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Bruce D. Hammock - One of the best experts on this subject based on the ideXlab platform.
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immunoassay for monitoring environmental and human exposure to the polybrominated diphenyl ether bde 47
Environmental Science & Technology, 2009Co-Authors: Hsing Ju Tsai, Deborah H Bennett, Marcia Nishioka, Arlene Blum, Elana Fishman, Bruce D. HammockAbstract:We developed a selective competitive enzyme-linked immunosorbent assay (ELISA) to monitor environmental and human exposure to polybrominated diphenyl ether BDE-47 that is used as a flame retardant. 2,2′,4,4′-Tetrabromodiphenyl ether (BDE-47), a dominant PBDE congener of toxicological concern, was the target analyte. To achieve effective hapten presentation on the carrier protein for antibody production, immunizing Haptens with a rigid double-bonded hydrocarbon linker introduced at different positions on the target molecule were synthesized as well as coating Haptens that mimic a characteristic fragment of the molecule. Rabbit antisera produced against each immunizing antigen were screened against competitive hapten coating antigens. Under optimized competitive indirect ELISA conditions, the linear detection range in the assay buffer that includes 50% dimethyl sulfoxide was 0.35−8.50 μg/L with an IC50 value of 1.75 μg/L for BDE-47. Little or no cross-reactivity (<6%) was observed to related PBDE congeners ...
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phage borne peptidomimetics accelerate the development of polyclonal antibody based heterologous immunoassays for the detection of pesticide metabolites
Environmental Science & Technology, 2008Co-Authors: Hee Joo Kim, Ki Chang Ahn, Shirley J Gee, Andres Gonzaleztechera, Gualberto Gonzalezsapienza, Bruce D. HammockAbstract:Competitive immunoassays for the detection of small analytes, such as pesticides and their metabolites, use Haptens that compete with the target compounds for binding to the antibody. This competing hapten can be either the same as the immunizing hapten (homologous assay) or structurally modified mimics of the immunizing hapten (heterologous assay). Polyclonal antibody-based heterologous immunoassays have shown superior sensitivities to homologous ones, but the synthesis of heterologous Haptens may be time-consuming, requiring expertise in synthetic chemistry. In this work we demonstrate that phage display peptide libraries can be used as a source of phage-borne peptidomimetics to facilitate the development of sensitive heterologous assays. Different strategies for the isolation of these peptides were explored using two metabolites of pyrethroid insecticides. The sensitivities of the best competitive phage heterologous enzyme-linked immunosorbent assays were 13 fold and 100 fold better than the homologous...
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hapten and antibody production for a sensitive immunoassay determining a human urinary metabolite of the pyrethroid insecticide permethrin
Journal of Agricultural and Food Chemistry, 2004Co-Authors: Takaho Watanabe, Bruce D. HammockAbstract:Permethrin is the most popular synthetic pyrethroid insecticide in agriculture and public health. For the development of the enzyme-linked immunosorbent assay (ELISA) to evaluate human exposure to permethrin, the glycine conjugate (DCCA-glycine) of a major metabolite, cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acid (DCCA), of permethrin was established as the target analyte. Four different types of the cis- and trans-isomers of immunizing Haptens were synthesized as follows: N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine (hapten 3), N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)-4-amino-l-phenylalanine (hapten 5), N-(N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine)amino-6-(2,4-dinitrophenyl)aminohexanoic acid (hapten 9), and N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine-4-oxobutanoic acid (hapten 24). Sixteen polyclonal antibodies produced against each cis- o...
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Synthesis of Haptens for immunoassay of organophosphorus pesticides and effect of heterology in hapten spacer arm length on immunoassay sensitivity
Analytica Chimica Acta, 2003Co-Authors: Yoo Jung Kim, Yong Tae Lee, Young Ae Cho, Hyesung Lee, Shirley J Gee, Bruce D. HammockAbstract:The synthetic method for Haptens of organophosphorus (OP) pesticides with a spacer arm (amino carboxylic acid) attached at the pesticide thiophosphate group was simplified to a large extent. While the previous synthetic approach for this type of Haptens requires seven steps, the present process involves only two steps. Using this process, five Haptens of fenthion differing in spacer arm length (4–8 atoms) were synthesized and they were conjugated to bovine serum albumin and keyhole limpet hemocyanin to be used as immunogens. Rabbits were immunized with these hapten–protein conjugates for production of polyclonal antibodies against fenthion. The five Haptens were conjugated to ovalbumin to be used as plate-coating antigens and twenty polyclonal antisera to the Haptens were screened against each of the five coating antigens using noncompetitive and competitive indirect enzyme-linked immunosorbent assay (ELISA). The titer difference between the homologous and heterologous combinations was small, suggesting that heterology in spacer arm length is not important for the antigen recognition by antibodies. While the heterology in spacer length of the coating antigen had no significant effect on the sensitivity of ELISA, heterology in spacer structure of the coating antigen produced a remarkable improvement in the sensitivity of ELISA. © 2002 Elsevier Science B.V. All rights reserved.
