Usefulness of gel filtration fraction as potential biomarker for neurocysticercosis in serum: towards a new diagnostic tool

Parasitology ◽  
2016 ◽  
Vol 144 (4) ◽  
pp. 426-435 ◽  
Author(s):  
D. S. NUNES ◽  
H. T. GONZAGA ◽  
V. S. RIBEIRO ◽  
J. P. CUNHA-JÚNIOR ◽  
J. M. COSTA-CRUZ
Keyword(s):  
B Cell ◽  
Calcium Binding ◽  
Binding Protein ◽  
Gel Filtration ◽  
Calcium Binding Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Filtration Fraction ◽  
B Cell Epitopes ◽  
Diagnostic Parameters

SUMMARYThere is an increasing interest in improving neurocysticercosis (NCC) diagnosis through the search of new and alternative antigenic sources, as those obtained from heterologous antigens. The aim of this study was to obtain potential biomarkers for NCC diagnosis after gel filtration chromatography [gel filtration fraction (GFF)] from the total saline extract (SE) from Taenia saginata metacestodes, followed by protein identification and application in immunodiagnostic. SE and GFF proteic profiles were characterized in gel electrophoresis, and diagnostic performance was verified by testing 160 serum samples through enzyme-linked immunosorbent assay and immunoblotting. Sensitivity (Se), specificity (Sp) and other diagnostic parameters were calculated. Polypeptides of interest in the diagnosis of human NCC present at GFF were analysed by mass spectrometry (MS) and B-cell epitopes were predicted. GFF had the best diagnostic parameters: Se 93·3%; Sp 93%; AUC 0·990; LR+ = 13·42 and LR− = 0·07, and proved to be useful reacting with serum samples in immunoblotting. Proteic profile ranged from 64 to 68 kDa and enolase and calcium binding protein calreticulin precursor were identified after MS. The enolase and calcium-binding protein calreticulin precursor showed 18 and 10 predicted B-cell epitopes, respectively. In conclusion we identified important markers in the GFF with high efficiency to diagnose NCC.

Radioimmunoassay Studies of Intestinal Calcium-binding Protein in the Pig. II. The Distribution of Intestinal CaBP in Pig Tissues

1975 ◽  
Vol 53 (6) ◽  
pp. 1135-1140 ◽  
Author(s):  
B. M. Arnold ◽  
M. Kuttner ◽  
D. M. Willis ◽  
A. J. W. Hitchman ◽  
J. E. Harrison ◽  
...  
Keyword(s):  
Small Intestine ◽  
Calcium Binding ◽  
Binding Protein ◽  
Gel Filtration ◽  
Protein S ◽  
Binding Activity ◽  
Calcium Binding Protein ◽  
Distal Small Intestine ◽  
Specific Radioimmunoassay ◽  
Kidney Liver

Using a specific radioimmunoassay for porcine intestinal calcium-binding protein (CaBP), we have measured the concentration of CaBP in the various tissues and organs of normal pigs. Intestinal CaBP was present in highest concentration in the upper small intestine, with lower concentrations in the distal small intestine. Intestinal CaBP was also found, in lower concentrations, in kidney, liver, thyroid, pancreas, and blood. In all other tissues, including parathyroid, bone, skeletal muscle, and brain, CaBP immunoreactivity was undetectable or less than in blood. The elution profile of calcium-binding activity and immunoreactivity from gel filtration analysis of kidney and parathyroid extracts suggest that the calcium-binding protein in the parathyroid gland, and the major calcium-binding protein(s) in the kidney, are chemically and immunochemically different from intestinal CaBP.

scholarly journals Isolation and characterization of grancalcin, a novel 28 kDa EF-hand calcium-binding protein from human neutrophils

1992 ◽  
Vol 286 (2) ◽  
pp. 549-554 ◽  
Author(s):  
C G Teahan ◽  
N F Totty ◽  
A W Segal
Keyword(s):  
Sequence Analysis ◽  
Calcium Binding ◽  
Binding Protein ◽  
Sequence Similarity ◽  
Gel Filtration ◽  
Calcium Binding Protein ◽  
Peptide Sequence ◽  
Human Neutrophils ◽  
Isolation And Characterization ◽  
Ef Hand

A novel 28 kDa protein, which we have named ‘grancalcin’, has been identified in human neutrophils. The protein was isolated from the cytosol and found to be a homodimer, with an apparent molecular mass of 55 kDa on gel filtration. Polyclonal antibodies were raised to the native protein. N-Terminal sequence analysis and tryptic-peptide sequence analysis was performed. The protein exhibits sequence similarity to sorcin, a 24 kDa calcium-binding protein over-expressed in certain multi-drug-resistant cell lines. It appears to be a member of the EF-hand family of calcium-binding proteins. The association of a high proportion of this protein with the membranes and granules in the presence of physiological concentrations of calcium may indicate a role in granule-membrane fusion and degranulation.

