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HPG-ALD Polymers for Proteomics


Project TitleHPG-ALD Polymers for Proteomics
Track Code08-038
Short Description

Novel high molecular weight dendritic hyperbranched polyglycerol-aldehydes (HPG-ALD) are available for purchase. HPG-ALDs can be used for selective peptide separation for mass spectrometry based proteomic analysis, including the terminal amine isotopic labeling of substrates (TAILS) protocol.

Tagsbiomaterials, health & life sciences, r&d discovery, science & technology
Posted DateApr 7, 2010 11:34 AM


Proteomes are tissue specific and dynamically change depending on the physiological and pathological state of the tissues. Mass spectrometry (MS) based functional proteomics has emerged as a powerful tool in quantifying and analyzing proteome modifications.


This technology is the development of a new enrichment method and reagent for proteome analysis, possessing high selectivity as well as high efficiency, while being extremely economical.

• HPG-ALD: Researchers at The University of British Columbia have characterized and developed a novel method to synthesize a novel class of highly efficient, highly functional, high molecular weight (~565 KDa) dendritic polymer supports that contain specific functionalities (~3200/molecule).

• TAILS: Terminal amine isotopic labeling of substrates is a 3-d, three-step quantitative proteomics approach, using HPG-ALD for labeling and isolating N-terminal peptides before and after exposure to a protease of interest. This polymer, when used for TAILS, is also applicable for analysis of proteolysis in cell culture, animal models of disease, and human tissue.

• These water soluble dendritic polyglycerol aldehyde polymers contain both cleavable and non-cleavable groups for the selective separation and quantification of tryptic peptides in proteomic analysis.

• The peptides being analyzed are highly reactive to the functional moiety on the polymer.  

Applications and Advantages


• These highly efficient HPG-ALDs are a versatile tool in proteomics for the identification of enzyme targets and substrates. 

• The polymers are chemically versatile and can incorporate other chemical functionalities for various applications.

• Being water soluble, these new polymer supports are highly efficient in capturing peptides and proteins and require no solid phase reagents; many times better than currently available supports.

• The peptide binding capacity and capture efficiency is a three-fold improvement on current techniques.

• The polymers have very low non-specific binding to peptides.

State of Development

The efficacy of the new polymers has been successfully tested and is shown to discriminate between various peptides from a mixture for both synthetic peptides and a tryptic digest of samples to simplify MS based peptide analysis.

Reference: Kleifeld, O. et al. 2010. Isotopic Labeling of Terminal Amines in Complex Samples Identifies Protein N-termini and Protease Cleavage Products. Nature Biotechnology 28, 281-288. DOI: 10.1038/nbt.1611.


Principal Inventors:

Dr. Christopher Overall and Dr. Jayachandran Kizhakkedathu


Novel high molecular weight dendritic hyperbranched polyglycerol-aldehydes (HPG-ALD; 90 kDa with 516 aldehyde groups) are available for purchase under OFFERINGS.


Klein, T., Fung, S.Y., Renner, F., Blank, M.A., Dufour, A., Kang, S., Bolger-Munro, M., Scurll, J.M., Priatel, J.J., Schweigler, P., Melkko, S., Gold, M.R., Viner, R.I., Regnier, C.H., Turvey, S.E. and Overall, C.M. 2015. The Paracaspase MALT1 Cleaves HOIL1 Reducing Linear Ubiquitination By LUBAC to Dampen Lymphocyte NF-κB Signalling. Nature Communications 6, 8777, 1-17. DOI: 10.1038/ncomms9777.

Huesgen, P.F., Lange, P.F., Rogers, L.D., Solis, N., Eckhard, U., Kleifeld, O., Goulas, T., Gomis-Ruth, F.X., Overall, C.M. 2015. LysargiNase Mirrors Trypsin for Protein C-Terminal and Methylation-Site Identification. Nature Methods 12, 55-58. DOI: 10.1038/nmeth.3177.

Fortelny, N., Yang, S., Pavlidis, P., Lange, P.F. and Overall, C.M. 2015. Proteome TopFIND 3.0 with TopFINDer and PathFINDer: Database and Analysis Tools for the Association of Protein Termini to Pre-and Post-translational Events. Nucleic Acid Research 43, (Database Issue) D290-297. DOI: 10.1093/nar/gku1012.

Fortelny, N., Pavlidis, P. and Overall, C.M. 2015. The Path of No Return - Truncated Protein N-termini and Current Ignorance of their Genesis. Proteomics 15, 14, 2547-2552. DOI: 10.1002/pmic.201500043.

Marino, G., Eckhard, U. and Overall, C.M. 2015 Protein Termini and their Modification Revealed by Positional Proteomics. ACS Chemical Biology 10, 8, 1754-1764. DOI: 10.1021/acschembio.5b00189.

Eckhard, U., Marino G., Abbey, S.R., Tharmarajah, G., Matthew, I. and Overall, C.M. 2015. The Human Dental Pulp Proteome and N-terminome: Levering the Unexplored Potential of Semitryptic Peptides Enriched by TAILS to Identifying Missing Proteins in the Human Proteome Project in Underexplored Tissues. Journal of Proteome Research 14, 9, 3568-3582. DOI: 10.021/acs.jproteome.5b00579.

auf dem Keller, U., Prudova, A., Eckhard, U., Fingleton, B. and Overall, C.M. 2013. Systems-Level Analysis of Proteolytic Events in Increased Vascular Permeability and Complement Activation in Skin Inflammation. Science Signaling 6, 258, rs2, 1-15: DOI: 10.1126/scisignal.2003512.

