- W. Haasnoot (6)
- W. Keizer de (1)
- F. Kohen (2)
- T. Korpimaeki (1)
- U. Lamminmaeki (1)
- M.E. Ploum (2)
- J.A. Rhijn van (1)
- B. Schacht (1)
- E. Schacht (1)
- M. Swanenburg (1)
Flow cytometric immunoassay for sulfonamides in raw milk
Keizer, W. de; Bienenmann-Ploum, M. ; Bergwerff, A.A. ; Haasnoot, W. - \ 2008
Analytica Chimica Acta 620 (2008)1-2. - ISSN 0003-2670 - p. 142 - 149.
sulfamethazine residues - biosensor immunoassay - immunobiosensor - sulfadiazine - serum
Sulfonamide antibiotics are applied in veterinary medicine for the treatment of microbial infections. For the detection of residues of sulfonamides in milk, a multi-sulfonamide flow cytometric immunoassay (FCI) was developed using the Luminex MultiAnalyte Profiling (xMAP) technology. In this automated FCI, a previously developed biotinylated multi-sulfonamide mutant antibody (M.3.4) was applied in combination with fluorescent beads, directly coated with a sulfathiazole derivative, and streptavidin¿phycoerythrin (SAPE) for the detection. With this FCI, at least 11 different sulfonamides could be detected (more than 50% inhibition at the 100 ng mL¿1 level) and, after an incubation of 1 h, measurements were rapid (10 s per sample). For the application with raw milk, a 96-well microplate-based filtration step was included into the protocol to remove disturbing milk fat particles. Because of differences in sensitivity towards different sulfonamides, the FCI was considered and validated as a qualitative screening assay. For sulfadoxine, the most applied sulfonamide in Dutch dairy cattle, the detection capability (CCß) was
Application of a multi-sulfonamide biosensor immunoassay for the detection of sulfadiazine and sulfamethoxazole residues in broiler serum and its use as a predictor of the levels in edible tissue
Haasnoot, W. ; Ploum, M.E. ; Lamminmaeki, U. ; Swanenburg, M. ; Rhijn, J.A. van - \ 2005
Analytica Chimica Acta 552 (2005)1/2. - ISSN 0003-2670 - p. 87 - 95.
enzyme-immunoassay - sulfamethazine residues - antibody - milk - pharmacokinetics - immunobiosensor - trimethoprim - urine - pigs
A multi-sulfonamide biosensor immunoassay (BIA), based on a previously developed mutant antibody (A.3.5) in an optical biosensor (Biacore 3000), was applied to analyse the serum and plasma samples obtained from the broilers treated with sulfamethoxazole and sulfadiazine. The assay was fast (5 min per sample), the sample preparation easy (dilution in antibody containing buffer only) and an equal sensitivity for the two sulfonamides was obtained with limits of detection (LOD) in serum and plasma below 10 ng ml¿1. The concentrations found with the BIA in serum and plasma of the treated broilers were comparable and higher than the concentrations found in the tissue by LC¿MS/MS. The average serum/tissue ratios for sulfamethoxazole were 6.2 (leg meat), 2.5 (liver) and 1.3 (skin + fat) and for sulfadiazine 8.7 (leg meat), 3.1 (liver) and 2.2 (skin + fat). To predict the concentrations of the two sulfonamides below the maximum residue limit (MRL) of 100 ng g¿1 in the tissue with the highest level (skin + fat), the proposed action level of the multi-sulfonamide BIA in serum is 130 ng ml¿1. A later developed mutant antibody (M.3.4), with a better sensitivity towards more sulfonamides, was applied during a survey. Serum samples (n = 300) of broilers from 30 different flocks were found negative. Concentrations between
Comparison of multi-sulfonamide biosensor immunoassays
Ploum, M.E. ; Korpimaeki, T. ; Haasnoot, W. ; Kohen, F. - \ 2005
Analytica Chimica Acta 529 (2005)1-2. - ISSN 0003-2670 - p. 115 - 122.
