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

Abstract

A 35 kDa glycoprotein whose abundance was previously demonstrated to be enhanced in sera of patients with endometrial adenocarcinoma (n = 12), was isolated from pooled sera of three of the cancer patients using champedak galactose-binding lectin affinity chromatography in the present study. Subjecting it to 2-DE and MS/MS, the glycoprotein was identified as the O-glycosylated fragment of inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4). When compared to control sera (n = 17), expression of the 35 kDa ITIH4 cleavage fragment was demonstrated to be significantly enhanced in sera of patients with breast carcinoma (n = 10), epithelial ovarian carcinoma (n = 10), and germ cell ovarian carcinoma (n = 10) but not in patients with nasopharyngeal carcinoma (n = 13) and osteosarcoma (n = 7). The lectin-based electrophoretic bioanalytical method adopted in the present study may be used to assess the physiological relevance of ITIH4 fragmentation and its correlation with different malignancies, their stages and progression.

SELDI‐TOF as a method for biomarker discovery in the urine of aristolochic‐acid‐treated mice

“…Inter-alpha-trypsin inhibitor heavy chain 4, which was also identified, is a plasma glycoprotein that acts as an acute-phase protein in several species. This protein and its cleavage fragments are also candidate biomarkers associated with different malignant diseases [26,27]. Serum amyloid, transthyretin and inter-alpha-trypsin inhibitor heavy chain 4 were not detected in the urine of AAIItreated mice.…”

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

“…Aside from the use of pre-cast immobilline drystrips pH 3-10 (GE Healthcare Bio-Sciences, Uppsala, Sweden), 2-DE was performed as reported previously [14,15]. Briefly, albumin and IgG-depleted pooled serum samples (30 mg) and bound fractions from the immobilized CMB affinity isolation (30 mg) were subjected to isoelectric focusing using the Multiphor Flatbed electrophoresis system (GE Healthcare Bio-Sciences).…”

Therapeutic plasma proteins – application of proteomics in process optimization, validation, and analysis of the final product

“…Heavy chain H4 of this protein has potential diagnostic value, and it has also been topic of proteomic and glycomic analysis [154]. Weber et al [155] recently described an industrial scale process for production of butyrylcholinesterase.…”