spike glycoprotein, present in virus surface. Moreover, we discuss the use of lectins in clinical practice. Even with the development of effective vaccines, new cases of viral infection with the same virus, or new outbreaks with different viruses can occur; so, the development of new treatments should not be discarded. Moreover, the discussions made in this work are relevant regarding the anti-viral properties of lectins.

Recent Advances in Lectin-Based Affinity Sorbents for Protein Glycosylation Studies.

Glycosylation is one of the most significant post-translational modifications occurring to proteins, since it affects some of their basic properties, such as their half-life or biological activity. The developments in analytical methodologies has greatly contributed to a more comprehensive understanding of the quantitative and qualitative characteristics of the glycosylation state of proteins. Despite those advances, the difficulty of a full characterization of glycosylation still remains, mainly due to the complexity of the glycoprotein and/or glycopeptide mixture especially when they are present in complex biological samples. For this reason, various techniques that allow a prior selective enrichment of exclusively glycosylated proteins or glycopeptides have been developed in the past and are coupled either on- or off- line with separation and detection methods. One of the most commonly implemented enrichment methods includes the use of lectin proteins immobilized on various solid supports. Lectins are a group of different, naturally occurring proteins that share a common characteristic, which concerns their affinity for specific sugar moieties of glycoproteins. This review presents the different formats and conditions for the use of lectins in affinity chromatography and in solid phase extraction, including their use in dispersive mode, along with the recent progress made on either commercial or home-made lectin-based affinity sorbents, which can lead to a fast and automated glycosylation analysis.

Detection of bladder cancer with aberrantly fucosylated ITGA3

We describe a simple, non-invasive assay to identify fucosylated-glycoisoform of integrin alpha-3 (ITGA3) directly from unprocessed urine. ITGA3 was detected directly from the urine of bladder cancer (BlCa) (n = 13) and benign prostatic hyperplasia (BPH) (n = 9) patients with the use of lectins coated on europium-doped-nanoparticles (Eu3+-NPs). Lectin Ulex europaeus agglutinin-I (UEA) showed enhanced binding with BlCa-derived ITGA3. The evaluation with individual samples showed that a glycovariant ITGA3-UEA assay could significantly discriminate BlCa from BPH patients (p = 0.007). The detection of aberrantly fucosylated-isoform of ITGA3 from urine can be used to distinguish BlCa from age-matched benign controls in a simple sandwich assay.

A Simple Biomimetic Receptor Selectively Recognizing the GlcNAc2 Disaccharide in Water.

GlcNAc2 is the core disaccharide fragment present in N‐glycans exposed on the surface of enveloped viruses of high health concern, such as coronaviruses. Because N‐glycans are directly involved in the docking of viruses to host cells, recognition of GlcNAc2 by a biomimetic receptor may be a convenient alternative to the use of lectins to interfere with viral entry and infection. Herein, we describe a simple biomimetic receptor recognizing the methyl‐β‐glycoside of GlcNAc2 in water with an unprecedented affinity of 160 μM, exceeding that of more structurally complex receptors reported in the literature. The tweezers‐shaped acyclic structure exhibits marked selectivity among structurally related disaccharides, and complete discrimination between mono‐ and disaccharides. Molecular modelling calculations supported by NOE data provided a three‐dimensional description of the binding mode, shedding light on the origin of the affinities and selectivities exhibited by the receptor.

A Simple Biomimetic Receptor Selectively Recognizing the GlcNAc2 Disaccharide in Water

AbstractGlcNAc2 is the core disaccharide fragment present in N‐glycans exposed on the surface of enveloped viruses of high health concern, such as coronaviruses. Because N‐glycans are directly involved in the docking of viruses to host cells, recognition of GlcNAc2 by a biomimetic receptor may be a convenient alternative to the use of lectins to interfere with viral entry and infection. Herein, we describe a simple biomimetic receptor recognizing the methyl‐β‐glycoside of GlcNAc2 in water with an unprecedented affinity of 160 μM, exceeding that of more structurally complex receptors reported in the literature. The tweezers‐shaped acyclic structure exhibits marked selectivity among structurally related disaccharides, and complete discrimination between mono‐ and disaccharides. Molecular modelling calculations supported by NOE data provided a three‐dimensional description of the binding mode, shedding light on the origin of the affinities and selectivities exhibited by the receptor.