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Synthesis of Haptens and protein conjugates for the development of immunoassays for the insect growth regulator fenoxycarb
Journal of agricultural and food chemistry, 2002Co-Authors: Ferenc Szurdoki, András Székács, Bruce D. HammockAbstract:Sensitive and selective enzyme-linked immunosorbent assays (ELISAs) in the immobilized antigen format were developed for fenoxycarb (1), an insect growth regulator (IGR). The parent molecule [ethyl 2-(4-phenoxyphenoxy)ethylcarbamate] was derivatized at several positions to obtain Haptens (2−5) that were used to produce protein conjugates and rabbit polyclonal antisera. Amino derivatives of fenoxycarb at the terminal and internal rings (2 and 3, respectively) were linked to carrier proteins by azo coupling. Carboxyalkyl-spacer groups were attached to the ethyl group and the nitrogen atom of the target compound (1) to obtain Haptens 4 and 5, respectively. Hapten-homologous ELISAs based on protein conjugates of compounds 2 and 4 determined fenoxycarb in the mid-ppb range (IC50, 102 and 95 ppb, respectively). A more sensitive hapten-heterologous ELISA (IC50, 17 ppb; detection limit 0.5 ppb) involved the antiserum raised against a conjugate of hapten 2 and the plate-coating antigen obtained from compound 3. Th...
Hongtao Lei - One of the best experts on this subject based on the ideXlab platform.
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Hapten synthesis and production of specific antibody against 3-amino-5-morpholinomethyl-2-oxazolidone for immunoassay without derivatisation
'Informa UK Limited', 2018Co-Authors: Fengyin Liu, Yudong Shen, Yuanming Sun, Jinyi Yang, Hong Wang, Zijian Chen, Hongtao LeiAbstract:Immunoassay direct for 3-amino-5-morpholinomethyl-2-oxazolidone (AMOZ), the metabolite of furaltadone, based on a specific antibody was usually unavailable due to its small molecule weight. In this work, a hapten with a double bond and an active carboxyl group was designed and then conjugated to peptide dendrimer to obtain novel multiple Haptens. The Haptens and multiple Haptens were coupled to bovine serum albumin as immunogens. Polyclonal antibodies showed binding to free AMOZ in a heterologous competitive indirect enzyme-linked immunosorbent assay were obtained. The assay showed IC50 (half maximal inhibitory concentration) of 4.1 μg/kg and limit of detection of 0.2 μg/kg for AMOZ. Recoveries of AMOZ from grass carp were tested from 83.1% to 117.0%, with the coefficient of variation below 12%. As no laborious derivatisation procedure was employed, the proposed assay should be useful for the screening of AMOZ for a large number of samples
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novel hapten synthesis for antibody production and development of an enzyme linked immunosorbent assay for determination of furaltadone metabolite 3 amino 5 morpholinomethyl 2 oxazolidinone amoz
Talanta, 2013Co-Authors: Yudong Shen, Yuanming Sun, Katrina Campbell, Yuanxin Tian, Shiwei Zhang, Hongtao Lei, Yueming JiangAbstract:A heterologous competitive indirect enzyme-linked immunosorbent assay (ciELISA) for the determination of the furaltadone metabolite 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) was developed. AMOZ was derivatised with 2-(4-formylphenoxy) acetic acid or 2-(3-formylphenoxy) acetic acid to obtain two novel immunizing Haptens. The ability of these Haptens in producing specific polyclonal antibodies against the nitrophenyl derivative of AMOZ (NPAMOZ) was compared with that of traditional immunizing Haptens (derivatised AMOZ with 3-carboxybenzaldehyle or 4-carboxybenzaldehyle). The results indicated that the novel immunizing Haptens were able to produce antibodies with almost a two-fold improvement in sensitivity of the ciELISA for NPAMOZ in comparison with the existing antibody based ELISAs. The differences in sensitivity were explained by the molecular modeling of the lowest energy conformations of NPAMOZ and the Haptens. Another novel hapten, derivatised AMOZ with 2-oxoacetic acid, was synthesized and used as a heterologous coating hapten. The results showed that this strategy of using only a partial structure of the target molecule as the coating hapten was able to obtain a two to three-fold improvement in sensitivity. This study provided a modern approach for the development of an immunoassay with improved sensitivity for the metabolites of nitrofuran antibiotics.
