Application of champedak mannose‐binding lectin in the glycoproteomic profiling of serum samples unmasks reduced expression of alpha‐2 macroglobulin and complement factor B in patients with nasopharyngeal carcinoma

The use of lectin affinity chromatography prior to 2‐DE separation forms an alternative method to unmask the expression of targeted glycoproteins of lower abundance in serum samples. Reduced expression of α‐2 macroglobulin (AMG) and complement factor B (CFB) was detected in sera of patients with nasopharyngeal carcinoma (NPC) when pooled serum samples of the patients and those of healthy individuals were subjected to affinity isolation using immobilized champedak mannose‐binding lectin and analyzed by 2‐DE and densitometry. The AMG and CFB spots were not detected in the 2‐DE protein profiles when the same pooled serum samples were subjected to albumin and IgG depletion and neither were they detected when the depleted samples were analyzed by western blotting and lectin detection. Together with other acute‐phase response proteins that were previously reported to be altered in expression in NPC patients, AMG and CFB may serve as useful complementary biomarkers for NPC.

[1]  O. Hashim,et al.  Profiling of serum and tissue high abundance acute-phase proteins of patients with epithelial and germ line ovarian carcinoma , 2008, Proteome Science.

[2]  F. Schena,et al.  Increased serum levels of IgA1-IgG immune complexes and anti-F(ab')2 antibodies in patients with primary IgA nephropathy. , 1989, Clinical and experimental immunology.

[3]  William J Catalona,et al.  Effect of verification bias on screening for prostate cancer by measurement of prostate-specific antigen. , 2003, The New England journal of medicine.

[4]  T. Veenstra,et al.  Characterization of the Low Molecular Weight Human Serum Proteome*S , 2003, Molecular & Cellular Proteomics.

[5]  C. Yip,et al.  Lectin‐based electrophoretic analysis of the expression of the 35 kDa inter‐α‐trypsin inhibitor heavy chain H4 fragment in sera of patients with five different malignancies , 2008, Electrophoresis.

[6]  R. Dernick,et al.  Improved silver staining procedure for fast staining in PhastSystem Development Unit. I. Staining of sodium dodecyl sulfate gels , 1988, Electrophoresis.

[7]  W. Pang,et al.  Can the Acute-Phase Reactant Proteins be Used as Cancer Biomarkers? , 2010, The International journal of biological markers.

[8]  R. Yusof,et al.  Patients with nasopharyngeal carcinoma demonstrate enhanced serum and tissue ceruloplasmin expression. , 2006, The journal of medical investigation : JMI.

[9]  E. Diamandis.  Analysis of serum proteomic patterns for early cancer diagnosis: drawing attention to potential problems. , 2004, Journal of the National Cancer Institute.

[10]  Mary Ann Comunale,et al.  Proteomic analysis of serum associated fucosylated glycoproteins in the development of primary hepatocellular carcinoma. , 2006, Journal of proteome research.

[11]  E. Solomon,et al.  Current status of the molecular characterization of the ovarian cancer antigen CA125 and implications for its use in clinical screening. , 2003, Gynecologic oncology.

[12]  O. Hashim,et al.  THE APPLICATION OF ARTOCARPUS INTEGER SEED LECTIN-M IN THE DETECTION AND ISOLATION OF SELECTIVE HUMAN SERUM ACUTE-PHASE PROTEINS AND IMMUNOGLOBULINS , 2001, Immunological investigations.

[13]  J. Dhainaut,et al.  Hepatic response to sepsis: Interaction between coagulation and inflammatory processes , 2001, Critical care medicine.

[14]  O. Hashim,et al.  Isolation of a mannose-binding and IgE- and IgM-reactive lectin from the seeds of Artocarpus integer. , 1997, Journal of immunological methods.

[15]  O. Hashim,et al.  Expression of high‐abundance proteins in sera of patients with endometrial and cervical cancers: Analysis using 2‐DE with silver staining and lectin detection methods , 2007, Electrophoresis.

[16]  Liang Zhao,et al.  Serum proteome analysis for profiling protein markers associated with carcinogenesis and lymph node metastasis in nasopharyngeal carcinoma , 2008, Clinical & Experimental Metastasis.

[17]  N. Isaacs,et al.  Crystallization and preliminary structural studies of champedak galactose-binding lectin. , 2009, Acta crystallographica. Section F, Structural biology and crystallization communications.

[18]  F. Li,et al.  Proteomics in nasopharyngeal carcinoma , 2008, Cellular and Molecular Life Sciences.

[19]  H. Hayasawa,et al.  A new apoptotic pathway for the complement factor B‐derived fragment Bb , 2000, Journal of cellular physiology.

[20]  K. James.  Interactions between cytokines and α2-macroglobulin , 1990 .

[21]  R D Appel,et al.  Inside SWISS‐2DPAGE database , 1995, Electrophoresis.

[22]  R. Arnold,et al.  Separation of human IgA1 and IgA2 using jacalin-agarose chromatography. , 1987, Journal of immunological methods.

[23]  C. Yip,et al.  Aberrant expression of acute‐phase reactant proteins in sera and breast lesions of patients with malignant and benign breast tumors , 2004, Electrophoresis.

[24]  Wendy Frankel,et al.  Fibrinogen gamma overexpression in pancreatic cancer identified by large-scale proteomic analysis of serum samples. , 2006, Cancer research.

[25]  K. Huse,et al.  Plasma levels of transforming growth factor‐1β and α2‐macroglobulin before and after radical prostatectomy: association to clinicopathological parameters , 2004, The Prostate.

[26]  Iekhsan Othman,et al.  Galactose-binding lectin from the seeds of champedak (Artocarpus integer): sequences of its subunits and interactions with human serum O-glycosylated glycoproteins. , 2002, Biochemical and biophysical research communications.

[27]  R. Wait,et al.  A modified silver staining protocol for visualization of proteins compatible with matrix‐assisted laser desorption/ionization and electrospray ionization‐ mass spectrometry , 2000, Electrophoresis.

[28]  M. Lerner,et al.  Biomarker Identification in Human Pancreatic Cancer Sera , 2008, Pancreas.

[29]  T. Yip,et al.  Identification of Serum Amyloid A Protein As a Potentially Useful Biomarker to Monitor Relapse of Nasopharyngeal Cancer by Serum Proteomic Profiling , 2004, Clinical Cancer Research.