Human corticotropin (ACTH) radioimmunoassay with synthetic 1--24 ACTH.

1979 ◽  
Vol 25 (7) ◽  
pp. 1267-1273 ◽  
Author(s):  
P C Kao ◽  
N S Jiang ◽  
P C Carpenter
Keyword(s):  
Polyethylene Glycol ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Confidence Limit ◽  
Normal Subjects ◽  
Cross Reactivity ◽  
Normal Range ◽  
Beta Endorphin

Abstract A corticotropin antiserum was obtained from rabbits immunized with synthetic 1--24 corticotropin conjugated with bovine serum albumin. The antiserum did not cross react with synthetic alpha-melanotropin or with synthetic beta-endorphin and had a cross reactivity of 0.23% with human beta-lipotropin. We developed a radioimmunoassay with the antiserum obtained, in which we used polyethylene glycol in conjunction with a second precipitating antibody for fast (15-min) separation of antibody-bound and free corticotropin. The assay had a sensitivity of 16 ng/L and was validated on patients with various pituitary and adrenal diseases. From 103 normal subjects, the median value for corticotropin in specimens collected during the morning was 34 ng/L of plasma; the upper 95% confidence limit of the normal range was 98 ng/L.

Production and Characterization of Antibody Against 3′-OH-T-2 Toxin

1986 ◽  
Vol 49 (4) ◽  
pp. 267-271 ◽  
Author(s):  
RU-DONG WEI ◽  
WILLIAM BISCHOFF ◽  
FUN SUN CHU
Keyword(s):  
Detection Limit ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Cross Reactivity ◽  
Toxin A

Antibody raised against T-2 toxin cross-reacted poorly with 3′-OH-T-2 toxin. A new immunogen was prepared by conjugation of hemisuccinate (HS) of 3′-OH-T-2 toxin to bovine serum albumin (BSA). Antibodies against 3′-OH-T-2 toxin were demonstrated by a radioimmunoassay 10 wk after immunization of rabbits with this new immunogen using tritiated 3′-OH-T-2 toxin as the testing ligand. Highest titers (1:6,000) were obtained 17 wk after immunization and two booster injections. The antibodies had good cross-reactivity with T-2 toxin, acetyl-T-2 toxin and 3′-OH-acetyl-T-2 toxin. The relative cross-reactivity of this antibody with 3′-OH-T-2, acetyl-T-2, T-2, 3′-OHacetyl-T-2, 3′-OH-T-2-HS, T-2 isomer, HT-2 and 3′-OH-HT-2 was 1, 3, 4, 5, 15, 30, 45 and 175, respectively. No crossreaction was found when 3′-OH-T-2 triol, T-2-triol, T-2-tetraol, DAS and DON at a concentration of 1 μg per assay was tested. The detection limit for 3′-OH-T-2 toxin by the RIA was about 0.1 ng per assay.

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.

scholarly journals Heterologous screening of hybridomas for the development of broad-specific monoclonal antibodies against deoxynivalenol and its analogues

2014 ◽  
Vol 7 (3) ◽  
pp. 257-265 ◽  
Author(s):  
Y. Guo ◽  
M. Sanders ◽  
A. Galvita ◽  
A. Heyerick ◽  
D. Deforce ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Bovine Serum Albumin ◽  
Horseradish Peroxidase ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Simultaneous Detection ◽  
Cross Reactivity ◽  
Efficient Approach ◽  
Selection For ◽  
Similar Affinity

Hapten heterology was introduced into the steps of hybridoma selection for the development of monoclonal antibodies (MAbs) against deoxynivalenol (DON). Firstly, a novel heterologous DON hapten was synthesised and covalently coupled to proteins (i.e. bovine serum albumin (BSA), ovalbumin and horseradish peroxidase) using the linkage of cyanuric chloride (CC). After immunisation, antisera from different DON immunogens were checked for the presence of useful antibodies. Next, both homologous and heterologous enzyme-linked immunosorbent assays were conducted to screen for hybridomas. It was found that heterologous screening could significantly reduce the proportion of false positives and appeared to be an efficient approach for selecting hybridomas of interest. This strategy resulted in two kinds of broad-selective MAbs against DON and its analogues. They were quite distinct from other reported DON-antibodies in their cross-reactivity profiles. A unique MAb 13H1 derived from DON-CC-BSA immunogen could recognise DON and its analogues in the order of HT-2 toxin ≯ 15-acetyl-DON ≯ DON ≯ nivalenol, with IC50 ranging from 1.14 to 7.69 μg/ml. Another preferable MAb 10H10 generated from DON-BSA immunogen manifested relatively similar affinity to DON, 3-acetyl-DON and 15-acetyl-DON, with IC50 values of 22, 15 and 34 ng/ml, respectively. This is the first broad-specific MAb against DON and its two acetylated forms and thus it can be used for simultaneous detection of the three mycotoxins.