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broad specificity immunoassay for o o diethyl organophosphorus pesticides application of molecular modeling to improve assay sensitivity and study antibody recognition
Analytical Chemistry, 2010Co-Authors: Yudong Shen, Hongtao Lei, Jinyi Yang, Hong Wang, Wenxu Zheng, Ross C Beier, Guimian Xie, Jiexian Dong, Zhigang She, Yuanming SunAbstract:A monoclonal antibody (mAb) against 4-(diethoxyphosphorothioyloxy)benzoic acid (hapten 1) was raised and used to develop a broad-specificity competitive indirect enzyme-linked immunosorbent assay (ciELISA) for 14 O,O-diethyl organophosphorus pesticides (OPs). Computer-assisted molecular modeling was used to model two-dimensional (2D) and three-dimensional (3D) quantitative structure−activity relationships (QSARs) to study antibody recognition. On the basis of insights obtained from the QSAR models, two heterologous coating Haptens, 4-(diethoxyphosphorothioylamino)butanoic acid (hapten 2) and 4-(diethoxyphosphorothioyloxy)-2-methylbenzoic acid (hapten 3) were designed, synthesized, and used to develop heterologous ciELISAs with significantly improved sensitivity. The heterologous ciELISA using hapten 2 as the coating hapten showed good sensitivity in a broad-specific manner for eight O,O-diethyl OPs and may be used as a screening method for the determination of these OPs. Our studies demonstrated that mole...
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hapten synthesis and antibody production for the development of a melamine immunoassay
Analytica Chimica Acta, 2010Co-Authors: Hongtao Lei, Yudong Shen, Yuanming Sun, Lijun Song, Jinyi Yang, Olivier P Chevallier, Simon A Haughey, Hong Wang, Christopher T ElliottAbstract:Abstract The incorporation of melamine into food products is banned but its misuse has been widely reported in both animal feeds and food. The development of a rapid screening immunoassay for monitoring of the substance is an urgent requirement. Two Haptens of melamine were synthesized by introducing spacer arms of different lengths and structures on the triazine ring of the analyte molecular structure. 6-Aminocaproic acid and 3-mercaptopropionic acid were reacted with 2-chloro-4,6-diamino-1,3,5-triazine (CAAT) to produce hapten 1 [3-(4,6-diamino-1,6-dihydro-1,3,5-triazin-2-ylamino) hexanoic acid] and hapten 2 [3-(4,6-diamino-1,6-dihydro-1,3,5-triazin-2-ylthio) propanoic acid], respectively. The molecular structures of the two Haptens were identified by 1 H nuclear magnetic resonance spectrometry, mass spectrometry and infrared spectrometry. An immunogen was prepared by coupling hapten 1 to bovine serum albumin (BSA). Two plate coating antigens were prepared by coupling both Haptens to egg ovalbumin (OVA). A competitive indirect enzyme-linked immunosorbent assay (ciELISA) was developed to evaluate homogeneous and heterogeneous assay formats. The results showed that polyclonal antibodies with high titers were obtained, and the heterogeneous immunoassay format demonstrated a better performance with an IC 50 of 70.6 ng mL −1 , a LOD of 2.6 ng mL −1 and a LOQ of 7.6 ng mL −1 . Except for cyromazine, no obvious cross-reactivity to common compounds was found. The data showed that the hapten synthesis was successful and the resultant antisera could be used in an immunoassay for the rapid and sensitive detection of this banned chemical.
Yudong Shen - One of the best experts on this subject based on the ideXlab platform.