Changes to the spermatozoa glycocalyx and its role in fertilization in Sauger (Sander canadensis)

The spermatozoa glycocalyx is a dynamic coating of extracellular glycoproteins known to facilitate the acrosome reaction and fertilization in mammals. Most fish sperm, however, contain no acrosome and are subjected to different stimuli (e.g., osmotic shock in the external environment) prior to fertilization. At present, our understanding of the composition and spatial distribution of sugar moieties and the functional role of the glycocalyx in fish sperm remains incomplete. Moreover, the influence of assisted reproduction techniques (e.g., testicular harvest and cryopreservation) commonly used in aquaculture settings on this structure in fish sperm is unknown. Herein, we describe and compare the composition and characteristics of the glycocalyx among sperm types (stripped, testicular, and cryopreserved) and between activation statuses (inactive vs. activated) using sauger (Sander canadensis) as a model. We also investigated the importance of certain moieties (e.g., N-acetyl-glucosamine [GlcNAc]) to fertilization. Staining distributions, fluorescent intensity, and proportion of cells exhibiting high fluorescence were measured for each treatment using fluorescent microscopy and flow cytometry, respectively. Three lectins, specific to certain glycocalyx sugar moieties (wheat germ agglutinin, WGA [GlcNAc]; concanavalin A, ConA [α-mannose]; and peanut agglutinin, PNA [β-galactose]) were used to monitor these variables in fish sperm. The sauger glycocalyx contained GlcNAc and α-mannose but lacked β-galactose moieties. Testicular sperm exhibited fewer cells (20–40% fewer) with high GlcNAc and α-mannose content than other sperm types. Additionally, the proportion of highly fluorescent cells in testicular sperm were positively correlated with motility (r = 0.80–0.95), suggesting the glycocalyx is affected by post-testicular maturation. Motility activation via hypo-osmotic shock caused a redistribution of GlcNAc to the apical region of the head of 40–50% of stripped and testicular sperm, respectively. Cryopreserved sperm showed significantly reduced apical staining following activation (< 5%) and a 2 to 3-fold increase in α-mannose availability. Further, fertilization was reduced by ~80% compared to a fresh control (i.e., untreated stripped sperm) in both stripped sperm pre-treated block GlcNAc during insemination and cryopreserved sperm. These results indicate that the glycocalyx of sauger sperm is dynamic, as it is modified during post-testicular maturation, motility activation, and that it plays a pivotal role in fertilization. Cryopreservation largely negated these changes, which may partially explain the reduced fertility observed in frozen sperm. Ultimately, our analyses show that the use of lectins (e.g., GA and ConA) as biomarkers for the glycocalyx can help to understand the quality and fertilization potential of fish sperm.

Symbiotic Properties of Sinorhizobium meliloti and Ethylene Production by Alfalfa Plants at the Early Stages of the Symbiosis Formation under Different Water Supply and Seed Treatment by Lectin

The symbiotic properties of bacteria significantly influence on the effectiveness of symbiosis and the yield capacity of plants. Therefore, it is important and relevant to study the features of micro- and macrosymbionts interactions, in particular under stressful conditions, and to find ways to improve the productivity of symbiotic systems. Aim. The investigation of the nodulation and nitrogen-fixing activities of Sinorhizobium meliloti as well as the ethylene production by alfalfa plants at the early stages of the formation of symbiotic system under conditions of both optimal and insufficient water supply and the pre-sowing treatment of seeds by lectin. Methods. Microbiological (cultivation of bacteria culture, seed inoculation), physiological (pot experiment), biochemical (nitrogenase activity and ethylene production determination). Results. It was observed a decrease of the nodulation activity of Sinorhizobium meliloti under drought as well as under lectin application, which was accompanied by an increase in the amount of produced by macrosymbiont ethylene at the early stages of the symbiosis formation. At the same time, the nitrogen fixing activity was inhibited at the insufficient water supply only. The use of lectin promoted the symbiotic apparatus functioning under optimal and insufficient water supply. After renewal of irrigation an increase of the nodule weight and nitrogen fixing activity as well as the aboveground mass of alfalfa plants were noted under lectin treatment. Conclusions. It was identified the role of ethylene as a negative regulator of the nodulation processes at pre-sowing treatment of alfalfa seeds with lectin. The formation of the effective nitrogen-fixing system capable of full recovery after drought under lectin application confirms the prospects for further research in the use of lectins to create optimal conditions for the realization of the symbiotic potential of rhizobia and to increase the symbiotic systemˊ resistance to the action of stress factors.

