Production and Characterization of Antibodies Against Neosaxitoxin

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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Production and Characterization of Antibody Against Deoxynivalenol Triacetate

Journal of Food Protection ◽

10.4315/0362-028x-49.5.336 ◽

1986 ◽

Vol 49 (5) ◽

pp. 336-339 ◽

Cited By ~ 14

Author(s):

GUANG-SHI ZHANG ◽

SHU WEN LI ◽

FUN SUN CHU

Keyword(s):

Bovine Serum Albumin ◽

Serum Albumin ◽

Bovine Serum ◽

Detection Limits ◽

Cross Reactivity ◽

Booster Injection ◽

Antibody Titers

Antibodies against deoxynivalenol-triacetate (Tri-Ac-DON) was prepared by immunizing rabbits with a hemisuccinate derivative of 7,8 dihydroxycalonectrin (DHC) conjugated to bovine serum albumin. Using tritiated Tri-Ac-DON as the testing ligand, antibody titers were observed as early as 4 wk after immunization. Useful antibody for radioimmunoassay (RIA) of Tri-Ac-DON was obtained from the rabbits 7 wk after immunization, with one booster injection. Competitive RIA revealed that the antibody was most specific for Tri-Ac-DON. The relative cross-reactivity of this antibody with Tri-Ac-DON, T-2 toxin tetra-acetate, 15-acetyl-deoxynivalenol and acetyl-T-2 toxin was 1 (most cross-reactive), 0.003, 0.002, and 0.001, respectively. Practically no cross-reactivity was observed with deoxynivalenol (DON), diacetoxyscirpenol, nivalenol and T-2 toxin. The detection limits for Tri-Ac-DON by RIA was about 0.1 ng/assay. The use of this antibody for quantitation and confirmation of DON in cereals after acetylation of sample extracts is proposed.

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

Journal of Food Protection ◽

10.4315/0362-028x-49.4.267 ◽

1986 ◽

Vol 49 (4) ◽

pp. 267-271 ◽

Cited By ~ 7

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.

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Easy Enzyme-Linked Immunosorbent Assay for Spectinomycin in Chicken Plasma

Journal of AOAC International ◽

10.1093/jaoac/79.2.426 ◽

1996 ◽

Vol 79 (2) ◽

pp. 426-430 ◽

Cited By ~ 4

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.

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Preparation and characterization of ultrasensitive and specific polyclonal antiserum against ciprofloxacin based on cationized bovine serum albumin

Chemical Papers ◽

10.2478/s11696-014-0593-z ◽

2014 ◽

Vol 68 (11) ◽

Cited By ~ 2

Author(s):

Sheng-Liang Deng ◽

Ping Li ◽

Hong-Bin Liu ◽

Shu-Ming Yang

Keyword(s):

Bovine Serum Albumin ◽

Serum Albumin ◽

Bovine Serum ◽

Limit Of Detection ◽

Polyclonal Antiserum ◽

Residue Level ◽

Enzyme Linked Immunosorbent Assay ◽

Maximum Residue Level ◽

The Eu

AbstractAn ultrasensitive polyclonal antiserum against ciprofloxacin (CPFX) was developed by introducing cationized bovine serum albumin (BSA) as the carrier. Conjugates used as immunogens were synthesized using CPFX coupled to BSA cationized with ethylenediamine and hexamethylenediamine, namely CPFX-eBSA and CPFX-hBSA. The results showed that the antisera immunized with CPFX-hBSA exhibit an improved sensitivity and specificity compared with those immunized with CPFX-eBSA. Under the conditions of optimal heterologous enzyme-linked immunosorbent assay, the optimal antiserum yielded an IC50 of 0.097 ng mL−1, which proves it to be more sensitive than any CPFX antibody. Cross-reactivities with norfloxacin, enrofloxacin, and ofloxacin were < 3.0 %. No cross-reactivities with penicillin and gentamicin were found. The limit of detection was 0.01 ng mL−1, which is far below the maximum residue level established by the EU regulation, suggesting the great potential of the presented antiserum for quantitative assays of CPFX in samples.

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Bovine serum albumin as an immunogenic carrier facilitating the development of hapten-specific monoclonal antibodies

10.1101/2020.11.25.397455 ◽

2020 ◽

Author(s):

Ruotong Zhao ◽

Mingjun Jiang

Keyword(s):

Monoclonal Antibodies ◽

Bovine Serum Albumin ◽

Serum Albumin ◽

Bovine Serum ◽

Limit Of Detection ◽

Fetal Bovine Serum ◽

Cross Reactivity ◽

Enzyme Linked Immunosorbent Assay ◽

Common Misconception ◽

Fetal Bovine

AbstractThere is a common misconception that the generation of hapten-specific monoclonal antibodies (mAbs) requires the use of a heterologous conjugate to ensure carrier-specific antibodies not being detected. In this study, salbutamol (SAL) was used as a model hapten to exhibit the benefits of bovine serum albumin (BSA) as a carrier for developing hapten-specific mAbs. SAL-BSA conjugate would serve as both an immune antigen and a screening antigen during the preparation of SAL-specific mAbs. Six hybridomas were identified to secret mAbs specific for free SAL with minor or negligible cross-reactivity with other β-agonists. Meanwhile, none of hybrodomas secreting anti-BSA antibodies were screened out even though the fetal bovine serum (FBS) added to the medium decreased from 10% to 1% (v/v). Based on one of the six mAbs, 3F12, a direct competitive enzyme-linked immunosorbent assay (dcELISA) was developed for meausring SAL. Under the optimized assay, the quantitative working range was from 312.5 to 20,000 pg/mL (R2 = 0.9959), with a limit of detection (LOD) of 142.9 pg/mL. The results showed that BSA is an efficient and suitable protein carrier for facilitating the development of hapten-specific mAbs.

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