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Journal of Immunoassay & Immunochemistry

Madan Kumar Paudel, Seiichi Sakamoto, Le Van Huy, Hiroyuki Tanaka, Miyamoto Tomofumi, Takano Akihito, Satoshi Morimoto
Wogonin 7-O-β-D-glucuronide (Wgn) is a bioactive flavone present in the dried root of Scutellaria baicalensis Georgi. To generate a monoclonal antibody (MAb) against Wgn, BALB/c mice injected with Wgn-bovine serum albumin yielded splenocytes that we fused with SP2/0 myeloma cells using the polyethylene glycol method. We obtained a hybridoma designated 315A that produced a MAb reactive to Wgn. The anti-Wgn MAb 315A was applied to an indirect competitive enzyme-linked immunosorbent assay (icELISA) to quantify Wgn...
December 27, 2016: Journal of Immunoassay & Immunochemistry
Asiabanha Rezaee Majid, Rasaee Mohammad Javad, Paknejad Malihe, Mohammadnejad Javad
The Human Cardiac Troponin I (hcTnI) is a 210 amino acid protein, 23 kDa in molecular weight. This biomarker is commonly used to diagnose myocardial infarction, micro injury, and acute coronary syndrome (ACS) in patients referring to emergency departments. The American Heart Association (AHA) and European Society of Cardiology (ESC) proposed troponin I as the gold biomarker for early detection of heart attack, especially in myocardial infarction (MI). Therefore, developing monoclonal antibodies against this biomarker could help in for early detection of heart attack...
December 27, 2016: Journal of Immunoassay & Immunochemistry
V O Oyebanji, B O Emikpe, Omolade A Oladele, M O Odeniyi, A Salami, O I Osowole, O B Kasali, O A Akinboade
Immune response of challenged chickens following previous vaccinations with Newcastle disease vaccine using gums from Cedrela odorata and Khaya senegalensis as delivery agent were evaluated. Two hundred and fifty-two (252) one-day old chickens were divided into vaccine-gum oral (GVOR), vaccine-gum ocular (GVOC), vaccine oral (VOR), vaccine ocular (VOC), gum oral (GOR), gum ocular (GOC), No-gum-no-vaccine/challenged (NGNV/C) & No-gum-no-vaccine/unchallenged (NGNV/U) groups. They were vaccinated at days 21 & 42 and challenged at day 84...
December 23, 2016: Journal of Immunoassay & Immunochemistry
Justina O Tam, Helena de Puig, Chun-Wan Yen, Irene Bosch, Jose Gómez-Márquez, Charles Clavet, Kimberly Hamad-Schifferli, Lee Gehrke
Point-of-care (POC) diagnostics such as lateral flow and dipstick immunoassays use gold nanoparticle (NP)-antibody conjugates for visual readout. We investigated the effects of NP conjugation, surface chemistries, and antibody immobilization methods on dipstick performance. We compared orientational, covalent conjugation, electrostatic adsorption, and a commercial conjugation kit for dipstick assays to detect dengue virus NS1 protein. Assay performance depended significantly on their conjugate properties. We also tested arrangements of multiple test lines within strips...
December 16, 2016: Journal of Immunoassay & Immunochemistry
Ahad Khalilnezhad, Elham Mahmoudian, Nariman Mosaffa, Jaleh Mohsenifar, Davar Amani
BACKGROUND: We hypothesized that Tumor cell lysate (TCL) prepared from spontaneous mouse mammary tumor (SMMT) may elicit IgG production by spleen mononuclear cells (SMCs) in ex vivo. METHODS: The SMCs from healthy mice (HM, n = 6) and four-week SMMT-bearing mice (TBM, n = 6) was cultured in presence of TCL and mitogen for 42 hr at 37°C, separately. Serum and SMCs culture supernatant levels of IFNγ, IL-4, and total IgG were measured using special ELISA kits. RESULTS: Serum IgG level of TBM was significantly higher than that of HM group (P = 0...
December 8, 2016: Journal of Immunoassay & Immunochemistry
Shunsuke Fujii, Osamu Morinaga, Takuhiro Uto, Shuichi Nomura, Yukihiro Shoyama
Immunoassay systems using monoclonal antibodies (mAbs) are one of the most useful techniques in the analytical, biochemical, and clinical fields. In this study, a combination enzyme-linked immunosorbent assay (ELISA) using both anti-glycyrrhizin and anti-liquiritin mAbs (anti-GL/Liq mixture mAbs) was developed for quality control of licorice and its products. The combination ELISA demonstrated high sensitivity, reproducibility, and specificity for the total content of GL and Liq by a single assay. The developed ELISA was effective and useful as the first screening method in the selection of high-quality licorice from the Glycyrrhiza species and in confirming the quality of licorice-containing Kampo medicines...