In Ukrainian

The review material belongs to the scientific and practical issues related to the interdisciplinary direction on the border of nanotechnology, surface chemistry and physics, biology and medicine and is based on the use of natural components in the composition of iron-containing bioactive nanocomposites and magnetic fluids in creating effective vector systems for antitumor therapy with minimized side effects on the human body and improved compatibility with other drugs. Such natural components, which have unique properties, significant and not yet realized potential opportunities for practical use, include, in particular, lectins. The aim of the work is to select and analyze the results of works on the extraction of lectins, the study of their properties and application in biology and medicine. Lectins are a group of substances of protein nature (proteins and glycoproteins) of non-immune origin, which have the ability to reversely and selectively bind carbohydrates and carbohydrate determinants of biopolymers without changes in covalent structure and recognize them with extremely high specificity. Due to this property, they are an ideal tool for reading of code in the structure of specific sugar epitopes on the surface of all cells. Lectins are substances of primary synthesis and are present in all kingdoms, types and classes of living organisms. They mediate cellular communication at the molecular level and are involved in many physiological and pathophysiological processes. Pathogenic bacteria and viruses use lectins to attach to the host tissue, which is one of the prerequisites for the development of infection. Blocking of specific pathogen adhesion with lectin inhibitors is the basis of anti-adhesive therapy, an alternative method of treatment of infections caused by multidrug-resistant bacterial strains. Numerous lectins show antitumor activity and are being studied as potential antitumor drugs. To date, they have found practical application in a number of specialized medical fields, such as histology (detection of carbohydrate structures on the surface of cells and tissues), diagnosis of immunodeficiency and chromosomal abnormalities, transplantology (separation of blood cells and lymphoid cells with different antigenic properties). The prospect of use of lectins in the purification of blood from viruses, pathologically altered glycoproteins, in the targeted delivery of drugs to normal or pathologically altered cells and tissues of the body or to infectious agents is considered very significant. The combination of properties of lectins and magnetically sensitive iron-containing nanocomposites in the composition of magnetic fluids for use in oncology is considered relevant and promising.

Anticancer Activity of Lectins from Bauhinia purpurea and Wisteria floribunda on Breast Cancer MCF-7 Cell Lines.

Individual and collaborative efforts are being made worldwide in search of effective chemical or natural drugs with less severe side-effects for treatment of cancer. Due to the specificity and selectivity properties of lectins for saccharides, several plant lectins are known to induce cytotoxicity into tumor cells. To study the antiproliferative activity of two N-acetyl galactosamine specific plant lectins from seeds of Bauhinia purpurea and Wisteria floribunda against MCF-7 Breast cancer cell lines. MTT, lactate dehydrogenase (LDH) leakage, reactive oxygen species (ROS), and caspase- 3 assays and flow cytometry for cell cycle analysis were performed. The agglutinins BPL and WFL; 446 μgml-1 (2.2 μM) and 329 μgml-1 (2.8 μM), respectively caused remarkable concentration-dependent antiproliferative effect on MCF-7. The effect was seen to be a consequence of binding of the lectin to the cell surface and triggering S and G2 phase arrest. Apoptosis induced was found to be associated with LDH leakage, cell cycle arrest and ROS generation. The apoptotic signal was observed to be amplified by activation of caspase-3 resulting in cell death. The study provides a base for detailed investigation and further use of lectins in cancer studies.

Plant-Derived Lectins as Potential Cancer Therapeutics and Diagnostic Tools.