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Hapten synthesis and production of specific antibody against 3-amino-5-morpholinomethyl-2-oxazolidone for immunoassay without derivatisation
'Informa UK Limited', 2018Co-Authors: Fengyin Liu, Yudong Shen, Yuanming Sun, Jinyi Yang, Hong Wang, Zijian Chen, Hongtao LeiAbstract:Immunoassay direct for 3-amino-5-morpholinomethyl-2-oxazolidone (AMOZ), the metabolite of furaltadone, based on a specific antibody was usually unavailable due to its small molecule weight. In this work, a hapten with a double bond and an active carboxyl group was designed and then conjugated to peptide dendrimer to obtain novel multiple Haptens. The Haptens and multiple Haptens were coupled to bovine serum albumin as immunogens. Polyclonal antibodies showed binding to free AMOZ in a heterologous competitive indirect enzyme-linked immunosorbent assay were obtained. The assay showed IC50 (half maximal inhibitory concentration) of 4.1 μg/kg and limit of detection of 0.2 μg/kg for AMOZ. Recoveries of AMOZ from grass carp were tested from 83.1% to 117.0%, with the coefficient of variation below 12%. As no laborious derivatisation procedure was employed, the proposed assay should be useful for the screening of AMOZ for a large number of samples
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novel hapten synthesis for antibody production and development of an enzyme linked immunosorbent assay for determination of furaltadone metabolite 3 amino 5 morpholinomethyl 2 oxazolidinone amoz
Talanta, 2013Co-Authors: Yudong Shen, Yuanming Sun, Katrina Campbell, Yuanxin Tian, Shiwei Zhang, Hongtao Lei, Yueming JiangAbstract:A heterologous competitive indirect enzyme-linked immunosorbent assay (ciELISA) for the determination of the furaltadone metabolite 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) was developed. AMOZ was derivatised with 2-(4-formylphenoxy) acetic acid or 2-(3-formylphenoxy) acetic acid to obtain two novel immunizing Haptens. The ability of these Haptens in producing specific polyclonal antibodies against the nitrophenyl derivative of AMOZ (NPAMOZ) was compared with that of traditional immunizing Haptens (derivatised AMOZ with 3-carboxybenzaldehyle or 4-carboxybenzaldehyle). The results indicated that the novel immunizing Haptens were able to produce antibodies with almost a two-fold improvement in sensitivity of the ciELISA for NPAMOZ in comparison with the existing antibody based ELISAs. The differences in sensitivity were explained by the molecular modeling of the lowest energy conformations of NPAMOZ and the Haptens. Another novel hapten, derivatised AMOZ with 2-oxoacetic acid, was synthesized and used as a heterologous coating hapten. The results showed that this strategy of using only a partial structure of the target molecule as the coating hapten was able to obtain a two to three-fold improvement in sensitivity. This study provided a modern approach for the development of an immunoassay with improved sensitivity for the metabolites of nitrofuran antibiotics.
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broad specificity immunoassay for o o diethyl organophosphorus pesticides application of molecular modeling to improve assay sensitivity and study antibody recognition
Analytical Chemistry, 2010Co-Authors: Yudong Shen, Hongtao Lei, Jinyi Yang, Hong Wang, Wenxu Zheng, Ross C Beier, Guimian Xie, Jiexian Dong, Zhigang She, Yuanming SunAbstract:A monoclonal antibody (mAb) against 4-(diethoxyphosphorothioyloxy)benzoic acid (hapten 1) was raised and used to develop a broad-specificity competitive indirect enzyme-linked immunosorbent assay (ciELISA) for 14 O,O-diethyl organophosphorus pesticides (OPs). Computer-assisted molecular modeling was used to model two-dimensional (2D) and three-dimensional (3D) quantitative structure−activity relationships (QSARs) to study antibody recognition. On the basis of insights obtained from the QSAR models, two heterologous coating Haptens, 4-(diethoxyphosphorothioylamino)butanoic acid (hapten 2) and 4-(diethoxyphosphorothioyloxy)-2-methylbenzoic acid (hapten 3) were designed, synthesized, and used to develop heterologous ciELISAs with significantly improved sensitivity. The heterologous ciELISA using hapten 2 as the coating hapten showed good sensitivity in a broad-specific manner for eight O,O-diethyl OPs and may be used as a screening method for the determination of these OPs. Our studies demonstrated that mole...
