Development of an immunological method for studying the incorporation of ripening enzymes in cheese curd

1996 ◽  
Vol 63 (1) ◽  
pp. 141-149 ◽  
Author(s):  
Edith Laloy ◽  
Jean-Christophe Vuillemard ◽  
Mouhsine El Abboudi ◽  
Ismael Fliss ◽  
Eric De Grace ◽  
...  
Keyword(s):  
Detection Limit ◽  
Immunosorbent Assay ◽  
Total Protein ◽  
Lactobacillus Casei ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein G ◽  
Immunological Method ◽  
Liposome Suspension ◽  
Cheese Curd

SummaryPolyclonal antibodies raised against partly purified aminopeptidase were specific to cell-free extracts ofLactobacillus caseisubsp.pseudoplantarumUL 137, and did not cross react with any other proteins in Cheddar cheese, at least during the first week of maturation. A sandwich enzyme-linked immunosorbent assay (ELISA) was developed in order to quantify cell-free extracts in the cheese curd, and was also used to determine the efficiency of encapsulation of cell-free extracts in liposomes. This method was very sensitive and exhibited a detection limit of ∼10 μg total protein/g cheese and ∼1 μg total protein/ml liposome suspension. The efficiency of encapsulation of cell-free extracts into liposomes was ∼55–60%. The retention of liposome-encapsulated cell-free extracts was ∼14 times that of non-encapsulated extracts.

scholarly journals Enzyme-Linked Immunosorbent Assay for Simultaneous Detection of Two Fungicides Kresoxim-methyl and Trifloxystrobin in Oranges

2016 ◽  
Vol 34 (No. 5) ◽  
pp. 429-438 ◽  
Author(s):  
Yang Wei ◽  
Zhu Jie ◽  
Liu Ming-Qi ◽  
Dai Xian-Jun
Keyword(s):  
Immunosorbent Assay ◽  
Reaction Rate ◽  
Polyclonal Antibodies ◽  
Simultaneous Detection ◽  
Acid Methyl Ester ◽  
Enzyme Linked Immunosorbent Assay ◽  
Strobilurin Fungicides ◽  
Acid Methyl ◽  
Recovery Study ◽  
The Common

To assemble an indirect competitive enzyme-linked immunosorbent assay (ELISA) for estimating two strobilurin fungicides at the same time, the hapten was synthesised which contained the common active group, (E)-2-(2-bromo-phenyl)-2-(methoxyimino) acetic acid methyl ester (OEBr) in kresoxim-methyl and trifloxystrobin. The immunogen and coating antigen were respectively prepared through conjugating the above-mentioned hapten with BSA and OVA by the mixed anhydride and activated ester methods, and polyclonal antibodies were produced by immunised rabbits. An enzyme-linked immunosorbent assay was developed for simultaneous detection of kresoxim-methyl and trifloxystrobin. In ELISA, the antiserum showed high affinity and sensitivity to kresoxim-methyl and trifloxystrobin, and their IC<sub>50</sub> value and detection limit (expressed as IC<sub>10</sub>) were 14.7 and 0.0044 µg/ml, respectively, for kresoxim-methyl, and 22.9 and 0.014 µg/ml, respectively for trifloxystrobin. The cross-reaction rate was below 0.1% for other strobilurin fungicides. Recovery study of ELISA from spiked samples of homogenised peeled oranges (final concentrations of 100, 10, and 1 µg/ml) resulted in recovery levels in the range of 82–104%.

scholarly journals Easy Enzyme-Linked Immunosorbent Assay for Spectinomycin in Chicken Plasma

1996 ◽  
Vol 79 (2) ◽  
pp. 426-430 ◽  
Author(s):  
Touichi Tanaka ◽  
Hideharu Ikebuchi ◽  
Jun-Ichi Sawada ◽  
Mariko Okada ◽  
Yasumasa Kido
Keyword(s):  
Detection Limit ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Immunosorbent Assay ◽  
Bovine Serum ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Carrier Protein ◽  
Bovine Serum Albumin Conjugate

Abstract An easy, sensitive, competitive indirect enzyme- linked immunosorbent assay (CI-ELISA) for specti nomycin in chicken plasma was developed. Preparation of a spectinomycin-bovine serum albumin conjugate in which the hapten is linked to the carrier protein through the C-4 position is described. Antibodies raised against antigens in rabbits had excellent specificity for spectinomycin, exhibiting a cross-reactivity of 44.0% with dihydrospectinomy-cin and 13.8% with tetrahydrospectinomycin. No cross-reactivity was observed with other antibiotics. The detection limit of the CI-ELISA was 2 ng/mL (equivalent into 40 ng/mL undiluted chicken plasma) spectinomycin. Known amounts (0.1-100 μg/mL) of spectinomycin were added to chicken plasma and then analyzed. Average recoveries were 97-110%. This procedure may be used without prior extraction of samples.