enzyme-immunoassay - sulfamethazine residues - antibodies - assay - immunobiosensor - sulfadimidine - sulfadiazine - generation - extraction - tissues
Three different group-specific anti-sulfonamide antibodies were compared in inhibition assay formats in an optical biosensor (BIACORE 3000) using CM5 sensor chips coated with three different sulfonamide derivatives. The antibodies used were an anti-sulfamethazine monoclonal antibody (Mab) 21C7, the sulfonamide binding protein (SBP) in the Qflex Kit Sulfonamides and a recently developed mutant antibody (M.3.4). Each of these antibodies showed interactions with all 17 sulfonamides tested and one (Mab 21C7) was sensitive for the N4-acetyl metabolites also. The limits of detection of the different sulfonamides in chicken serum varied between 7 and >1000 ng ml¿1 (Mab 21C7), 15 and 340 ng ml¿1 (Qflex) and 4 and 82 ng ml¿1 (Mutant M.3.4). The mutant M.3.4 based assay was found to be the most sensitive towards most of the sulfonamides whereas the Qflex Kit Sulfonamides detected the five sulfonamides registered for application in poultry in The Netherlands within the narrowest measurement range
Generation of group-specific antibodies against sulfonamides
Cliquet, P. ; Cox, E. ; Haasnoot, W. ; Schacht, E. ; Goddeeris, B.M. - \ 2003
Journal of Agricultural and Food Chemistry 51 (2003)20. - ISSN 0021-8561 - p. 5835 - 5842.
enzyme-immunoassay - monoclonal-antibodies - sulfamethazine residues - sulfathiazole - elisa - tissue - recognition - antibiotics - milk - meat
To develop a sulfonamide-specific ELISA, different attempts were made to obtain monoclonal antibodies specific for the common structure of sulfonamides. In a first approach, sulfanilamide was linked to albumins using glutaraldehyde or a succinimide ester as cross-linker. A weak immune response or none at all was induced after immunization of mice with those conjugates. High antibody titers were obtained with conjugates where sulfanilamide was linked to albumins or casein (azocasein) with a diazotation reaction. However, the antibodies were only highly specific for the bound sulfanilamide molecule. In a second approach, sulfonamide-protein conjugates were used in which the sulfonamide molecule is linked at its side chain, leaving the common structure of sulfonamides unchanged. Three sulfonamide derivatives (S, TS, and PS, previously described in the literature) containing a carboxyl group in their side chain were linked to proteins using a carbodiimide mediated reaction. Immunization with the S-conjugates led to high antibody titers, but the antibodies were only highly specific for the bound S-molecule. Group-specific antibodies were obtained after immunization with the PS- and TS-conjugates. It was described that immunization with PS-conjugates lead to the recognition of other sulfonamides (sulfamethazine, -merazine, -diazine, and -dimethoxine) that are not well recognized by antibodies induced after immunization with TS-conjugates. Therefore, we tried to guide the immune response in the direction of recognition of the common structure of sulfonamides by immunizing the animals alternately with PS- and TS-conjugates. The polyclonal antibodies of the mice indeed had a broader specificity, but the specificity of the monoclonals obtained after fusion experiments was not influenced. Immunization with TS-conjugates seemed sufficient to obtain sulfonamide-specific monoclonal antibodies. With the best monoclonal (mAb 3B5B10E3) two competitive inhibition (ci) ELISA's were developed: one coated with antigen and the other coated with the monoclonal antibody. Sulfadiazine, -dimethoxine, -thiazole, -pyridine, and -methoxazole were detected in both ELISA's at their MRL-value (100 ppb) in buffer solution. Sulfadiazine, sulfathiazole, and sulfamethoxazole could even be detected at 10 ppb.