scholarly journals Identification and characterization of a calcium-binding protein in the mouse chorioallantoic placenta

1986 ◽  
Vol 233 (1) ◽  
pp. 41-49 ◽  
Author(s):  
R S Tuan ◽  
S T Cavanaugh
Keyword(s):  
Calcium Binding ◽  
Binding Protein ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Calcium Binding Protein ◽  
Chorionic Villi ◽  
Chorioallantoic Placenta ◽  
Identification And Characterization

Mouse chorioallantoic placenta contains a specific calcium-binding protein (MCaBP). A procedure involving gel filtration and ion-exchange chromatography was developed to purify the MCaBP. The MCaBP activity increased as a function of embryonic gestation and was highly specific for Ca2+. The MCaBP is a monomeric protein of Mr 57000, with pI 4.7. Specific antibodies were prepared against the MCaBP and were used to localize the MCaBP to syncytiotrophoblasts of the chorionic villi of mouse chorioallantoic placenta. These properties suggest that the MCaBP may be involved in transplacental calcium transport.

scholarly journals Proteomics detection of S100A6 in tumor tissue interstitial fluid and evaluation of its potential as a biomarker of cholangiocarcinoma

Tumor Biology ◽  
2018 ◽  
Vol 40 (4) ◽  
pp. 101042831876719 ◽  
Author(s):  
Sudarat Onsurathum ◽  
Ornuma Haonon ◽  
Porntip Pinlaor ◽  
Chawalit Pairojkul ◽  
Narong Khuntikeo ◽  
...  
Keyword(s):  
Calcium Binding ◽  
Interstitial Fluid ◽  
Triosephosphate Isomerase ◽  
Biomarker Discovery ◽  
Binding Protein ◽  
Calcium Binding Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Individuals ◽  
S100 Calcium Binding Protein ◽  
Tumor Interstitial Fluid

Tumor interstitial fluid contains tumor-specific proteins that may be useful biomarkers for cancers. In this study, we identified proteins present in cholangiocarcinoma interstitial fluid. Proteins derived from three samples of tumor interstitial fluid and paired samples of adjacent normal interstitial fluid from cholangiocarcinoma patients were subjected to two-dimensional liquid chromatography with tandem mass spectrometry. Candidate proteins were selected based on a greater than twofold change in expression levels between tumor interstitial fluid and normal interstitial fluid. Upregulation of six proteins in tumor interstitial fluid, including S100 calcium binding protein A6 (S100A6), S100 calcium binding protein A9, aldo-keto reductase family 1 member C4, neuropilin-1, 14-3-3 zeta/delta, and triosephosphate isomerase was assessed by western blot and immunohistochemistry. Their potential as markers was evaluated in human cholangiocarcinoma tissue arrays, and in serum using enzyme-linked immunosorbent assay. Expression of S100A6 was higher in tumor interstitial fluid than in normal interstitial fluid and showed the highest positive rate (98.96%) in cholangiocarcinoma tissues. Serum levels of S100A6 did not differ between cholangitis and cholangiocarcinoma patients, but were significantly higher than in healthy individuals ( p < 0.0001). In cholangiocarcinoma cases, S100A6 level was associated with vascular invasion ( p = 0.007) and could distinguish cholangiocarcinoma patients from healthy individuals as effectively as the carbohydrate antigen 19-9. In addition, potential for drug treatment targeting S100A6 and other candidate proteins was also demonstrated using STITCH analysis. In conclusion, proteomics analysis of tumor interstitial fluid could be a new approach for biomarker discovery, and S100A6 is a potential risk marker for screening of cholangiocarcinoma.

scholarly journals Expression of S100 calcium-binding protein A8 in peripheral blood of patients with preeclampsia during pregnancy

2019 ◽  
Vol 17 ◽  
pp. 205873921985852
Author(s):  
Xiaoyun Li ◽  
Fangzheng Cheng ◽  
Guangchao Cao
Keyword(s):  
Calcium Binding ◽  
Binding Protein ◽  
Calcium Binding Protein ◽  
Severe Preeclampsia ◽  
Inflammatory Factors ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Control Groups ◽  
S100 Calcium Binding Protein ◽  
Tnf Α

