FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W4200381435> ?p ?o ?g. }

• W4200381435 abstract "In this study, we describe an effective protocol for use in a multiplexed high-throughput antibody microarray with glycan binding protein detection that allows for the glycosylation profiling of specific proteins. Glycosylation of proteins is the most prevalent post-translational modification found on proteins, and leads diversified modifications of the physical, chemical, and biological properties of proteins. Because the glycosylation machinery is particularly susceptible to disease progression and malignant transformation, aberrant glycosylation has been recognized as early detection biomarkers for cancer and other diseases. However, current methods to study protein glycosylation typically are too complicated or expensive for use in most normal laboratory or clinical settings and a more practical method to study protein glycosylation is needed. The new protocol described in this study makes use of a chemically blocked antibody microarray with glycan-binding protein (GBP) detection and significantly reduces the time, cost, and lab equipment requirements needed to study protein glycosylation. In this method, multiple immobilized glycoprotein-specific antibodies are printed directly onto the microarray slides and the N-glycans on the antibodies are blocked. The blocked, immobilized glycoprotein-specific antibodies are able to capture and isolate glycoproteins from a complex sample that is applied directly onto the microarray slides. Glycan detection then can be performed by the application of biotinylated lectins and other GBPs to the microarray slide, while binding levels can be determined using Dylight 549-Streptavidin. Through the use of an antibody panel and probing with multiple biotinylated lectins, this method allows for an effective glycosylation profile of the different proteins found in a given human or animal sample to be developed. Introduction Glycosylation of protein, which is the most ubiquitous post-translational modification on proteins, modifies the physical, chemical, and biological properties of a protein, and plays a fundamental role in various biological processes1-6. Because the glycosylation machinery is particularly susceptible to disease progression and malignant transformation, aberrant glycosylation has been recognized as early detection biomarkers for cancer and other diseases 7-12. In fact, most current cancer biomarkers, such as the L3 fraction of α-1 fetoprotein (AFP) for hepatocellular carcinoma 13-15, and CA199 for pancreatic cancer 16, 17 are all aberrant glycan moieties on glycoproteins. However, methods to study protein glycosylation have been complicated, and not suitable for routine laboratory and clinical settings. Chen et al. has recently invented a chemically blocked antibody microarray with a glycan-binding protein (GBP) detection method for high-throughput and multiplexed profile glycosylation of native glycoproteins in a complex sample 18. In this affinity based microarray method, multiple immobilized glycoprotein-specific antibodies capture and isolate glycoproteins from the complex mixture directly on the microarray slide, and the glycans on each individual captured protein are measured by GBPs. Because all normal antibodies contain N-glycans which could be recognized by most GBPs, the critical step of this method is to chemically block the glycans on the antibodies from binding to GBP. In the procedure, the cis-diol groups of the glycans on the antibodies were first oxidized to aldehyde groups by using NaIO4 in sodium acetate buffer avoiding light. The aldehyde groups were then conjugated to the hydrazide group of a cross-linker, 4-(4-N-MaleimidoPhenyl)butyric acid Hydrazide HCl (MPBH), followed by the conjugation of a dipeptide, Cys-Gly, to the maleimide group of the MPBH. Thus, the cis-diol groups on glycans of antibodies were converted into bulky none hydroxyl groups, which hindered the lectins and other GBPs bindings to the capture antibodies. This blocking procedure makes the GBPs and lectins bind only to the glycans of captured proteins. After this chemically blocking, serum samples were incubated with the antibody microarray, followed by the glycans detection by using different biotinylated lectins and GBPs, and visualized with Cy3-streptavidin. The parallel use of an antibody panel and multiple lectin probing provides discrete glycosylation profiles of multiple proteins in a given sample 18-20. This method has been used successfully in multiple different labs 1, 7, 13, 19-31. However, stability of MPBH and Cys-Gly, complicated and extended procedure in this method affect the reproducibility, effectiveness and efficiency of the method. In this new protocol, we replaced both MPBH and Cys-Gly with one much more stable reagent glutamic acid hydrazide (Glu-hydrazide), which significantly improved the reproducibility of the method, simplified and shorten the whole procedure so that the it can be completed within one working day. In this new protocol, we describe the detailed procedure of the protocol which can be readily adopted by normal labs for routine protein glycosylation study and techniques which are necessary to obtain reproducible and repeatable results." @default.
• W4200381435 created "2021-12-31" @default.
• W4200381435 creator A5003997838 @default.
• W4200381435 creator A5041693287 @default.
• W4200381435 creator A5046744833 @default.
• W4200381435 creator A5057639560 @default.
• W4200381435 creator A5075024050 @default.
• W4200381435 creator A5079636501 @default.
