Effects of hapten structure and bridging groups on antisera specificity in parathion immunoassay development

Vallejo, Remo P.; Bogus, Edward R.; Mumma, Ralph O.

doi:10.1021/jf00111a040

Cited by 61 publications

(32 citation statements)

References 11 publications

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“…First, the presence of a hemisuccinate substituent facilitated the synthesis of protein conjugates by appending a carboxyl-bearing moiety on FIAU. Second, bridging groups are frequently added to small molecules to enhance the formation of hapten-specific antibodies (8,53). Third, protein conjugates of 5'-O-hemisuccinate derivatives have successfully been used to produce rabbit antibodies against the nucleosides 1-0-D-arabinofuranosylcytosine (ara-C) and 1-13-D-arabinofuranosyluracil (33)(34)(35)45).…”

Section: Discussionmentioning

confidence: 99%

Sensitive and specific radioimmunoassay for fialuridine: initial assessment of pharmacokinetics after single oral doses to healthy volunteers

Bowsher

Compton

Kirkwood

et al. 1994

Antimicrob Agents Chemother

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Fialuridine (FIAU) is a halogen-substituted analog of thymidine that was undergoing clinical investigation as a drug for the treatment of chronic hepatitis B viral infection. However, clinical trials of FIAU were terminated after adverse events occurred following chronic oral administration. Prior to the termination of clinical trials, a sensitive assay was needed for the measurement of FIAU because of the anticipated low dose administered to patients. We therefore undertook the development of a radioimmunoassay (RIA). A specific antiserum was raised in rabbits following immunization with a 5'-O-hemisuccinate analog of FIAU coupled to keyhole limpet hemocyanin. Radiolabeled FIAU was synthesized by a destannylation procedure by using sodium [125j]iodide. We developed a competitive-binding procedure and used precipitation with polyethylene glycol as the method for separating the bound and free forms of FIAU. The RIA is sensitive (0.2 ng/ml), specific (negligible interference from known metabolites and endogenous nucleosides), and reproducible (interassay coeflicients of variation range from 5 to 19.7% for serum controls). We used the RIA to assess the pharmacokinetics of FIAU in healthy adult volunteers following administration of a single 5-mg oral dose. The sensitivity of the RIA permitted the detection of a prolonged elimination phase for FIAU in healthy volunteers and dogs, with mean elimination half-lives of 29.3 and 35.3 h, respectively. We conclude the RIA is a valid method for the quantification of FIAU in biological fluids.Fialuridine (FIAU), 1-(2'-deoxy-2'-fluoro-13-D-arabinofuranosyl)-5-iodouracil (Fig. 1, compound 2), is one of a series of 2'-fluoro-substituted arabinosyl pyrimidine nucleosides that have demonstrated potent antiviral activities against a number of clinically important viruses, including hepatitis B virus (HBV) (7,11,28,47). Analogs of FIAU have been shown to inhibit viral replication in the woodchuck and duck models of HBV infection (18,19). Although the mechanism of the anti-HBV activity is not well understood, evidence suggests that the triphosphate analog of FIAU is a potent inhibitor of HBV DNA polymerase activity (17,23,43). During early clinical investigation FIAU showed much promise as an anti-HBV drug because it markedly reduced the level of HBV DNA in the serum of patients with chronic hepatitis B (20). However, clinical trials were terminated after adverse events occurred following oral administration of FIAU (0.1 and 0.25 mg/kg of body weight per day) for more than 2 months (24, 29). The mechanism of the unexpected delayed toxicity is unresolved.Prior to the termination of clinical trials an assay was needed to quantify FIAU in biological fluids. However, the anticipated low therapeutic dose in patients necessitated the development of a highly sensitive analytical method. The criterion of sensitivity precluded the use of an existing UV-based high-pressure liquid chromatographic (HPLC) method for FIAU because its

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Section: Discussionmentioning

confidence: 99%

Sensitive and specific radioimmunoassay for fialuridine: initial assessment of pharmacokinetics after single oral doses to healthy volunteers

Bowsher

Compton

Kirkwood

et al. 1994

Antimicrob Agents Chemother

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“…This method produced a 16-atom linker arm between the ligand (FBi) and the protein. Previous studies have reported positive effects of increased linker arm length on the specificity and the sensitivity of the resulting antibodies (Vallejo et al, 1982;Wei & Hammock, 1984;Harrison et al, 1991;Sheth & Sporns, 1991;KampsHoltzapple et al, 1994). Extending small haptens away from the protein mass generally has been shown to be helpful, or indeed even critical, in the production of high-affinity antibodies (Hastings et al, 1988;Kamps-Holtzapple et al, 1994).…”