A total of 30 late onset severe preeclampsia (LS-PE) patients and 30 early onset severe preeclampsia (ES-PE) patients were selected as Experimental group, and 30 normal pregnant were selected as Control group. Expression of S100 calcium-binding protein A8 (S100A8) mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). Enzyme-linked immunosorbent assay (ELISA) was used to detect expression of S100A8 protein and inflammatory factors. Levels of uric acid (UA) and creatinine (CRE) were measured using an automatic biochemical analyzer. Urinary protein (UPRO) content was measured using biuret colorimetry. S100A8 levels were significantly higher in experimental groups than in control groups at mRNA and protein levels ( P < 0.05). Significantly increased contents of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-12, UA, CRE, and UPRO, and decreased level of IL-10 were found in experimental groups than in control groups ( P < 0.05). Compared with patients with ES-PE, significantly higher levels of TNF-α, IL-12, IL-6, UA, CRE, and UPRO, and lower level of IL-10 were found in patients with LS-PE. S100A8 plays pivotal roles in the development of preeclampsia through the interactions with other inflammatory factors.

scholarly journals Antibodies to Both Terminal and Internal B-Cell Epitopes of Francisella tularensisO-Polysaccharide Produced by Patients with Tularemia

2013 ◽  
Vol 21 (2) ◽  
pp. 227-233 ◽  
Author(s):  
Zhaohua Lu ◽  
Hillary M. Perkins ◽  
Jacqueline Sharon
Keyword(s):  
Mouse Model ◽  
B Cell ◽  
Enzyme Linked Immunosorbent Assay ◽  
High Morbidity ◽  
General Strategy ◽  
Serum Samples ◽  
B Cell Epitopes ◽  
Content Type ◽  
Carbohydrate Epitopes ◽  
Secondary Antibodies

ABSTRACTFrancisella tularensis, the Gram-negative bacterium that causes tularemia, is considered a potential bioterrorism threat due to its low infectivity dose and the high morbidity and mortality from respiratory disease. We previously characterized two mouse monoclonal antibodies (MAbs) specific for theO-polysaccharide (O antigen [OAg]) ofF. tularensislipopolysaccharide (LPS): Ab63, which targets a terminal epitope at the nonreducing end of OAg, and Ab52, which targets a repeating internal OAg epitope. These two MAbs were protective in a mouse model of respiratory tularemia. To determine whether these epitope types are also targeted by humans, we tested the ability of each of 18 blood serum samples from 11 tularemia patients to inhibit the binding of Ab63 or Ab52 toF. tularensisLPS in a competition enzyme-linked immunosorbent assay (ELISA). Although all serum samples had Ab63- and Ab52-inhibitory activities, the ratios of Ab63 to Ab52 inhibitory potencies varied 75-fold. However, the variation was only 2.3-fold for sequential serum samples from the same patient, indicating different distributions of terminal- versus internal-binding antibodies in different individuals. Western blot analysis using class-specific anti-human Ig secondary antibodies showed that both terminal- and internal-binding OAg antibodies were of the IgG, IgM, and IgA isotypes. These results support the use of a mouse model to discover protective B-cell epitopes for tularemia vaccines or prophylactic/therapeutic antibodies, and they present a general strategy for interrogating the antibody responses of patients and vaccinees to microbial carbohydrate epitopes that have been characterized in experimental animals.

Calcium Binding Proteins in the Duodenal Mucosa of the Chick, Rat, Pig, and Human

1972 ◽  
Vol 50 (7) ◽  
pp. 758-765 ◽  
Author(s):  
A. J. W. Hitchman ◽  
Joan E. Harrison
Keyword(s):  
Molecular Weight ◽  
Vitamin D ◽  
Calcium Ions ◽  
Calcium Binding ◽  
Binding Protein ◽  
Gel Filtration ◽  
Duodenal Mucosa ◽  
Calcium Binding Proteins ◽  
Calcium Binding Protein ◽  
Electrophoretic Mobilities

A calcium binding protein has been demonstrated in human duodenal mucosa which we believe to be human vitamin D dependent CaBP. Sephadex gel filtration demonstrated that the duodenal mucosa of both the human and pig contained a calcium binding protein with a similar molecular weight to the reported vitamin D dependent rat CaBP (M.W. 12 000–13 000) but dissimilar to chick CaBP (M.W. 24 000–28 000). In addition the rat, pig, and human mucosa contained a high molecular weight calcium binding protein which is probably not vitamin D dependent.A relatively simple procedure, utilizing the tagging of CaBP by 47Ca, has been developed to partially purify normal pig CaBP by Sephadex gel filtration. Further fractionation of the 12 000–13 000 M.W. area, using disc electrophoretic procedures, separated two calcium binding proteins which had similar electrophoretic mobilities and calcium binding formation constants (3.5 and 5.5 × 106, respectively), indicating that both are forms of CaBP but that either one or both have been altered during the procedures. The electrophoretic mobilities of both of these proteins are relatively low in the presence of calcium ions but much greater when calcium ions are removed by chelation with EDTA. This finding should facilitate both the identification and purification of mammalian CaBP.

Sign in / Sign up

Export Citation Format

Share Document