Quantification of serum amyloid P by enzyme-linked immunosorbent assay

1991 ◽  
Vol 37 (10) ◽  
pp. 1742-1745 ◽  
Author(s):  
R Saïle ◽  
A Kandoussi ◽  
M Deveaux ◽  
J Descamps ◽  
E Hachulla ◽  
...  
Keyword(s):  
Human Plasma ◽  
Immunosorbent Assay ◽  
Polyclonal Antibodies ◽  
Serum Amyloid ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Amyloid P ◽  
Assay Procedure ◽  
Normal Individuals ◽  
Sandwich Type ◽  
Amyloid P

Abstract This enzyme-linked immunosorbent assay procedure for quantifying serum amyloid P (SAP) in human plasma makes use of affinity-purified polyclonal antibodies to SAP in a "sandwich"-type format. The procedure is sensitive, reproducible, simple, and easily automatable. Results correlate well with those by a rocket immunoelectrophoresis method performed with the same antibodies. Sera from apparently normal individuals had a mean SAP content of 44.17 mg/L and increased with age.

Development of a Highly Sensitive Enzyme-Linked Immunosorbent Assay Based on Polyclonal Antibodies for the Detection of Polychlorinated Dibenzo-p-dioxins

1998 ◽  
Vol 70 (6) ◽  
pp. 1092-1099 ◽  
Author(s):  
Yukio Sugawara ◽  
Shirley J. Gee ◽  
James R. Sanborn ◽  
S. Douglass Gilman ◽  
Bruce D. Hammock
Keyword(s):  
Immunosorbent Assay ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Highly Sensitive

Development of Enzyme Immunoassay for Captan and Its Degradation Product Tetrahydrophthalimide in Foods

1993 ◽  
Vol 76 (2) ◽  
pp. 381-386 ◽  
Author(s):  
W Harvey Newsome ◽  
Jupiter M Yeung ◽  
Peter G Collins
Keyword(s):  
Detection Limit ◽  
Human Serum Albumin ◽  
Degradation Product ◽  
Protein Concentration ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Test Compound ◽  
Coefficients Of Variation ◽  
Spacer Arm ◽  
Related Compounds

Abstract A simple, sensitive, and precise enzyme-linked immunosorbent assay (ELISA) is described for the quantitation of captan as its degradation product tetrahydrophthalimide (THPI) in foods using polyclonal antibodies. Three hapten analogues of THPI with different alky I spacer arm lengths were synthesized. Immunogens and coating proteins were prepared by coupling these haptens to human serum albumin and ovalbumin, respectively. A 5-carbon spacer arm appeared to be optimum for the production of antibodies. Heterologous coating proteins did not improve the sensitivity, but reduction of homologous coating protein concentration did improve the sensitivity, resulting in a concentration of test compound required to inhibit binding by 50% of 15.5 ng/mL The antiserum is specific for captan, captafol, and THPI, but not other structurally related compounds. The minimum detection limit was 1 ng/mL; the linearity was 1-200 ng/mL. The overall recoveries of captan and THPI from 11 commodities spiked at 4 levels were 92 and 100%, respectively. The intra-assay and interassay coefficients of variation were 9.1 and 16.8% for apple blanks and 5.9 and 4.2% for apple spiked with 3 ppm THPI, respectively. The ELISA described is suitable for measuring captan and THPI at levels comparable to those typically found in fruit.

Production and Characterization of Antibodies against Fumonisin B1

1996 ◽  
Vol 59 (9) ◽  
pp. 992-997 ◽  
Author(s):  
FENG-YIR YU ◽  
FUN S. CHU
Keyword(s):  
Detection Limit ◽  
Polyclonal Antibodies ◽  
Fumonisin B1 ◽  
Correlation Coefficients ◽  
Keyhole Limpet Hemocyanin ◽  
Hplc Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Indirect Competitive Elisa ◽  
Antibody Titers

Polyclonal antibodies against fumonisin B1 (FmB1) were produced in rabbits after immunizing the animals with either FmBl-keyhole limpet hemocyanin (KLH) or FmB1 bovine serum albumin (BSA). A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) and an indirect competitive ELISA (idc-ELISA) were used for the characterization of the antibodies and for analysis of the toxin in corn samples. The antibody titers in the serum of rabbits immunized with FmBl-KLH were considerably higher than in those immunized with FmBl-BSA. The antibodies from the rabbits immunized with FmBl-KLH were further characterized. The concentrations causing 50% inhibition of binding of FmB1-horseradish peroxidase (HRP) to the antibodies by FmB1, FmB2 and FmB3 in the ELISA were found to be 0.45, 0.72, and 25 ng/ml, respectively. The detection limit of FmBl, based on 95% confidence at 5% of inhibition of binding of FmBl-HRP conjugate, in buffer of the dc-ELISA was found to be 0.05 ng/ml. In the presence of a matrix such as corn, the detection limit was less than 50 ppb. The overall analytical recoveries of FmBl (50 to 1,000 ng/g) added to the ground corn and then extracted with CH3CN/H2O (1/1, vol/vol) with cleanup and without cleanup in the dc ELISA were found to be 70.5 and 85.9%, respectively. A good correlation was found between the FmBl levels in 2 starch and 10 naturally contaminated corn samples analyzed by the dc-ELISA and the high-pressure liquid chromatography (HPLC) method. The correlation coefficients between ELISA and HPLC were found to be 0.955 (y [ELISA] = 1.3 1x [HPLC] + 77 ppb; P &lt; 0.001) and 0.811 (y = 1.13x + 34 ppb; P &lt; 0.01) for the sample without and with cleanup treatment, respectively.

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