Radioimmunoassay for 13, 14-dihydro-15-ketoprostaglandin F2α and its application in normo- and anovulatory women

1982 ◽  
Vol 100 (1) ◽  
pp. 98-104 ◽  
Author(s):  
Werner Schlegel ◽  
Jaime Urdinola ◽  
Hermann P. G. Schneider
Keyword(s):  
Menstrual Cycle ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Rapid Method ◽  
Plasma Levels ◽  
Bovine Serum ◽  
Limit Of Detection ◽  
Cross Reactivity ◽  
Specific Antibodies ◽  
Standard Range

Abstract. Highly specific antibodies to 13, 14-dihydro-15-ketoprostaglandin F2α (PGFM) were raised in rabbits. The animals were immunized with PGFM-bovine serum albumin (BSA)-conjugates. Prior to the incubation procedure PGFM was extracted by a rapid method with dichloromethane followed by column chromatography. The antisera dilution was 1:10000 and the cross-reactivity towards prostaglandin A2, E2, F2α, 13, 14-dihydro-15-ketoprostaglandin E2 and the 15-ketoprostaglandin E2 and F2α was < 1%. The limit of detection was 1.9 ± 0.6 pg/ml plasma over the standard range 1.9–250 pg. The intra- and inter-assay variations were 3.9 and 15%, respectively. PGFM was measured throughout the menstrual cycle in female volunteers. In normal ovulatory women (n = 3) plasma levels of PGFM varied between 65.6 to 107.1 pg/ml. No significant variations of plasma PGFM were seen during the cycle. In anovulatory women (n = 4) no difference of PGFM was found during the cycle. PGFM levels in hyperprolactinaemic but ovulating women tend to be higher than in anovulatory, and normoprolactinaemic subjects. These data strongly indicate that PGFM is not correlated with other hormonal parameters tested here in the normal and anovulatory cycles.

Lack of specificity of current anti-digoxin antibodies, and preparation of a new, specific polyclonal antibody that recognizes the carbohydrate moiety of digoxin.

1985 ◽  
Vol 31 (10) ◽  
pp. 1625-1631 ◽  
Author(s):  
B Thong ◽  
S J Soldin ◽  
C A Lingwood
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Polyclonal Antibody ◽  
Bovine Serum ◽  
Lactone Ring ◽  
Cross Reactivity ◽  
Carbohydrate Moiety ◽  
Reductive Ozonolysis ◽  
Bovine Serum Albumin Conjugate ◽  
Specific Polyclonal Antibody

Abstract Current immunoassays for digoxin do not distinguish digoxin from its glycosidic metabolites. We have synthesized a novel digoxin/bovine serum albumin conjugate via reductive ozonolysis of the lactone ring such that the carbohydrate moiety of digoxin remains intact. Antibodies raised against this conjugate show minimal cross reactivity to digoxigenin, bisdigitoxide, monodigitoxide, digoxigenin, and digitoxin. With this antibody, digoxin can be measured in the presence of these metabolites.

Production and Characterization of Antibodies Against Neosaxitoxin

1992 ◽  
Vol 75 (2) ◽  
pp. 341-345 ◽  
Author(s):  
Fun S Chu ◽  
Xuan Huang ◽  
Sherwood Hall
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Indirect Elisa ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
High Antibody ◽  
Antibody Titers ◽  
Antibody Elisa

Abstract Antibody against neosaxitoxin (neo-STX) was obtained from rabbits after immunization with neo- STX conjugated to either keyhole limpet hemocyanln (KLH) or bovine serum albumin (BSA). An indirect enzyme-linked Immunosorbent assay (ELISA), In which either neo-STX-BSA or neo-STXKLH was coated to the mlcroplate, was used to monitor the antibody titer. Although high antibody titers were obtained from rabbits after immunization with both immunogens, only antibody obtained from rabbits immunized with neo-STX-KLH was useful for Immunoassay. Competitive indirect ELISA revealed that the antibodies obtained from rabbits Immunized with neo-STX-KLH are specific for neo-STX but also have good cross-reactivity with STX. The concentrations causing 50% inhibition binding of neo-STX-BSA to the anti-neo-KLH by neo-STX, STX, and decarbamoyl-STX (DC-STX) were 0.9,8.0, and 53.1 ng/mL, respectively. Saxitoxin conjugated to polylyslne (STX-PLL) was also used as the coating reagent In the indirect ELISA. The concentrations causing 50% inhibition binding of antl-neo-STX-KLH to STX-PLL coated on the mlcrotiter plate by neo-STX, STX, and DC-STX were 1.2,4.1, and 36.1 ng/mL, respectively. With this newly developed antibody, ELISA could be a very effective method for monitoring seafood for both neo-STX and STX.

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