One-Pot Two-Nanoprobe Assay Uncovers Targeted Glycoprotein Biosignature

被引:27
作者
dela Rosa, Mira Anne C. [1 ,3 ,4 ]
Chen, Wei-Chun [4 ,5 ]
Chen, Yi-Ju [4 ]
Obena, Rofeamor P. [4 ]
Chang, Chih-Hsiang [4 ]
Capangpangan, Rey Y. [4 ,6 ]
Su, Tung-Hung [2 ]
Chen, Chi-Ling [2 ]
Chen, Pei-Jer [2 ]
Chen, Yu-Ju [1 ,4 ]
机构
[1] Natl Taiwan Univ, Coll Med, Dept Chem, Taipei, Taiwan
[2] Natl Taiwan Univ, Coll Med, Grad Inst Clin Med, Taipei, Taiwan
[3] Acad Sinica, Taiwan Int Grad Program, Nano Sci & Technol Program, Taipei, Taiwan
[4] Acad Sinica, Inst Chem, Taipei, Taiwan
[5] Natl Taiwan Normal Univ, Dept Chem, Taipei, Taiwan
[6] Caraga State Univ, Butuan City, Philippines
关键词
SERUM ALPHA-FETOPROTEIN; HEPATOCELLULAR-CARCINOMA; MASS-SPECTROMETRY; EARLY-DIAGNOSIS; SUGAR CHAIN; PROTEIN; GLYCOSYLATION; BIOMARKERS; PROTEOMICS; LIVER;
D O I
10.1021/acs.analchem.6b04396
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
We report a one-pot two-nanoprobe approach coupled to mass spectrometry for simultaneous quantification and post-translational modification (PTM) profiling of targeted protein in biofluid. Using N-glycoprotein as model, the assay employs two nanoprobes, antibody-conjugated SiO2 nanoparticles and lectin-conjugated magnetic Fe3O4 nano particles, to achieve target glycoprotein isolation from biofluid and subsequent glycopeptide enrichment in-a single tube. As demonstrated on a-fetoprotein (AFP), a serum biomarker for hepatocellular carcinoma (HCC), the assay has high purification specificity (20 glycopeptides);with 2-fold and 10-fold superior total glycopeptide intensity compared to non one -pot method (9 glycopeptides) or without enrichment (6, glycopeptides), respectively. By multiple reaction monitoring mass spectrometry (MRM-MS) analysis of the nonglycopeptides, the assay can quantify low abundant AFP expression (0.5 ng) with good correlation with conventional ELISA method (Pearson's r = 0.987). Furthermore, we present the first study revealing AFP glycopeptide signatures of individual HCC patients, comprised of 23 heterogeneous glycoforms of bi-and triantennary, core and terminal fucosylation, and mono-to trisialylation. In addition to 12 novel AFP glycoforms, our quantification result uncovers five abundant glycoforms in HCC, including 3 core-fucosylated (CF) forms. These identified CF forms may be 'evaluated in future studies as potential targets'in a glycopeptide biomarker panel to further improve accuracy of conventional AFP-L3 tests. Through this one-pot assay, a comprehensive target protein profile comprised of protein expression-and glycosylation pattern was achieved in simple protocol 'with high sensitivity, reduced analysis time, and minute starting material. This assay can be extended to other PTM biosignatures by conjugation of other affinity ligands on the nanoprobe.
引用
收藏
页码:3973 / 3980
页数:8
相关论文
共 50 条
[1]   N-linked Glycan Structures and Their Expressions Change in the Blood Sera of Ovarian Cancer Patients [J].
Alley, William R., Jr. ;
Vasseur, Jacqueline A. ;
Goetz, John A. ;
Syoboda, Martin ;
Mann, Benjamin F. ;
Matei, Daniela E. ;
Menning, Nancy ;
Hussein, Ahmed ;
Mechref, Yehia ;
Novotny, Milos V. .