November 28, 2016: Journal of Immunoassay & Immunochemistry
Susanne Pribbenow, Tulsidas G Shrivastav, Martin Dehnhard
Enzyme-immunoassays (EIA) that detect fecal testosterone metabolites (fTM) are powerful tools to monitor gonadal activity non-invasively. However, a challenge with testosterone EIAs might be their potential for cross-reactivities with structurally similar glucocorticoid metabolites. Therefore, we aimed to verify the capability of four different testosterone EIAs to monitor fTM without reflecting changes in adrenocortical activity in spotted hyenas by analyzing fecal samples following testosterone and ACTH challenge tests...
November 17, 2016: Journal of Immunoassay & Immunochemistry
Adelina Rosca, Gabriela Anton, Luminita Ene, Iulia Iancu, Aura Temereanca, Cristian L Achim, Simona M Ruta
This study aimed to investigate the influence of antiretroviral therapy on methylation markers, in a group of HIV infected, heavily treated patients. Immune and molecular methods were used to investigate potential changes in methylation profile in DNA isolated from peripheral blood mononuclear cells collected from antiretroviral-experienced HIV infected patients and healthy controls. The percentage of 5-methylcytosine was inversely correlated with proviral DNA and active replication while DNMT1 (p = 0.01) and DNMT3A (p = 0...
November 17, 2016: Journal of Immunoassay & Immunochemistry
Bharti Jain, J Kumarasamy, C Gholve, Savita Kulkarni, M G R Rajan
We describe the development and validation of multianalyte immunoassays (MAIA) for three analytes, viz., thyroxine (T4), thyroid stimulating hormone (TSH), and thyroglobulin (Tg) essential for assessment of thyroid function but having widely varying molecular weights. Using polycarbonate (PC) track-etched membranes (TEM) as an immobilization support and (125)I as the tracer, both competitive assay for T4 and non-competitive assay for TSH and Tg were performed on the same TEM. MAIA was found to be highly sensitive and precise with clinically useful working range and correlated very well with individual analyte immunoassays...
November 1, 2016: Journal of Immunoassay & Immunochemistry
Ana Rodríguez-Rodríguez, Juan José Egea-Guerrero, Ángel Vilches-Arenas, Manuel Jesús Quintanilla-Vázquez, Francisco Murillo-Cabezas, M Ángeles Muñoz-Sánchez
Changes in Urotensin-II (U-II) concentration, a potent vasoconstrictor peptide, have been detected in various pathologies, but it has been impossible to define a normality range. We aimed to analyze the concordance and interchangeability between two enzyme immunoassay methods developed by Phoenix Pharmaceuticals, Inc. to measure U-II plasma concentration in rats: ELISA and fluorescent EIA. Assays resulted positively correlated (r = 0.850; p < 0.01). There was a significant difference between assays values (p < 0...
November 1, 2016: Journal of Immunoassay & Immunochemistry
Presentini Rivo
A new method has been developed to prepare horseradish peroxidase (HRP) conjugates using bis(sulfosuccinimidyl)suberate (BS3) as cross-linking reagent in a two-step procedure. The enzyme is reacted with BS3, introducing active ester molecules into the enzyme itself, and it is then used directly to label amino-containing compounds without further treatment. The proteins involved in the conjugation undergo minimal modifications. The reaction conditions are evaluated, as well as studies on the conservation of the biological activities of the conjugated proteins and the stability of the conjugates in time...
October 31, 2016: Journal of Immunoassay & Immunochemistry
Maysam Mard-Soltani, Mohamad Javad Rasaee, AbdolKarim Sheikhi, Mehdi Hedayati
Designing novel antigens to rise specific antibodies for Thyroid Stimulating Hormone (TSH) detection is of great significance. A novel fusion protein consisting of the C termini sequence of TSH beta subunit and a fusion sequence was designed and produced for rabbit immunization. Thereafter, the produced antibodies were purified and characterized for TSH detection. Our results indicate that the produced antibody is capable of sensitive and specific detection of TSH with low cross reactivity. This study underscores the applicability of designed fusion protein for specific and sensitive polyclonal antibody production and the importance of selecting an amenable region of the TSH for immunization...
October 27, 2016: Journal of Immunoassay & Immunochemistry
Awoyelu E Hilda, Oloke Julius Kola, Oladipo E Kolawole
Hepatitis C virus is one of the emerging infectious diseases that can be transmitted through blood-to-blood contact. This study was carried out to determine the prevalence of anti-HCV antibodies among potential blood donors and pregnant women attending Bowen University Teaching Hospital (BUTH), Ogbomoso, Oyo State. Hospital based study was conducted from December 2014 to September 2015. The study group (N = 279) included potential blood donors and pregnant women. Data on socio-demographic characteristics and potential risk factors were collected using a structured questionnaire...