Extraction procedure for sulfachloropyridazine in porcine tissues and detection in a sulfonamide-specific enzyme-linked immunosorbent assay (ELISA)
Cliquet, P. ; Cox, E. ; Haasnoot, W. ; Schacht, B. ; Goddeeris, B.M. - \ 2003
Analytica Chimica Acta 494 (2003)1-2. - ISSN 0003-2670 - p. 21 - 28.
sulfamethazine residues - monoclonal-antibodies - animal-tissues - swine urine - immunoassay - sulfathiazole - sulfadimidine - antibiotics - meat - recognition
Sulfonamide-specific polyclonal rabbit antibodies were obtained after immunization with a sulfathiazole derivative (N1-[4-(carboxymethyl)-2-thiazolyl]sulfanilamide = TS) coupled to keyhole lympet hemocyanin. Using these antibodies, two sulfonamide-specific enzyme-linked immunosorbent assays (ELISAs) were developed differing in coating antigen: TS-ovalbumin (TS-ova) and PS-ovalbumin (PS-ova, PS = N1-[4-methyl-5-[2-4-carboxyethyl-1-hydroxyphenyl]-azo-2-pyridyl]sulfanilamide). The detection of sulfamethazine, sulfamerazine, sulfadimethoxine, sulfadiazine, sulfathiazole, sulfapyridine, sulfachloropyridazine and sulfisoxazole in buffer was studied. Higher antibody titers were obtained in the ELISA coated with TS-ova (TS-ciELISA) as compared to the ELISA coated with PS-ova (PS-ciELISA), but the detection of sulfonamides was more sensitive in the PS-ciELISA, allowing the detection of all tested sulfonamides at the maximum residue level (MRL) value (100 ng ml¿1). In a subsequent step, an extraction procedure was developed for the detection of sulfonamides in muscles, kidney, liver and fat by both ELISAs using sulfachloropyridazine as a model. As extraction buffer a carbonate/hydrogen carbonate buffer (pH 10) was chosen in which sulfonamides are highly soluble. Differences in homogenizing techniques (high-speed mixer (Ultraturax) versus vortex) and the effect of kaolin (hydrated aluminum silicate) treatment, to diminish the background signal in the ELISA, were evaluated. The best extraction procedure was the one using a vortex mixer as homogenizer and no kaolin treatment. Sulfachloropyridazine was easily detected at the MRL in all tissues.
Biosensor immunoassay for the detection of eight sulfonamides in chicken serum
Haasnoot, W. ; Bienenmann-Ploum, M. ; Kohen, F. - \ 2003
Analytica Chimica Acta 483 (2003)1-2. - ISSN 0003-2670 - p. 171 - 180.
enzyme-immunoassay - sulfamethazine residues - 2 sulfonamides - antibodies - assay - immunobiosensor - sulfadiazine - extraction - tissues - kidney
A monoclonal antibody (MAb) raised against sulfamethazine (21C7) was applied in an optical biosensor (Biacore Q) to develop a rapid biosensor immunoassay (BIA) for the detection of several sulfonamides in chicken serum. The performance of this MAb was compared with two polyclonal antibodies (PAbs) raised against sulfamethazine (Qflex sulfamethazine binding protein (SBP) and RIKILT 464b). Using these PAbs, the limits of detection (LODs) in 10 times diluted chicken serum were approximately 30 ng ml-1 and the two BIAs were found to be specific for sulfamethazine. Using MAb 21C7, the LOD for sulfamethazine in 10 times diluted chicken serum was lower (10 ng ml-1), high cross-reactivities were measured for sulfisoxazole (149¿ sulfachlorpyridazine (112¿ sulfachlorpyrazine (94¿ sulfamerazine (87¿ sulfadiazine (56¿ sulfatroxazole (56¿and sulfathiazole (50¿and low cross-reactivities (11¿25¿were measured with six other sulfonamides. Compared with the polyclonal antibodies, the MAb-based BIA resulted in a better sensitivity and was found suitable for the detection of 8 sulfonamides in 10 times diluted chicken serum with LODs between 7 and 20 ng ml-1. The total run time for each cycle was 7 min.