JOURNAL OF PROTEOME RESEARCH, 2012, 11 (04) :2282-2300
[2]   CARBOHYDRATE STRUCTURES OF HUMAN ALPHA-FETOPROTEIN OF PATIENTS WITH HEPATOCELLULAR-CARCINOMA - PRESENCE OF FUCOSYLATED AND NONFUCOSYLATED TRIANTENNARY GLYCANS [J].
AOYAGI, Y ;
SUZUKI, Y ;
IGARASHI, K ;
SAITOH, A ;
OGURO, M ;
YOKOTA, T ;
MORI, S ;
SUDA, T ;
ISEMURA, M ;
ASAKURA, H .
BRITISH JOURNAL OF CANCER, 1993, 67 (03) :486-492
[3]   UniProt: a hub for protein information [J].
Bateman, Alex ;
Martin, Maria Jesus ;
O'Donovan, Claire ;
Magrane, Michele ;
Apweiler, Rolf ;
Alpi, Emanuele ;
Antunes, Ricardo ;
Arganiska, Joanna ;
Bely, Benoit ;
Bingley, Mark ;
Bonilla, Carlos ;
Britto, Ramona ;
Bursteinas, Borisas ;
Chavali, Gayatri ;
Cibrian-Uhalte, Elena ;
Da Silva, Alan ;
De Giorgi, Maurizio ;
Dogan, Tunca ;
Fazzini, Francesco ;
Gane, Paul ;
Cas-tro, Leyla Garcia ;
Garmiri, Penelope ;
Hatton-Ellis, Emma ;
Hieta, Reija ;
Huntley, Rachael ;
Legge, Duncan ;
Liu, Wudong ;
Luo, Jie ;
MacDougall, Alistair ;
Mutowo, Prudence ;
Nightin-gale, Andrew ;
Orchard, Sandra ;
Pichler, Klemens ;
Poggioli, Diego ;
Pundir, Sangya ;
Pureza, Luis ;
Qi, Guoying ;
Rosanoff, Steven ;
Saidi, Rabie ;
Sawford, Tony ;
Shypitsyna, Aleksandra ;
Turner, Edward ;
Volynkin, Vladimir ;
Wardell, Tony ;
Watkins, Xavier ;
Zellner, Hermann ;
Cowley, Andrew ;
Figueira, Luis ;
Li, Weizhong ;
McWilliam, Hamish .
NUCLEIC ACIDS RESEARCH, 2015, 43 (D1) :D204-D212
[4]   Quantification of Fucosylated Hemopexin and Complement Factor H in Plasma of Patients with Liver Disease [J].
Benicky, Julius ;
Sanda, Miloslav ;
Pompach, Petr ;
Wu, Jing ;
Goldman, Radoslav .
ANALYTICAL CHEMISTRY, 2014, 86 (21) :10716-10723
[5]  
Bern Marshall, 2012, Curr Protoc Bioinformatics, VChapter 13, DOI 10.1002/0471250953.bi1320s40
[6]   Alteration of protein glycosylation in liver diseases [J].
Blomme, Bram ;
Van Steenkiste, Christophe ;
Callewaert, Nico ;
Van Vlierberghe, Hans .
JOURNAL OF HEPATOLOGY, 2009, 50 (03) :592-603
[7]   Oncogenic kinase signalling [J].
Blume-Jensen, P ;
Hunter, T .
NATURE, 2001, 411 (6835) :355-365
[8]   UniCarbKB: building a knowledge platform for glycoproteomics [J].
Campbell, Matthew P. ;
Peterson, Robyn ;
Mariethoz, Julien ;
Gasteiger, Elisabeth ;
Akune, Yukie ;
Aoki-Kinoshita, Kiyoko F. ;
Lisacek, Frederique ;
Packer, Nicolle H. .
NUCLEIC ACIDS RESEARCH, 2014, 42 (D1) :D215-D221
[9]   Glycoprotein Disease Markers and Single Protein-omics [J].
Chandler, Kevin ;
Goldman, Radoslav .
MOLECULAR & CELLULAR PROTEOMICS, 2013, 12 (04) :836-845
[10]  
Chandramouli Kondethimmanahalli, 2009, Hum Genomics Proteomics, V2009, DOI 10.4061/2009/239204