Section: Modeling Of Fb! Fb 2 Fb 3 and Hb-fbimentioning

confidence: 98%

Development of an improved monoclonal antibody‐based Elisa for fumonisin b_1–3and the use of molecular modeling to explain observed detection limits

Elissalde

Kamps‐Holtzapple

Beier

et al. 1995

Food and Agricultural Immunology

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1995) Development of an improved monoclonal antibody-based Elisa for fumonisin b 1-3 and the use of molecular modeling to explain observed detection limits, Food and Agricultural Immunology, 7:2, 109-122, Monoclonal antibodies were prepared against the fumonisins, a group of mycotoxins produced by the plant pathogen, Fusarium moniliforme. Splenic lymphocytes, from Balb/c mice immunized with fumonisin B 1 -ovalbumin conjugate, were fused with SP2/O myeloma cells, and 14 hybridomas were selected. In a competitive enzyme-linked immunosorbent assay, fumonisin B 1 -bovine serum albumin and free fumonisin B 1 (FB 1 ) competed for the monoclonal antibody. The concentrations of FB 1 required to inhibit 50% antibody binding (IC 50 ) ranged from 300 to 670 ppb. Antibodies also cross-reacted with fumonisins B 2 and B 3 (FB 2 , FB 3 ), and the hydrolyzed backbone of fumonisin B 1 (HB-FB 1 ). None of the 14 monoclonal antibodies recognized the sphingolipids, sphingosine and sphinganine, that are structurally similar to the backbone of the fumonisins. Three-dimensional computer models of FB 1 , FB 2 and FB 3 show the amine backbone folding with the two esterified trimethylpropane-1,2,3-tricarboxylic acid side-chains to form a cage into which the hydroxyl and acid groups of these fumonisins extend. The HB-FB 1 molecule, with the two trimethylpropane-1,2,3-tricarboxylic acid esterified moieties at carbons 14 and 15 removed, does not possess two of the three branches which are folded together with inter-hydrogen bonding to formulate the three-dimensional structure that makes up the cage feature of FB 1 , FB 2 and FB 3 . Because of the unexpected folding of the three arms to make a cage of FB 1 , attachment of the protein to the amine group, which is close to, or appears to be pari of, the epitope, may have allowed the immune system of the mouse to produce antibodies more specific for the FB 1 -protein conjugate than to free FB 1 .

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“…8, 1992 McAdam et al Direct diazo coupling of the reduced amine form of the aromatic nitro-organophosphate pesticide has been used with moderate success in the case of parathion, which is structurally related to FN (Ercegovich et al, 1981). Subsequent work (Vallejo et al, 1982) used a variety of spacer arms, including two linked through the reduced nitro group. The glutarimine spacer arm used by these workers is similar to the adipic acid spacer used in the present study.…”

Section: Antibody Response To Different Fn-proteinmentioning

confidence: 99%

Mono- and polyclonal antibodies to the organophosphate fenitrothion. 1. Approaches to hapten-protein conjugation

McAdam¹,

Hill²,

Beasley³

et al. 1992

J. Agric. Food Chem.

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Effects of hapten structure and bridging groups on antisera specificity in parathion immunoassay development

Cited by 61 publications

References 11 publications

Sensitive and specific radioimmunoassay for fialuridine: initial assessment of pharmacokinetics after single oral doses to healthy volunteers

Sensitive and specific radioimmunoassay for fialuridine: initial assessment of pharmacokinetics after single oral doses to healthy volunteers

Development of an improved monoclonal antibody‐based Elisa for fumonisin b_1–3and the use of molecular modeling to explain observed detection limits

Mono- and polyclonal antibodies to the organophosphate fenitrothion. 1. Approaches to hapten-protein conjugation

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Effects of hapten structure and bridging groups on antisera specificity in parathion immunoassay development

Cited by 61 publications

References 11 publications

Sensitive and specific radioimmunoassay for fialuridine: initial assessment of pharmacokinetics after single oral doses to healthy volunteers

Sensitive and specific radioimmunoassay for fialuridine: initial assessment of pharmacokinetics after single oral doses to healthy volunteers

Development of an improved monoclonal antibody‐based Elisa for fumonisin b1–3and the use of molecular modeling to explain observed detection limits

Mono- and polyclonal antibodies to the organophosphate fenitrothion. 1. Approaches to hapten-protein conjugation

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Development of an improved monoclonal antibody‐based Elisa for fumonisin b_1–3and the use of molecular modeling to explain observed detection limits