Lange, P.F. and Overall, C.M. 2013. The TAILS of Proteins: When Protein Termini Tell Tales of Proteolysis and Function. Current Opinion in Chemical Biology 17, 1, 73-82. DOI: 10.1016/j.cbpa.2012.11.025.

Lange, P., Huesgen, P. and Overall, C.M. 2012. TopFIND 2.0—Linking Protein Termini with Proteolytic Processing and Modifications Altering Protein Function. Nucleic Acids Research 40, (Database Issue), D351-361. DOI:

auf dem Keller, U. and Overall, C.M. 2012. CLIPPER—An Add-on to the Trans-Proteomic Pipeline for the Automated Analysis of TAILS N-terminomics Data. Biological Chemistry 393, 12, 1477-1483. DOI: 10.1515/hsz-2012-0269.

Lange, P.F. and Overall, C.M. 2011. TopFIND, a Knowledgebase Linking Protein Termini with Function. Nature Methods 8, 703-704. DOI: 10.1038/nmeth.1669.

Schilling, O., Huesgen, P.F., Barré, O., auf dem Keller, U. and Overall, C.M. 2011. Characterization of the Prime and Non-prime Active Site Specificities of Proteases by Proteome-derived Peptide Libraries and Tandem Mass Spectrometry. Nature Protocols 6, 111-120. DOI: 10.1038/nprot.2010.178.

Doucet, A. and Overall, C.M. 2011. Broad Coverage Identification of Multiple Proteolytic Cleavage Site Sequences in Complex High Molecular Weight Proteins Using Quantitative Proteomics as a Complement to Edman Sequencing. Molecular & Cellular Proteomics 10, 5, M110.003533. DOI: 10.1074/mcp.M110.003533.

Doucet, A., Kleifeld, O., Kizhakkedathu, J.N. and Overall, C.M. 2011. Identification of Proteolytic Products and Natural Protein N-termini by Terminal Amine Isotopic Labeling of Substrates (TAILS). In: Gel-Free Proteomics: Methods and Protocols, edited by Gevaert, K. and Vandekerckhove, J. Methods in Molecular Biology 753, 273-287. Springer Science + Business Media.

Schilling, O., Huesgen, P.F., Barré, O. and OveralI, C.M. 2011. Identification and Relative Quantification of Native and Proteolytically Generated Protein C-termini from Complex Proteomes: C-terminome Analysis. Network Biology: Methods and Applications, edited by Cagney, G. and Emili, A. Methods in Molecular Biology 781, 59-69. Springer Science + Business Media.

Schilling, O., Barré, O., Huesgen, P.F. and Overall, C.M. 2010. Proteome-wide Analysis of Protein Carboxy Termini: C Terminomics. Nature Methods 7, 508-511. DOI: 10.1038/nmeth.1467Featured in C&EN (Chemical and Engineering News).

Kleifeld, O., Doucet, A., auf dem Keller, U., Prudova, A., Schilling, O., Kainthan, R.K., Starr, A.E., Foster, L.J., Kizhakkedathu, J.N. and Overall, C.M. 2010. Isotopic Labeling of Terminal Amines in Complex Samples Identifies Protein N-termini and Protease Cleavage Products. Nature Biotechnology 28, 281-288. DOI: 10.1038/nbt.1611.

Kleifeld, O., Doucet, A., Prudova, A., auf dem Keller, U., Gioia, M., Kizhakkedathu, J.N., and Overall, C.M. 2011. Identifying and Quantifying Proteolytic Events and the Natural N-terminome by Terminal Amine Isotopic Labeling of Substrates. Nature Protocols 6, 10, 1578-1611. DOI: 10.1038/nprot.2011.382.

Prudova, A., auf dem Keller, U., Butler, G.S. and Overall, C.M. 2010. Multiplex N-terminome Analysis of MMP-2 and MMP-9 Substrate Degradomes by iTRAQ-TAILS Quantitative Proteomics. Molecular Cellular Proteomics 9, 5, 894-911. DOI: 10.1074/mcp.M000050-MCP201.

Lange, P.F., Huesgen, P.F., Nguyen, K. and Overall, C.M. 2014. Annotating N Termini for the Human Proteome Project: N Termini and Nα-Acetylation Status Differentiate Stable Cleaved Protein Species from Degradation Remnants in the Human Erythrocyte Proteome. Journal of Proteome Research 13, 4, 2028-2044. DOI:

Bellac, C.L., Dufour, A., Krisinger, M.J., Loonchanta, A., Starr, A.E., auf dem Keller, U., Lange, P.F., Goebeler, V., Kappelhoff, R., Butler, G.S., Burtnick, L.D., Conway, E.M., Roberts, C.R. and Overall, C.M. 2014. Macrophage Matrix Metalloproteinase-12 Dampens Inflammation and Neutrophil Influx in Arthritis. Cell Reports 9, 2, 618-632. DOI:


File Name Description
TAILS Protocol None Download


Name Price
HPG-ALD Polymer - 20mg aliquots NEW lower price - Academic 600.00 USD to 350.00 USD Buy
HPG-ALD Polymer - 20mg aliquots NEW lower price - Commercial and Core Facilities 1,100.00 USD to 850.00 USD Buy