Tyramide signal amplification for antibody-overlay lectin microarray: A strategy to improve the sensitivity of targeted glycan profiling

Danni L. Meany, Laszlo Hackler, Hui Zhang, Daniel W. Chan

Research output: Contribution to journalArticle

27 Scopus citations

Abstract

Antibody-overlay lectin microarray (ALM) has been used for targeted glycan profiling to identify disease-related protein glycoforms. In this context, high sensitivity is desired because it allows for the identification of disease-related glycoforms that are often present at low concentrations. We describe a new tyramide signal amplification (TSA) for the antibody-overlay lectin microarray procedure for sensitive profiling of glycosylation patterns. We demonstrate that TSA increased the sensitivity of the microarray over 100 times for glycan profiling using the model protein prostate specific antigen (PSA). The glycan profile of PSA enriched from LNCAP cells, obtained at a subnanogram level with the aid of TSA, was consistent with the previous reports. We also established the glycan profile of prostate specific membrane antigen (PSMA) using the TSA and ALM. Thus, the TSA for antibody-overlay lectin microarray is a sensitive, rapid, comprehensive, and high-throughput method for targeted glycan profiling and can potentially be used for the identification of disease-related protein glycoforms.

Original languageEnglish (US)
Pages (from-to)1425-1431
Number of pages7
JournalJournal of Proteome Research
Volume10
Issue number3
DOIs
StatePublished - Mar 4 2011

Keywords

  • PSA
  • PSMA
  • antibody-overlay lectin microarray
  • sensitivity
  • targeted glycan profiling
  • tyramide signal amplification

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