• W4200381435 creator A5086530801 @default.
• W4200381435 date "2012-05-04" @default.
• W4200381435 modified "2023-10-14" @default.
• W4200381435 title "Chemically-blocked Antibody Microarray for Multiplexed High-throughput Profiling of Specific Protein Glycosylation in Complex Samples" @default.
• W4200381435 doi "https://doi.org/10.3791/3791-v" @default.
• W4200381435 hasPublicationYear "2012" @default.
• W4200381435 type Work @default.
• W4200381435 citedByCount "0" @default.
• W4200381435 crossrefType "journal-article" @default.
• W4200381435 hasAuthorship W4200381435A5003997838 @default.
• W4200381435 hasAuthorship W4200381435A5041693287 @default.
• W4200381435 hasAuthorship W4200381435A5046744833 @default.
• W4200381435 hasAuthorship W4200381435A5057639560 @default.
• W4200381435 hasAuthorship W4200381435A5075024050 @default.
• W4200381435 hasAuthorship W4200381435A5079636501 @default.
• W4200381435 hasAuthorship W4200381435A5086530801 @default.
• W4200381435 hasBestOaLocation W42003814352 @default.
• W4200381435 hasConcept C104317684 @default.
• W4200381435 hasConcept C108625454 @default.
• W4200381435 hasConcept C150194340 @default.
• W4200381435 hasConcept C153911025 @default.
• W4200381435 hasConcept C159654299 @default.
• W4200381435 hasConcept C171196145 @default.
• W4200381435 hasConcept C185592680 @default.
• W4200381435 hasConcept C186836561 @default.
• W4200381435 hasConcept C195814341 @default.
• W4200381435 hasConcept C203014093 @default.
• W4200381435 hasConcept C204628709 @default.
• W4200381435 hasConcept C206212055 @default.
• W4200381435 hasConcept C2777313579 @default.
• W4200381435 hasConcept C2910703447 @default.
• W4200381435 hasConcept C46111723 @default.
• W4200381435 hasConcept C55493867 @default.
• W4200381435 hasConcept C70721500 @default.
• W4200381435 hasConcept C71829478 @default.
• W4200381435 hasConcept C86803240 @default.
• W4200381435 hasConcept C95371953 @default.
• W4200381435 hasConceptScore W4200381435C104317684 @default.
• W4200381435 hasConceptScore W4200381435C108625454 @default.
• W4200381435 hasConceptScore W4200381435C150194340 @default.
• W4200381435 hasConceptScore W4200381435C153911025 @default.
• W4200381435 hasConceptScore W4200381435C159654299 @default.
• W4200381435 hasConceptScore W4200381435C171196145 @default.
• W4200381435 hasConceptScore W4200381435C185592680 @default.
• W4200381435 hasConceptScore W4200381435C186836561 @default.
• W4200381435 hasConceptScore W4200381435C195814341 @default.
• W4200381435 hasConceptScore W4200381435C203014093 @default.
• W4200381435 hasConceptScore W4200381435C204628709 @default.
• W4200381435 hasConceptScore W4200381435C206212055 @default.
• W4200381435 hasConceptScore W4200381435C2777313579 @default.
• W4200381435 hasConceptScore W4200381435C2910703447 @default.
• W4200381435 hasConceptScore W4200381435C46111723 @default.
• W4200381435 hasConceptScore W4200381435C55493867 @default.
• W4200381435 hasConceptScore W4200381435C70721500 @default.
• W4200381435 hasConceptScore W4200381435C71829478 @default.
• W4200381435 hasConceptScore W4200381435C86803240 @default.
• W4200381435 hasConceptScore W4200381435C95371953 @default.
• W4200381435 hasIssue "63" @default.
• W4200381435 hasLocation W42003814351 @default.
• W4200381435 hasLocation W42003814352 @default.
• W4200381435 hasOpenAccess W4200381435 @default.
• W4200381435 hasPrimaryLocation W42003814351 @default.
• W4200381435 hasRelatedWork W1488329278 @default.
• W4200381435 hasRelatedWork W1933311620 @default.
• W4200381435 hasRelatedWork W1967095214 @default.
• W4200381435 hasRelatedWork W1970595852 @default.
• W4200381435 hasRelatedWork W2014413829 @default.
• W4200381435 hasRelatedWork W2022725835 @default.
• W4200381435 hasRelatedWork W2094928744 @default.
• W4200381435 hasRelatedWork W2346668946 @default.
• W4200381435 hasRelatedWork W2496175704 @default.
• W4200381435 hasRelatedWork W4283028851 @default.
• W4200381435 isParatext "false" @default.
• W4200381435 isRetracted "false" @default.
• W4200381435 workType "article" @default.