September 27, 2016: Journal of Immunoassay & Immunochemistry
Elisa María Castells Martínez, Ruben Del Valle, Ernesto Carlos González, Antonio Melchor, Pedro Lucio Pérez, Idania González, Adriana Carr, Kalet León
Human epidermal growth factor is a small peptide consisting of 53 amino acid residues, which stimulates cell proliferation and is associated with several human carcinomas. A simple sandwich-type ultramicroELISA assay (UMELISA), based on the advantages of high affinity reaction between streptavidin and biotin has been developed for the measurement of EGF in human serum samples. Strips coated with a high affinity monoclonal antibody directed against EGF are used as solid phase, to ensure the specificity of the assay...
September 16, 2016: Journal of Immunoassay & Immunochemistry
Jan E Binnie, Delwyn G Cooke, Leonard F Blackwell
An enzyme-linked immunosorbent assay (ELISA) for measurement of pregnanediol-3α-glucuronide (PdG) excretion rates in urine samples diluted to 150 mL/h before analysis is described. The sensitivity of the 9 optimized standard curves was 0.093 ± 0.070 μmol PdG/24 hr, with the multiple combined standard curves having a mean mid-point (EC50) of 6.88 μmol PdG/24 hr. The PdG threshold excretion rate of 7.0 μmol/24 hr, which is used as a marker for the end of fertility, was situated in the most accurate region of the standard curve...
September 16, 2016: Journal of Immunoassay & Immunochemistry
Kheir Eddine Kerboua, Fatma Haiba, Djamila Batouche
Treatment of atypical hemolytic uremic syndrome (aHUS) by the complement C5 inhibitor eculizumab (Soliris®) is highly effective but unfortunately, associated with an economic pressure on the health care systems even in high incomes countries. Despite spacing infusions has been proposed as the unique solution to minimize this economic impact, no reliable laboratory assays are available to tailor such therapy optimization. We aimed to propose and evaluate a complement composite marker for eculizumab efficacy monitoring in aHUS...
September 12, 2016: Journal of Immunoassay & Immunochemistry
David Zeman, Pavlína Kušnierová, Jana Bojková, František Všianský, Olga Zapletalová
IgG kappa and IgG lambda concentrations were quantified in 96 paired CSF and sera using Hevylite™ antibodies in an in-house developed sandwich ELISA method. In 56 of these samples, the results were compared with a qualitative isoelectric focusing/affinity-mediated immunoblotting assay for oligoclonal IgG kappa and IgG lambda. Normal IgG kappa/lambda ratio in the CSF was the same as in serum. In 19/33 patients with intrathecal oligoclonal IgG synthesis, skewed IgG kappa/lambda ratio was observed (increased in 16 and decreased in 3 cases)...
September 9, 2016: Journal of Immunoassay & Immunochemistry
Tsuyoshi Torii, Kentaku Kanemitsu, Akeo Hagiwara
Serum sialic acid (SA) levels are important for diagnosis, follow-up, and mechanistic analysis of malignant diseases. However, little is known about the levels of SA bound to serum IgM. Here, we isolated IgM from sera of healthy individuals and patients with cancer using DEAE chromatography and 8% polyethylene glycol precipitation. In this fraction, which contained partially purified IgM (recovery; 52%; purity: 25%), SA was quantified with fluorescence detection-HPLC (detection limit: 0.08 μM). SA levels in the IgM-enriched fraction was significantly higher in cancer patients (104 ± 27 µM) than in healthy individuals (81 ± 11 µM; p = 0...
September 6, 2016: Journal of Immunoassay & Immunochemistry
Tuba Ozgocer, Sedat Yildiz, Cihat Uçar
Non-invasive measurement of cortisol in saliva is of prime importance as it represents a bioavailable neuroendocrine marker for stress. Therefore, in this study, we developed an enzyme immune assay that was suitable for salivary cortisol measurements. For that purpose, rabbit polyclonal antibody was raised against cortisol-3-CMO:BSA conjugate. The test was based on competition of liquid phase cortisol with conjugated cortisol on the solid phase. Primary antibody was used to bind available sites on the conjugate, which was proportional to numbers of cortisol in liquid phase...
September 6, 2016: Journal of Immunoassay & Immunochemistry
Rafid Mustafa, Elliott Blumenthal
Experiments were conducted to examine the effects of turmeric on spleen cell proliferation. Cell suspensions of spleen cells from young and aged mice were treated with or without conconavalin A (Con-A) as a proliferation stimulant, and with and without turmeric (20 mg/ml) in different concentrations. Spleen cells from young mice that received turmeric showed significant increase in spleen cell proliferation (p<0.05), while spleen cells from aged mice that received turmeric showed no significant increase in T lymphocytes...
August 25, 2016: Journal of Immunoassay & Immunochemistry
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