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

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.

2013 ◽  
Vol 651 ◽  
pp. 280-283
Author(s):  
Hui Ying Zhang ◽  
Jun Ping Wang

Polyclonal antibody against streptomycin was prepared by using a streptomycin–bovine serum albumin conjugate for the immunization of rabbits. Using this antibody, we developed quantitative assays for streptomycin by means of an indirect competitive enzyme-linked immunosorbent assay (icELISA). Fifty percent inhibition concentration (IC50) for the antibody was 3.6 ng/ml. The detection limit was 0.4 ng/ml. The average of recoveries for all samples was 86.14% and the coefficients of variation of intra- and inter-assays were below 18%. The detection limit using the kit was 15 ng/ml in animal feeds.


1986 ◽  
Vol 49 (4) ◽  
pp. 267-271 ◽  
Author(s):  
RU-DONG WEI ◽  
WILLIAM BISCHOFF ◽  
FUN SUN CHU

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.


2011 ◽  
Vol 1 (2) ◽  
pp. 23-31
Author(s):  
Musyirna Rahmah Nst ◽  
Tri Budhi Murdiati

 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.


1985 ◽  
Vol 31 (10) ◽  
pp. 1625-1631 ◽  
Author(s):  
B Thong ◽  
S J Soldin ◽  
C A Lingwood

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.


1992 ◽  
Vol 75 (2) ◽  
pp. 341-345 ◽  
Author(s):  
Fun S Chu ◽  
Xuan Huang ◽  
Sherwood Hall

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.


2004 ◽  
Vol 22 (SI - Chem. Reactions in Foods V) ◽  
pp. S199-S202 ◽  
Author(s):  
M. Vítková ◽  
Z. Macková ◽  
R. Koblovská ◽  
O. Lapčík

The development of polyclonal antibody-based enzyme-linked immunosorbent assays (ELISA) for the determination of individual isoflavones, i.e. daidzein, genistein and biochanin and their homologues, is presented in this work. Isoflavone conjugates with bovine serum albumin were used as immunogens, coupled at the position C 7 and C 4’ via a carboxy methyl spacer. The developed ELISAs are highly specific, I<sub>50</sub> values of the standard curves range between 0.3–1.2 ng/ml. The cross reactivities to other isoflavones are in acceptable range and the interference of non-isoflavonoid molecules is negligible. The immunoassays have been used for monitoring of changes in isoflavone levels in alimentary important plants, such as Medicago sativa, during germination; and during different vegetation stages of the Rutaceae family plants.


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