摘要:

A small‐volume filtration method was developed for use in the analysis of total, bound and dissolved residues of four agrochemicals which differed in physico‐chemical properties, in turbid water. Low immunoassay recoveries of molinate, chlorpyrifos and endosulfan (but not diuron) in highly turbid water samples suggested that the antibodies were unable to recognize particle‐bound residues. However, in samples of moderate or low turbidity, the results indicated that the data obtained by immunoassay s may approximate the total residue load in samples, especially if the samples are filtered through a non‐absorptive membrane. The adsorption pattern of each membrane was different for each pesticide, and related to their water solubilities and partition coefficients. A syringe filter with an aluminium oxide membrane was more suitable than one with cellulose acetate, polyvinylidene difluoride or polytetrafluoroethylene for filtration of small volumes of water for separation of the dissolved residues, and removal of the residue that desorbed during filtration, without losses of the pesticide on to the membrane. This method should facililate ecotoxicological analysis by enabling the direct analysis of agrochemicals by immunoassay in field water samples.

ISSN:0954-0105    

DOI:10.1080/09540109809354963

出版商:Taylor & Francis Group    

年代:1998

数据来源: Taylor

摘要:

A panel of monoclonal antibodies (MAbs) was raised against tea polyphenols conjugated to bovine serum albumin. A cross‐reactivity study carried out against various purified polyphenols showed that most of the antibodies generated have the ability to recognize epigallocatechin > epicatechin > thearubigin = theaflavin > epicatechin‐3‐gallate = epigallocatechin‐3‐gallate > rutin. One of the selected MAbs, 5756.3, was used in an immunoassay to measure polyphenols in tea, urine and blood samples.

ISSN:0954-0105    

DOI:10.1080/09540109809354964

出版商:Taylor & Francis Group    

年代:1998

数据来源: Taylor

摘要:

An ELISA method to screen the fungicide metalaxyl in wine was developed. To help prevent matrix effects, wine samples were diluted 1: 10 in a solution of sodium bisulfite, dibasic sodium phosphate and polyvinylpyrrolidone. To determine the accuracy and reproducibility of the method, spiked wine samples were employed. Percentage recoveries ranged from 85.3 to 112, while intra‐assay percentage coefficients of variation (%CVs) varied from 6.5 to 16 and inter‐assay %CVs ranged from 10 to 22. A comparison between highperformance liquid chromatography and ELISA was examined utilizing 32 samples. A correlation coefficient of 0.9919 was obtained.

ISSN:0954-0105    

DOI:10.1080/09540109809354965

出版商:Taylor & Francis Group    

年代:1998

数据来源: Taylor

摘要:

Chicken antibodies offer many advantages over the traditional mammalian ones. A laying hen produces large amounts of yolk antibodies and the use of yolk antibodies eliminates the painful procedure of collecting blood from the animal. Thus, the use of chicken antibodies will reduce both the number of animals required to produce antibodies and also animal distress. Chicken antibodies also have several biochemical advantages compared to mammalian antibodies: they often increase the signal and reduce interference in many assays. However, the species chosen for antibody production have usually been mammals. This is probably due to tradition, but also to limited knowledge about the production of chicken antibodies. We studied the immune response in the chicken using small amounts of mammalian antigen, and show that a good immune response can be obtained with 0.1–1.0 μg of bovine serum albumin.

ISSN:0954-0105    

DOI:10.1080/09540109809354966

出版商:Taylor & Francis Group    

年代:1998

数据来源: Taylor

摘要:

Aroclor 1254 (consisting of a mixture of over 100 different congeners of polychlorinated biphenyls (PCBs) was adsorbed on to the surface of polystyrene microbeads and used to immunize mice and rabbits. High titre antisera were produced. PCBs were also adsorbed on to antimony pentoxide microbeads which were then used effectively as immobilized antigen in microbead‐based ELISA assays. Antisera raised with Aroclor 1254‐coated beads contained antibodies with high affinity (as determined by BIAcore analysis) against tetracholoro‐biphenyls even though these congeners represent only a minor fraction in Aroclors.

ISSN:0954-0105    

DOI:10.1080/09540109809354967

出版商:Taylor & Francis Group    

年代:1998

数据来源: Taylor

摘要:

Polyclonal chicken antibodies raised against bovine serum albumin (BSA) were immobilized on chitosan gel for the immunoaffinity isolation of BSA from cow's milk. Antibodies (IgY) against BSA were isolated from egg‐yolk, purified and antibody reactivity to antigen was measured. IgY developed against BSA was reduced by 2‐mercaptoethylamine. The reactivities of reduced and whole IgY against BSA were not significantly different. The reduced IgY was covalently linked to chitosan gel through stable covalent thioether linkages using sulfo‐succinimidyl‐4‐(N‐maleimidomethyl)cyclohexane‐l‐carboxylate (sulfo‐SMCC) as a cross‐linker. The density of antibody IgY immobilized on chitosan gel was approximately 3–5 mg per ml of chitosan gel. The ligand‐binding capacity of immobilized IgY towards BSA was 0.35–0.44 mg BSA per ml of chitosan gel. A single pass of skimmed milk through the column allowed the removal of BSA from the milk sample. The milk sample was analyzed, before and after immunoaffinity separation, by SDS‐PAGE. BSA was desorbed with 0.5 M‐glycine‐HCl buffer at pH 2.8 but the reusability of the column was limited to three cycles. Alternatively, BSA was desorbed with 0.5 M‐glycine‐HCl buffer containing 2 M‐NaCl at pH 4.6 after longer incubation times at a slower flow rate. The low ligand‐binding capacity was not an impedement to reuse of the column. The column was reused more than 20 times with minimal loss of binding capacity.

