Indirect Competitive Enzyme Immunoassay for Tetrodotoxin Using a Biotin-Avidin System

1992 ◽  
Vol 75 (5) ◽  
pp. 883-886 ◽  
Author(s):  
Kendo Matsumura ◽  
Sadako Fukiya
Keyword(s):  
Bovine Serum Albumin ◽  
Enzyme Immunoassay ◽  
Immunoglobulin G ◽  
Bovine Serum ◽  
Keyhole Limpet Hemocyanin ◽  
Microtiter Plate ◽  
Mouse Bioassay ◽  
Detection Level ◽  
Standard Curve ◽  
Peroxidase Conjugate

Abstract An indirect competitive enzyme immunoassay was developed to determine tetrodotoxin (TTX). Antiserum against TTX was demonstrated in rabbits immunized with TTX-keyhole limpet hemocyanin conjugate. In this assay, TTX-bovine serum albumin was coated on the microtiter plate, which was incubated with standard TTX and biotinylated immunoglobulin G from antiserum. The amount of antibody bound on the surface of wells was determined by the reaction of avidin-peroxidase conjugate with its substrate. The minimal detection level of TTX ranged from 5 to 25 pg/assay. The standard curve was linear between 0.25 and 50 ng/assay. A good correlation (r= 0.974) for TTX was also obtained between this indirect competitive enzyme immunoassay and the mouse bioassay.

Sensitive Enzyme Immunoassay for Anti-β-Lactoglobulin IgG in Serum

1998 ◽  
Vol 35 (5) ◽  
pp. 649-655
Author(s):  
Kouichi Hirota ◽  
Seiichi Hashida ◽  
Eiji Ishikawa ◽  
Masayuki Totani
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Enzyme Immunoassay ◽  
Peroxidase Activity ◽  
Immunoglobulin G ◽  
Bovine Serum ◽  
Healthy Subjects ◽  
Enzyme Linked Immunosorbent Assay ◽  
Β Lactoglobulin ◽  
Peroxidase Conjugate

A sensitive enzyme immunoassay for anti-β-lactoglobulin immunoglobulin G (IgG) in serum is described. Serum containing anti-β-lactoglobulin IgG was reacted simultaneously with 2,4-dinitrophenyl-bovine serum albumin-β-lactoglobulin conjugate and β-lactoglobulin-peroxidase conjugate. The complex formed from the three components was trapped onto polystyrene balls coated with anti-2,4-dinitrophenyl group IgG, eluted with ε-2,4-dinitrophenyl-L-lysine and transferred to polystyrene balls coated with anti-human IgG-γ-chain IgG. Bound peroxidase activity was determined by fluorometry. This enzyme immunoassay was 100- to 1000-fold more sensitive and more reliable than the enzyme-linked immunosorbent assay (ELISA). Anti-β-lactoglobulin IgG was detected in 91% of healthy subjects using this method.

scholarly journals KONJUGASI ANHIDRAT KlORAMFENIKOL-PROTEIN (CAP-BSA DAN CAP-KLH) UNTUK PRODUKSI IgG DARI SERUM KELINCI

2011 ◽  
Vol 1 (2) ◽  
pp. 23-31
Author(s):  
Musyirna Rahmah Nst ◽  
Tri Budhi Murdiati
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Immunosorbent Assay ◽  
Bovine Serum ◽  
Keyhole Limpet Hemocyanin ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein Bovine Serum Albumin

 Dalam pengembangan metoda analisis residu antibiotik pada produk pangan hewani secara immunokimia telah dilakukan sintesis imunogen kloramfenikol (CAP) dengan protein Bovine Serum Albumin (BSA) dan Keyhole Limpet Hemocyanin (KLH) dengan metoda konjugasi mixed anhidrad. Hasil sintesis digunakan untuk produksi antibodi.Antigen dan antibodi yang dihasilkan dijadikan prangkat analisis residu antibiotik dengan Competitive Enzyme Linked Immunosorbent Assay (ELISA kompetitif). Hasil penelitian menunjukkan bahwa metoda konjugasi mixed anhidrad pada preparasi imunogen dengan jumlah CAP yang terkonjugasi ke grup amino bebas dari molekul protein BSA (CAP-BSA) adalah 18 unit dan ke protein KLH (CAP-KLH) adalah 782 unit. Antibodi poliklonal CAP-BSA dan CAP-KLH yang diproduksi dengan rata-rata kandungan IgG CAP-BSA 2,07 mg/ml dan IgG CAP-KLH 2,21 mg/ml.

