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Submitted on May 11, 2006
Accepted on June 7, 2006

Lectin capture strategies combined with mass spectrometry for the discovery of serum glycoprotein biomarkers

Richard R. Drake, E. Ellen Schwegler, Gunjan Malik, Jose I. Diaz, Timothy Block, Anand Mehta, and Oliver John Semmes

Microbiology and Molecular Cell Biology, Eastern Virginia Medical School, Norfolk, Virginia 23507

Corresponding Author: semmesoj{at}evms.edu

The application of mass spectrometry to identify disease biomarkers in clinical fluids like serum using high-throughput protein expression profiling continues to evolve as technology development, clinical study design, and bioinformatics improve. Previous protein expression profiling studies have offered needed insight into issues of technical reproducibility, instrument calibration, sample preparation, study design and supervised bioinformatics data analysis. In this overview, new strategies to increase the utility of protein expression profiling for clinical biomarker assay development are discussed, with an emphasis on utilizing differential lectin-based glycoprotein capture and targeted immuno-based assays. The carbohydrate binding specificities of different lectins offers a biological affinity approach that complements existing mass spectrometer capabilities and retains automated throughput options. Specific examples using serum samples from prostate cancer and hepatocellular carcinoma subjects are provided, along with suggested experimental strategies for integration of lectin based methods into clinical fluid expression profiling strategies. Our example workflow incorporates the necessity of early validation in biomarker discovery using an immuno-based targeted analytical approach that integrates well with upstream discovery technologies.


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