Cancer remains a global health challenge, with high morbidity and mortality, despite the recent advances in diagnosis and treatment. Multiple compounds assessed as novel potential anticancer drugs derive from natural sources, including microorganisms, plants, and animals. Lectins, a group of highly diverse proteins of nonimmune origin with carbohydrate-binding abilities, have been detected in virtually all kingdoms of life. These proteins can interact with free and/or cell surface oligosaccharides and might differentially bind cancer cells, since malignant transformation is tightly associated with altered cell surface glycans. Therefore, lectins could represent a valuable tool for cancer diagnosis and be developed as anticancer therapeutics. Indeed, several plant lectins exert cytotoxic effects mainly by inducing apoptotic and autophagic pathways in malignant cells. This review summarizes the current knowledge regarding the basis for the use of lectins in cancer diagnosis and therapy, providing a few examples of plant-derived carbohydrate-binding proteins with demonstrated antitumor effects.

Lectins as probes of the human intestinal microbiome

The human microbiota is intimately linked to health and disease. However, identifying community members that contribute to a healthy or dysbiotic state remains a challenge. Host factors regulate microbiome composition by promoting colonization or clearance of microbes. One such factor, soluble lectins, can detect microbes at strain‐level resolution by specific recognition of glycans. Several human lectins expressed at mucosal surfaces can bind microbes, but lectin specificity for microbes is under‐characterized. To this end, we have generated and characterized a suite of fluorescently labeled human soluble lectins to elucidate their microbe‐specificity. Using these probes, we analyzed recognition of bacteria from the human intestinal microbiota, and found that different lectins bind distinct populations. These data support the use of lectins as specific probes of microbial glycosylation. Further, analysis of fecal samples using lectins implicated in clearance of pathogens or promotion of colonization could provide a facile method of profiling the microbiota in health and disease.

Use of Lectin-based Affinity Techniques in Breast Cancer Glycoproteomics: A Review.

Changes in glycoprotein content, altered glycosylations, and aberrant glycan structures are increasingly recognized as cancer hallmarks. Because breast cancer is one of the most common causes of cancer deaths in the world, it is highly urgent to find other reliable biomarkers for its initial diagnosis and to learn as much as possible about this disease. In this Review, the applications of lectins to a screening of potential breast cancer biomarkers published during recent years are overviewed. These data provide a deeper insight into the use of modern strategies, technologies, and scientific knowledge in glycoproteomic breast cancer research. Particular attention is concentrated on the use of lectin-based affinity techniques, applied independently or most frequently in combination with mass spectrometry, as an effective tool for the targeting, separation, and reliable identification of glycoprotein molecules. Individual procedures and lectins used in published glycoproteomic studies of breast-cancer-related glycoproteins are discussed. The summarized approaches have the potential for use in diagnostic and predictive applications. Finally, the use of lectins is briefly discussed from the view of their future applications in the analysis of glycoproteins in cancer.

GlycoBIST: A System for Automatic Glycan Profiling of a Target Protein Using Milli-Bead Array in a Tip.

Lectin is a biomolecule that recognizes a specific part of glycans and, thus, has been used widely as a probe for glycoprotein analysis. Owing to the wide repertoire in nature combined with the recent two decades of advances in microarray technology, the multiplexed use of lectins has been widely used for glycan profiling of endogenous proteins. Because protein glycosylation is recognized as being biologically important and is expected to be a reliable disease marker, lectin microarray analysis with highly sensitive detection has been used to discover disease-relevant glycosylation alterations. However, the conventional system is limited to research purposes; thus, its implementation in clinical settings is warranted. Here, we provide an automatic glycan profiling method using GlycoBIST. A unique array format is used for 10-plexed lectin-glycoprotein interaction analysis on 1-mm-sized beads, which are arranged vertically in a capillary-shaped plastic tip. Using a one-boxed autopipetting machine, the whole process (including interaction, washing, and detection) is performed automatically and serially, resulting in reproducible measurements. In this article, a typical method for glycan profiling of a purified glycoprotein and the fabrication of GlycoBIST tips is explained. © 2020 by John Wiley & Sons, Inc. Basic Protocol 1: Fabrication of a GlycoBIST tip Basic Protocol 2: Automatic profiling of a target glycoprotein using GlycoBIST.