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.

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.

Tetra-sulfonate phthalocyanine zinc-bovine serum albumin conjugate-mediated photodynamic therapy of human glioma

2014 ◽  
Vol 29 (3) ◽  
pp. 378-385 ◽  
Author(s):  
Dianshuang Xu ◽  
Xiangyu Chen ◽  
Ke’en Chen ◽  
Yiru Peng ◽  
Yingxin Li ◽  
...  
Keyword(s):  
Photodynamic Therapy ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Human Glioma ◽  
Bovine Serum Albumin Conjugate

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 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.

PLASMA PROGESTERONE LEVELS IN GUINEA-PIGS ACTIVELY IMMUNIZED AGAINST PROSTAGLANDIN F2α, HYSTERECTOMIZED OR TREATED WITH INTRA-UTERINE INDOMETHACIN

1975 ◽  
Vol 67 (1) ◽  
pp. 81-88 ◽  
Author(s):  
N. L. POYSER ◽  
E. W. HORTON
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Guinea Pigs ◽  
Bovine Serum ◽  
Prostaglandin F2α ◽  
Corpora Lutea ◽  
Plasma Progesterone ◽  
Progesterone Secretion ◽  
Bovine Serum Albumin Conjugate ◽  
Prostaglandin F

SUMMARY Five guinea-pigs actively immunized against a prostaglandin F2α(PGF2α)–bovine serum albumin conjugate showed elongated oestrous cycles. During these, corpora lutea were maintained in a functional secretory state as indicated by plasma progesterone levels. The results are compatible with the view that the PGF2α antibodies neutralized the PGF2α released from the uterus and thus prevented its normal luteolytic effect. Similar patterns of progesterone secretion were observed in two hysterectomized animals and in two animals with intra-uterine implants of indomethacin.

Screening radioimmunoassay for aldosterone in preheated plasma without extraction and chromatography.

1980 ◽  
Vol 26 (1) ◽  
pp. 41-45
Author(s):  
T M Connolly ◽  
L Tibor ◽  
K H Gless ◽  
P Vecsei
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Binding Proteins ◽  
Equilibrium Dialysis ◽  
Screening Method ◽  
Plasma Aldosterone ◽  
Plasma Samples ◽  
Bovine Serum Albumin Conjugate ◽  
Native Plasma

Abstract We directly estimated plasma aldosterone radioimmunologically with use of an antiserum raised against an aldosterone-3-oxime/bovine serum albumin conjugate, the estimation being on samples with and without heating (60 degrees C), and diluted and undiluted. Values so obtained were compared with those by radioimmunoassay after extraction and chromatography. The correlation--even negative values were obtained--was poorest when the steroid was directly estimated in nonheated, undiluted plasma. Correlations were best (r = 0.918) for preheated and diluted native plasma, and the interassay CV was 9.8% (n = 57). However, there were some extraordinarily high values. After equilibrium dialysis of native and preheated (60 degrees C) plasma (15 plasma samples), the percentages of apparent free aldosterone and cortisol increased from 51.4 +/- 2.6% (SEM) to 64.3 +/- 1.6% and from 11.5 +/- 2.2% to 61.1 +/- 1%, respectively. We conclude that aldosterone-binding proteins play a role in direct radioimmunoassays of aldosterone in plasma, but by heating (with or without diluting) the plasma, direct assay can be used as a simple, fast, and inexpensive screening method.

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.

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