Prostatic inhibin-like peptide quantified in urine of prostatic cancer patients by enzyme-linked immunosorbent assay.

1989 ◽  
Vol 35 (7) ◽  
pp. 1376-1379 ◽  
Author(s):  
T R Teni ◽  
A H Bandivdekar ◽  
A R Sheth ◽  
N A Sheth
Keyword(s):  
Cancer Patients ◽  
Immunosorbent Assay ◽  
Prostatic Cancer ◽  
Solid Phase ◽  
Microtiter Plate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Analytical Recovery ◽  
The Status ◽  
Polystyrene Microtiter Plate ◽  
The Mean

Abstract This is a highly specific enzyme-linked immunosorbent assay (ELISA) for measuring prostatic inhibin-like peptide (PIP) in urine, in which we use penicillinase (EC 3.5.2.6) conjugated with PIP and, as solid phase, a polystyrene microtiter plate. We used this ELISA to measure PIP in 24-h urine specimens from men with prostatic cancer (PCa) and from age-matched controls. For prostatic cancer patients the mean +/- SEM urinary PIP of 36.1 +/- 5 micrograms/24 h was significantly (P less than 0.001) lower than the mean of 127.1 +/- 9 micrograms/24 h for the age-matched controls. PIP values for 30 samples measured by both ELISA and RIA correlated well (r = 0.985). We could detect as little as 1.56 ng of PIP in a sample. Analytical recovery of added PIP ranged from 91% to 104%. Mean CVs were 8.9% within-assay and 12.7% between-assay. We believe that this ELISA will be useful in assessing the status of PIP in men with normal and diseased prostates and in examining the function of the hypothalamus-pituitary-prostate axis.

Indirect Enzyme-Linked Immunosorbent Assay for Saxitoxin in Shellfish

1985 ◽  
Vol 68 (1) ◽  
pp. 13-16 ◽  
Author(s):  
Fun S Chu ◽  
Titan S L Fan
Keyword(s):  
Detection Limit ◽  
Immunosorbent Assay ◽  
Indirect Elisa ◽  
Solid Phase ◽  
Microtiter Plate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chromogenic Substrate ◽  
Rabbit Igg ◽  
Lower Detection ◽  
Peroxidase Conjugate

Abstract An indirect enzyme-linked immunosorbent assay (ELISA) was developed for the detection of saxitoxin (STX). Antibodies against STX were demonstrated in rabbits 5 weeks after immunizing with STX-bovine serum albumin (STX-HCHO-BSA). In the ELISA, STX-HCHO-BSA or polylysine-STX was coated onto the microtiter plate, followed by incubation with standard toxin and anti-STX antibody. The amount of antibody bound to the solid phase was determined by incubation with goat anti-rabbit IgG peroxidase conjugate and a reaction with chromogenic substrate. Competitive indirect ELISA revealed that the antiserum did not cross-react with either carbamoyl-neo-STX-suIfate or tetrodotoxin. The antibodies for STX cross-reacted with decarbamoyl- STX and neo-STX about 56% and 16% as much as they did with STX, respectively. The lower detection limits for STX, decarbamoyl-STX, and neo-STX in this sytem were about 25, 45, and 156 pg per assay, respectively. When STX added to clams or mussels was assayed, the detection limit for STX was about 50-100 ppb, and recoveries were in the range of 86.8-107%.

Participation of peptide moieties in adhesive behavior of antigenic mannans of Candida albicans to the plastic microtiter plate in enzyme-linked immunosorbent assay.

1987 ◽  
Vol 33 (10) ◽  
pp. 1925-1928 ◽  
Author(s):  
M Tojo ◽  
N Shibata ◽  
T Mikami ◽  
M Suzuki ◽  
S Suzuki
Keyword(s):  
Candida Albicans ◽  
Immunosorbent Assay ◽  
Alkaline Solution ◽  
Yeast Species ◽  
Microtiter Plate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Aqueous Alkaline Solution ◽  
Pathogenic Yeast ◽  
Color Development ◽  
Polystyrene Microtiter Plate

Abstract We report our studies of the mechanism of the adhesion of mannans of Candida albicans NIH A-207 and C. albicans NIH B-792 strains to the surface of a polystyrene microtiter plate being utilized for enzyme-linked immunosorbent assay (EIA) of antigens of this pathogenic yeast species. This binding was manifested predominantly by the peptide moieties of the mannans forming hydrophobic bonds with the plastic molecule. Eliminating the peptide of each mannan by treating it with a hot aqueous alkaline solution of NaBH4 also eliminated color development in the EIA.