Sensitive Enzyme Immunoassay of Cephalexin Residues in Milk, Hen Tissues, and Eggs

1988 ◽  
Vol 71 (5) ◽  
pp. 915-920
Author(s):  
Tsunehiro Kitagawa ◽  
Yukio Gotoh ◽  
Kazuyo Uchihara ◽  
Youko Kohri ◽  
Tihoko Kinoue ◽  
...  
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Enzyme Immunoassay ◽  
Bovine Serum ◽  
Limit Of Detection ◽  
Rabbit Antiserum ◽  
Egg Yolk ◽  
Amino Group ◽  
Thiol Groups ◽  
Cross Linker

Abstract A sensitive enzyme immunoassay for cephalexin (CEX) was developed using the rabbit antiserum to CEX, Β-D-galactosidase-labeled CEX, and a double-antibody separation method. The immunogen of CEX was prepared by coupling the amino group of CEX to thiol groups introduced into bovine serum albumin by the use of N-(mmaleimidobenzoyloxy) succinimide as a cross-linker. Highly titered antiserum to CEX was produced in rabbits immunized with the immunogen. Enzyme labeling of CEX with Β-D-galactosidase was done by using 7V-(gamma-maleimidobutyryloxy)succinimide as the crosslinker. The limit of detection was 30 ng CEX/mL sample solution. Application of the method to CEX drug residues detected 30 ng/mL in milk, 60 ng/g in egg yolk, and 400 ng/g in hen tissue.

scholarly journals ENZYME IMMUNOASSAY KIT FOR THE DETERMINATION OF DEOXYNIVALENOL

2018 ◽  
Vol 54 (3) ◽  
pp. 319-328 ◽  
Author(s):  
I. I. Vashkevich ◽  
O. S. Kuprienko ◽  
I. V. Gorbachova ◽  
A. A. Yastrebova ◽  
T. V. Terentieva ◽  
...  
Keyword(s):  
Bovine Serum Albumin ◽  
Horseradish Peroxidase ◽  
Quantitative Determination ◽  
Enzyme Immunoassay ◽  
Bovine Serum ◽  
Polyclonal Antibodies ◽  
Agricultural Products ◽  
Accuracy And Precision ◽  
Modern Level

A reagent kit EIA-DEOXYNIVALENOL for the determination of mycotoxin deoxynivalenol (DON) in feeds and foods by a direct competitive enzyme immunoassay using microtitration plate has been developed and tested. The basic components of the kit are polyclonal antibodies to DON, obtained as a result of immunization of rabbits with a conjugate of DON with bovine serum albumin and a conjugate of horseradish peroxidase with DON. The evaluated parameters of the kit and metrological characteristics of the technique of measurements correspond to the modern level of immunoassay development and provide the determination of DON content of agricultural products in a range of 0.2 to 6.0 mg/kg with proper accuracy and precision. The limit of quantitative determination of DON in grain and cereal foods does not exceed 0.2 mg/kg.

Microencapsulated antibodies in radioimmunoassay. III. Determination of cortisol.

1980 ◽  
Vol 26 (5) ◽  
pp. 633-634 ◽  
Author(s):  
R W Bordens ◽  
E P Halpern
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Plasma Cortisol ◽  
Total Plasma ◽  
Competitive Reaction ◽  
Large Molecules ◽  
Standard Curve ◽  
Analytical Recovery

Abstract We describe a method for directly measuring total plasma cortisol by use of a microencapsulated antibody. The antibody microcapsules were incubated with plasma and 125I-labeled cortisol in a competitive reaction at 37 degrees C for 15 min, then separated by centrifugation; the radioactivity of the pellet was counted for 1 min. Analytical recovery of cortisol from three plasma pools was 97.6% (SD 8.5%) and was unaffected by triglyceridemia, hemolysis, or icterus. The standard curve was linear to 500 microgram/L and the sensitivity was 10 microgram/l. Recovery of cortisol added as the hemisuccinate was 102% (SD 10.6%), whereas that of the conjugate cortisol hemisuccinate/bovine serum albumin was 4.3% (SD 3.5%). This confirmed that the microcapsule excludes large molecules. The within-day CV for two control pools was 9.8 and 12.8%, the day-to-day variation 13.4 and 13.8%.

The Solubility of Insect Juvenile Hormone in Aqueous Solutions and Its Adsorption by Glassware and Plastics

1977 ◽  
Vol 32 (3-4) ◽  
pp. 158-160 ◽  
Author(s):  
Ch. Giese ◽  
K. D. Spindler ◽  
H. Emmerich
Keyword(s):  
Ionic Strength ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Aqueous Solutions ◽  
Juvenile Hormone ◽  
Immunoglobulin G ◽  
Bovine Serum ◽  
Plastic Materials ◽  
Buffer Composition ◽  
Ph Buffer

Abstract The C18 juvenile hormone of insects can be dissolved in aqueous solutions up to a concentration of 2.5 - 3.0 × 10-5 M; changes in pH, buffer composition and ionic strength hardly affect this solu­bility. The hormone is salted out gradually by increasing ammonium sulphate concentrations. The juvenile hormone is bound to proteins such as bovine serum albumin or goat immunoglobulin G and can be kept in solution by these proteins up to 10-3 M. The hormone is strongly absorbed by many commonly used plastic materials but only to a lesser extent by glass and teflon.

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