Development of a Lectin Microarray for the Rapid Analysis of Protein Glycopatterns

Pilobello, Kanoelani T.; Krishnamoorthy, Lakshmipriya; Slawek, Deepika; Mahal, Lara K.

doi:10.1002/cbic.200400403

Cited by 289 publications

(232 citation statements)

References 33 publications

Supporting

Mentioning

227

Contrasting

Order By: Relevance

“…Recently, several reports on lectin arrays have appeared in the literature [41][42][43][44]. In all of the approaches, lectins are arrayed onto glass slides, and the binding of either intact glycoproteins [41][42][43] or whole cells [44] is measured.…”

Section: Discussionmentioning

confidence: 99%

“…In all of the approaches, lectins are arrayed onto glass slides, and the binding of either intact glycoproteins [41][42][43] or whole cells [44] is measured. The major advantages of the method presented herein compared to these reports result from the bioinformatics and algorithmic development, based on data obtained by a benchmark of hundreds of fingerprints, for dozens of glycovariants, enabling the interpretation of the lectin fingerprints, and providing the user with a list of glycan epitopes upon a mouse-click.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

A lectin array-based methodology for the analysis of protein glycosylation

Rosenfeld¹,

Bangio²,

Gerwig³

et al. 2007

Journal of Biochemical and Biophysical Methods

100

View full text Add to dashboard Cite

Glycosylation is the most versatile and one of the most abundant protein modifications. It has a structural role as well as diverse functional roles in many specific biological functions, including cancer development, viral and bacterial infections, and autoimmunity. The diverse roles of glycosylation in biological processes are rapidly growing areas of research, however, Glycobiology research is limited by the lack of a technology for rapid analysis of glycan composition of glycoproteins. Currently used methods for glycoanalysis are complex, typically requiring high levels of expertise and days to provide answers, and are not readily available to all researcher.We have developed a lectin array-based method, Qproteome™ GlycoArray kits, for rapid analysis of glycosylation profiles of glycoproteins. Glycoanalysis is performed on intact glycoproteins, requiring only 4-6 h for most analysis types. The method, demonstrated in this manuscript by several examples, is based on binding of an intact glycoprotein to the arrayed lectins, resulting in a characteristic fingerprint that is highly sensitive to changes in the protein's glycan composition. The large number of lectins, each with its specific recognition pattern, ensures high sensitivity to changes in the glycosylation pattern. A set of proprietary algorithms automatically interpret the fingerprint signals to provide a comprehensive glycan profile output.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

A lectin array-based methodology for the analysis of protein glycosylation

Rosenfeld¹,

Bangio²,

Gerwig³

et al. 2007

Journal of Biochemical and Biophysical Methods

100

View full text Add to dashboard Cite

show abstract

“…Several labeling techniques exist, including those with detection by optical methods [22,23] and mass spectrometry [24 -26]. One common quantitation strategy using labeling and optical detection involves the use of lectins to bind glycoproteins in complex samples, then detecting different types of glycosylation due to differential binding of the lectins.…”

mentioning

confidence: 99%

“…Because different lectins have different specificities for classes of N-linked glycans, it is possible to use lectins to distinguish between high mannose and complex type glycosylation, for example. Detecting the binding of lectins to glycans, glycopeptides, or glycoproteins is done by either tagging the lectins [22,27] or tagging the analyte [23] with a fluorophore, followed by the monitoring of a change in fluorescence upon binding. If the ultimate quantitative goal is to detect changes in classes of N-linked glycans, optical methods that detect differences in lectin binding are ideal and have very low detection limits.…”

mentioning

confidence: 99%

“…If the ultimate quantitative goal is to detect changes in classes of N-linked glycans, optical methods that detect differences in lectin binding are ideal and have very low detection limits. However, the use of lectin microarrays is incapable of observing subtle glycosylation changes, such as a change in the number of mannoses present on a high mannose glycan [23].…”

mentioning

confidence: 99%

See 1 more Smart Citation

Label-free quantitation: A new glycoproteomics approach

Rebecchi

Wenke

et al. 2009

J. Am. Soc. Mass Spectrom.

View full text Add to dashboard Cite

We demonstrate herein a method for quantifying glycosylation changes on glycoproteins. This novel method uses MS data of characterized glycopeptides to analyze glycosylation profiles, and several quality control tests were done to demonstrate that the method is reproducible, robust, applicable to different types of glycoproteins, and tolerant of instrumental variability during ionization of the analytes. This method is unique in that it is the first label-free quantitative method specifically designed for glycopeptide analysis. It can be used to monitor changes in glycosylation in a glycosylation site-specific manner on a single glycoprotein, or it can be used to quantify glycosylation in a glycoprotein mixture. During mixture analysis, the method can discriminate between changes in glycosylation of a given protein, and changes in the glycoprotein's concentration in the mixture. This method is useful for quantitative analyses in biochemical studies of glycoproteins, where changes in glycosylation composition can be linked to functional differences; it could also be implemented in the pharmaceutical industry, where glycosylation profiles of glycoprotein-based therapeutics must be quantified. Finally, quantification of glycopeptides is an important aspect of glycopeptide-based biomarker discovery, and our quantitative approach could be a valuable asset to this field as well, provided the compositions of the glycopeptides to be quantified are identifiable using other methods. (J Am Soc Mass

show abstract

NMR analysis on the sialic acid‐binding mechanism of an R‐type lectin mutant by natural evolution‐mimicry

et al. 2016

View full text Add to dashboard Cite

A sialic acid-binding lectin (SRC) was created from the C-terminal domain of an R-type N-acetyl lactosamine-binding lectin (EW29Ch) by natural evolution-mimicry. Here, we clarified its sialic acid-binding mechanism using NMR spectroscopy. The NMR analysis showed differences between conformations of the 6 0 -sialyllactose-bound SRC in the solution state and that in the crystal state, and differences between the internal motion of the loop region in subdomain c in SRC and that of the corresponding region in EW29Ch. The NMR analysis thus provided useful information to explain the manner of binding to 6 0 -sialyllactose in solution, which the previous X-ray crystal structure analysis lacked.

show abstract

Development of a Lectin Microarray for the Rapid Analysis of Protein Glycopatterns

Cited by 289 publications

References 33 publications

A lectin array-based methodology for the analysis of protein glycosylation

A lectin array-based methodology for the analysis of protein glycosylation

Label-free quantitation: A new glycoproteomics approach

NMR analysis on the sialic acid‐binding mechanism of an R‐type lectin mutant by natural evolution‐mimicry

Contact Info

Product

Resources

About