Enzyme-Linked Immunosorbent Assay of Aflatoxin Bi in Naturally Contaminated Corn and Cottonseed

1986 ◽  
Vol 69 (5) ◽  
pp. 904-907 ◽  
Author(s):  
Bhanu P Ram ◽  
L Patrick Hart ◽  
Odette L Shotwell ◽  
James J Pestka
Keyword(s):  
Correlation Coefficient ◽  
Immunosorbent Assay ◽  
Solid Phase ◽  
Microtiter Plate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Extraction Solvent ◽  
Sample Extract ◽  
Coefficients Of Variation ◽  
Layer Chromatography ◽  
Duplicate Sample

Abstract Naturally contaminated corn and cottonseed samples were screened for aflatoxin B1 (AFB1) by a direct competitive enzyme-linked immunosorbent assay (ELISA). Samples were blended 5 min in an extraction solvent of methanol-water-dimethylformamide (70 + 29 + 1) and filtered. Filtrates were assayed by direct competition between AFBi in the corn and cottonseed samples and AFB1-peroxidase conjugate for binding to specific antibody adsorbed to a solid phase microtiter plate. Standard curves prepared using the extract of AFB1-free corn and cottonseed samples, and extraction solvent only, showed negligible interference by the sample extract in the performance of ELISA. The AFBi content in corn and dehulled cottonseed samples as determined by ELISA ranged from 7 to 422 μg/kg and 7 to 3258 μg/kg, respectively. When ELISA estimates of AFB1 in corn were compared with values obtained by thin layer chromatography (CB method), the correlation coefficient (n = 10) was 0.95. Average interassay and subsample coefficients of variation for ELISA in corn were 21.4 and 22.0%, respectively. When ELISA estimates of AFB1 in cottonseed were compared with values obtained by liquid chromatography (Pons method), the correlation coefficient (n = 15) was 0.96. Using this ELISA, 36 duplicate sample extracts can be screened for AFB1 in less than 2 h.

Competitive time-resolved immunofluorometric assay for quantifying carbonic anhydrase VI in saliva

1993 ◽  
Vol 39 (10) ◽  
pp. 2154-2157 ◽  
Author(s):  
S Parkkila ◽  
A K Parkkila ◽  
T Vierjoki ◽  
T Ståhlberg ◽  
H Rajaniemi
Keyword(s):  
Carbonic Anhydrase ◽  
Solid Phase ◽  
Microtiter Plate ◽  
Time Resolved ◽  
Solid Phase Immunoassay ◽  
Resolution Principle ◽  
Rabbit Igg ◽  
Analytical Recovery ◽  
The Mean ◽  
Carbonic Anhydrase Vi

Abstract A competitive time-resolved immunofluorometric assay sensitive and robust enough for quantifying human salivary carbonic anhydrase isoenzyme VI (HCA VI) was developed. The solid-phase immunoassay is based on competition between Eu(3+)-labeled HCA VI and salivary HCA VI for polyclonal rabbit anti-HCA VI antibodies that are attached to microtiter plate wells precoated with sheep anti-rabbit IgG. The subsequent immunoassay including the separation of free and bound HCA VI requires only one incubation step, after which the Eu3+ of the bound labeled antigen is released into an enhancement solution. The highly fluorescent Eu chelates formed in this solution are then quantified by time-resolved fluorometry (Delfia). The time-resolution principle effectively obviates possible interferences from complex biological material such as saliva. The assay detection limit was 1.5 micrograms/L. Intra- and interassay imprecisions (CVs) were 5.1% and 5.3%, respectively. The mean analytical recovery was 93%. The mean +/- SD concentration of HCA VI in paraffin-stimulated saliva was 6.8 +/- 4.3 mg/L (n = 30) and the secretion rate was 10.2 +/- 7.9 micrograms/min. The method was useful for further investigations of the role of HCA VI in difficult matrices, e.g., saliva.

New enzyme-linked immunosorbent assay for glycocalicin in plasma

1991 ◽  
Vol 37 (2) ◽  
pp. 169-172 ◽  
Author(s):  
ShinjI Kunishima ◽  
Klyotaka Hayashi ◽  
Sentaro Kobayashi ◽  
Tomokl Naoe ◽  
Ryuzo Ohno
Keyword(s):  
Immunosorbent Assay ◽  
Binding Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Glycoprotein Ib ◽  
Proteolytic Fragment ◽  
Competitive Binding Assay ◽  
Analytical Recovery ◽  
Sandwich Type ◽  
Assay Time ◽  
The Mean

Abstract A new sandwich-type enzyme-linked immunosorbent assay for quantifying glycocalicin, a proteolytic fragment of platelet membrane glycoprotein Ib, is described. The assay is based on the use of two monoclonal antibodies raised against glycoprotein Ib and involves the avidin-biotin technique. The detection limit is 7 micrograms/L and the range of glycocalicin determined in plasma is 0.01 to 1 mg/L. Assay time is 2 h. The intra-assay CV ranged from 3.6% to 5.2%, the interassay CV from 5.4% to 8.0%. Analytical recovery of purified glycocalicin added to a plasma pool averaged 96%. In 36 healthy subjects, the mean glycocalicin concentration in plasma was 0.36 (SD 0.07) mg/L (2.7 nmol/L). We conclude that this assay is suitable for measuring glycocalicin in plasma and is also more sensitive and precise than the previously published immunoassays based on competitive binding assay.