ISSN:0954-0105    

DOI:10.1080/09540109809354968

出版商:Taylor & Francis Group    

年代:1998

数据来源: Taylor

摘要:

Buffalo ferritin has been isolated and purified from liver using conventional biochemical techniques such as thermal denaturation, ammonium sulphate fractionation, Sephacryl S‐300 gel filtration and DEAE‐blue gel affinity chromatography. Native gel‐electrophoresis of affinity‐purified ferritin followed by iron staining showed a single band corresponding to rat liver ferritin. The yield and the iron content of purified ferritin were 10.7 mg per 500 g of liver and 7% respectively. As a glycoprotein, buffalo ferritin has 12.2% neutral carbohydrate. Polyclonal antibodies raised against purified buffalo liver ferritin in rabbits were used successfully in the development of competitive indirect ELISA using alkaline phosphatase as an enzyme label. The sensitivity range of the assay was 5–30 ng with intraand inter‐assay coefficients of variation (CVs) of 3.1% and 9.2% respectively. The assay was reproducible and could be applied under field conditions for the evaluation of ferritin levels in buffalo. Using this method, the range of circulating ferritin levels in 10 normal buffalo was 1.12–3.68 μg ml‐1. Cross‐reactivity studies by ELISA also suggest the applicability of the method to the evaluation of ferritin levels in other domestic species such as sheep and cattle.

ISSN:0954-0105    

DOI:10.1080/09540109809354969

出版商:Taylor & Francis Group    

年代:1998

数据来源: Taylor

摘要:

Dothistromin (DOTH) is a toxin secreted byDothistroma piniand is present in needle lesions resulting from infection by this fungus. A monoclonal antibody (MAb 10C12) of high affinity for DOTH was conjugated to bovine serum albumin (BSA) and used to immunize mice in order to prepare monoclonal anti‐idiotypic antibody mimics of dothistromin. One anti‐idiotypic antibody, 12C9G8, was equivalent to DOTH on a molar basis in inhibition of the binding of an anti‐DOTH MAb to DOTH‐protein conjugates. Furthermore, in a Western blot analysis, 12C9G8 visualized the same protein profile in embryos as was observed in an approach which utilized an anti‐DOTH MAb together with DOTH‐mouse serum albumin (MSA) conjugate to identify potential DOTH binding sites. In a parallel immunogold electron microscopy experiment, protein‐containing vesicles were labelled using both approaches.

ISSN:0954-0105    

DOI:10.1080/09540109809354970

出版商:Taylor & Francis Group    

年代:1998

数据来源: Taylor

摘要:

In previous papers, we demonstrated that viableLactoacillus caseissp.caseiadministered orally had a good adjuvant activity at the mucosal level and protected againstSalmonella typhimuriuminfection. We also demonstrated that the mixture ofL. caseiplus LPS enhanced its protective capacity without side‐effects. In this paper, we study the frequency and duration of boosters withL. caseinecessary to maintain a good mucosa immune response that protects against infection, without alteration in the immune cells associated with the lamina propia of small intestine. After priming withL. casei,different boosting protocols with single or double doses ofL. caseiwere made. We determined, on the lamina propia of the small intestine, IgA and IgM producing cells, CD4+and CD8+T‐cells and the number of macrophages and polymorphonuclear (PMN) cells. Protective assays againstS. typhimuriuminfection and assays of specific anti‐Salmonella IgA present in the intestinal fluid were also performed. We demonstrate that boosters with only one dose on the 15th or 30th day post‐priming were more effective in protecting againstS. typhimuriuminfection than a booster with two doses ofL. casei,which was not effective. The secretory‐IgA levels, IgA‐secreting cells, macrophages, CD4+, CD8+and T‐cells were increased with the effective boosters. We also demonstrate that the ratio of CD4+/CD8+T‐cell populations was not altered with the different boosting protocols. The conditions for the use ofL.caseissp.caseiCRL 431 as an oral adjuvant or as a vaccine vector were determined.

ISSN:0954-0105    

DOI:10.1080/09540109809354971

出版商:Taylor & Francis Group    

年代:1998

数据来源: Taylor

ISSN:0954-0105    

DOI:10.1080/09540109809354962

出版商:Taylor & Francis Group    

年代:1998

数据来源: Taylor