SENSITIVE AND SPECIFIC ENZYME-LINKED IMMUNOSORBENT ASSAY FOR UROKINASE-TYPE PLASMINOGEN ACTIVATOR IN HUMAN PLASMA

1987 ◽  
Author(s):  
J Grøndahl-HANSEN ◽  
N Agerlin ◽  
L S Nielsen ◽  
K Danø
Keyword(s):  
Plasminogen Activator ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mean Values ◽  
Amino Terminal ◽  
Type Plasminogen Activator ◽  
Healthy Donors ◽  
Urokinase Type Plasminogen Activator ◽  
The Mean

An enzyme-linked immunosorbent assay (ELISA) was developed for the measurement of human urokinase-type plasminogen activator (u-PA) in plasma and serum. Microtiter plates were coated with a monoclonal antibody and incubated with standard or sample. Bound u-PA was quantitated with polyclonal antibodies conjugated with biotin, followed by avidin-peroxidase. The assay was 10-fold as sensitive as other previously reported ELISAs, the detection limit being approximately 1 pg of u-PA in a volume of 100 μl with a linear dose-response up to 15 pg of u-PA. The assay detected active u-PA and its inactive proenzyme form equally well and the recovery of both forms was higher than 90% in plasma. A variety of structurally related proteins, including t-PA, were tested, but no reaction with proteins other than u-PA and its amino-terminal degradation product were observed. The intra-assay and inter-assay coefficients of variation for determination of u-PA in plasma were 7.6% and 8.4%, respectively. The assay was equally applicable to serum. The values obtained with plasma and serum were similar, and the results were not affected by small variations in the preparation of the samples. The ELISA was used to measure the concentration of u-PA in plasma from 34 healthy donors. The mean values for u-PA in plasma from healthy donors was 1.1 ng/ml ± 0.3 ng/ml (SD) (range 0.6 - 1.5 ng/ml). No significant differences were found between men and women and no correlation between u-PA concentration and age could be demonstrated.The mean u-PA concentration in plasma from healthy donors obtained in this study is substantially lower than that reported by others. This might be due to different methods of determination of the protein content of the standard preparations or to differences in the specificity of the assays.

A New Technique in Quantitative Immunohematology: Solid-Phase Kinetic Enzyme-Linked Immunosorbent Assay

Vox Sanguinis ◽  
1983 ◽  
Vol 45 (6) ◽  
pp. 440-448 ◽  
Author(s):  
S. Spitalnik ◽  
J. Cowles ◽  
M.T. Cox ◽  
D. Baker ◽  
J. Holt ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Solid Phase ◽  
New Technique ◽  
Enzyme Linked Immunosorbent Assay ◽  
A New Technique

scholarly journals Antibody level of New Zealand children immunized with the triple vaccine DTP (diphtheria-tetanus-pertussis)

1988 ◽  
Vol 101 (2) ◽  
pp. 405-410 ◽  
Author(s):  
R. C. H Lau
Keyword(s):  
New Zealand ◽  
Immunosorbent Assay ◽  
Antibody Level ◽  
Herd Immunity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibody ◽  
Immunization Strategy ◽  
The Mean ◽  
Antibody Levels

SUMMARYEnzyme-linked immunosorbent assay (ELISA) tests were used to measure IgG antibody levels in 2638 New Zealand children who had been immunized with the triple vaccine DTP. The percentage of children immune to diphtheria decreased with age. The percentage of children immune to tetanus varied from 67.1 to 55.0%. The percentage of children with measurable antibody to pertussis increased with age. The mean percentages of children with measurable antibody or immunity to one or more DTP components were 34.2% (with 3 components), 34.4% (2 components), and 78.1% (1 component). It appears the immunization strategy for diphtheria and tetanus is satisfactory for herd immunity in New Zealand children. However, the current pertussis strategy may not be providing adequate immunity to 5-year-olds in this country.

scholarly journals Mycotoxins as a risk in the grain food

2009 ◽  
pp. 79-86 ◽  
Author(s):  
Jovana Matic ◽  
Jasna Mastilovic ◽  
Ivana Cabarkapa ◽  
Anamarija Mandic
Keyword(s):  
European Union ◽  
Secondary Metabolites ◽  
Ochratoxin A ◽  
Immunosorbent Assay ◽  
Toxic Effects ◽  
Enzyme Linked Immunosorbent Assay ◽  
The European Union ◽  
The Mean ◽  
Grain Foods

Mycotoxins are toxic secondary metabolites of fungi that contaminate a large variety of foods and have toxic effects on humans. The best protection against mycotoxins is to monitor their presence in food. This paper shows the screening results of mycotoxins present in 76 samples of different groups of grain foods. Samples of grain food were analyzed for contamination with aflatoxins, ochratoxin A, zearalenone, fumonisins and deoxynivalenol. Analysis were conducted using competitive enzyme-linked immunosorbent assay (ELISA). None of the samples was contaminated with aflatoxins. The most predominant mycotoxin was ochratoxin A with the mean level of 4.84 ? 4.49 ppb in 19.7% of the examined samples. Zearalenone, fumonisins, and deoxynivalenol were found in 9.21, 14.5 and 3.9% of the samples, respectively. Mycotoxin content in the investigated samples was compared with the regulations of Serbia and those of the European Union.

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