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hapten synthesis and antibody production for the development of a melamine immunoassay
Analytica Chimica Acta, 2010Co-Authors: Hongtao Lei, Yudong Shen, Yuanming Sun, Lijun Song, Jinyi Yang, Olivier P Chevallier, Simon A Haughey, Hong Wang, Christopher T ElliottAbstract:Abstract The incorporation of melamine into food products is banned but its misuse has been widely reported in both animal feeds and food. The development of a rapid screening immunoassay for monitoring of the substance is an urgent requirement. Two Haptens of melamine were synthesized by introducing spacer arms of different lengths and structures on the triazine ring of the analyte molecular structure. 6-Aminocaproic acid and 3-mercaptopropionic acid were reacted with 2-chloro-4,6-diamino-1,3,5-triazine (CAAT) to produce hapten 1 [3-(4,6-diamino-1,6-dihydro-1,3,5-triazin-2-ylamino) hexanoic acid] and hapten 2 [3-(4,6-diamino-1,6-dihydro-1,3,5-triazin-2-ylthio) propanoic acid], respectively. The molecular structures of the two Haptens were identified by 1 H nuclear magnetic resonance spectrometry, mass spectrometry and infrared spectrometry. An immunogen was prepared by coupling hapten 1 to bovine serum albumin (BSA). Two plate coating antigens were prepared by coupling both Haptens to egg ovalbumin (OVA). A competitive indirect enzyme-linked immunosorbent assay (ciELISA) was developed to evaluate homogeneous and heterogeneous assay formats. The results showed that polyclonal antibodies with high titers were obtained, and the heterogeneous immunoassay format demonstrated a better performance with an IC 50 of 70.6 ng mL −1 , a LOD of 2.6 ng mL −1 and a LOQ of 7.6 ng mL −1 . Except for cyromazine, no obvious cross-reactivity to common compounds was found. The data showed that the hapten synthesis was successful and the resultant antisera could be used in an immunoassay for the rapid and sensitive detection of this banned chemical.
Yuanming Sun - One of the best experts on this subject based on the ideXlab platform.
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Hapten synthesis and production of specific antibody against 3-amino-5-morpholinomethyl-2-oxazolidone for immunoassay without derivatisation
'Informa UK Limited', 2018Co-Authors: Fengyin Liu, Yudong Shen, Yuanming Sun, Jinyi Yang, Hong Wang, Zijian Chen, Hongtao LeiAbstract:Immunoassay direct for 3-amino-5-morpholinomethyl-2-oxazolidone (AMOZ), the metabolite of furaltadone, based on a specific antibody was usually unavailable due to its small molecule weight. In this work, a hapten with a double bond and an active carboxyl group was designed and then conjugated to peptide dendrimer to obtain novel multiple Haptens. The Haptens and multiple Haptens were coupled to bovine serum albumin as immunogens. Polyclonal antibodies showed binding to free AMOZ in a heterologous competitive indirect enzyme-linked immunosorbent assay were obtained. The assay showed IC50 (half maximal inhibitory concentration) of 4.1 μg/kg and limit of detection of 0.2 μg/kg for AMOZ. Recoveries of AMOZ from grass carp were tested from 83.1% to 117.0%, with the coefficient of variation below 12%. As no laborious derivatisation procedure was employed, the proposed assay should be useful for the screening of AMOZ for a large number of samples
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novel hapten synthesis for antibody production and development of an enzyme linked immunosorbent assay for determination of furaltadone metabolite 3 amino 5 morpholinomethyl 2 oxazolidinone amoz
Talanta, 2013Co-Authors: Yudong Shen, Yuanming Sun, Katrina Campbell, Yuanxin Tian, Shiwei Zhang, Hongtao Lei, Yueming JiangAbstract:A heterologous competitive indirect enzyme-linked immunosorbent assay (ciELISA) for the determination of the furaltadone metabolite 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) was developed. AMOZ was derivatised with 2-(4-formylphenoxy) acetic acid or 2-(3-formylphenoxy) acetic acid to obtain two novel immunizing Haptens. The ability of these Haptens in producing specific polyclonal antibodies against the nitrophenyl derivative of AMOZ (NPAMOZ) was compared with that of traditional immunizing Haptens (derivatised AMOZ with 3-carboxybenzaldehyle or 4-carboxybenzaldehyle). The results indicated that the novel immunizing Haptens were able to produce antibodies with almost a two-fold improvement in sensitivity of the ciELISA for NPAMOZ in comparison with the existing antibody based ELISAs. The differences in sensitivity were explained by the molecular modeling of the lowest energy conformations of NPAMOZ and the Haptens. Another novel hapten, derivatised AMOZ with 2-oxoacetic acid, was synthesized and used as a heterologous coating hapten. The results showed that this strategy of using only a partial structure of the target molecule as the coating hapten was able to obtain a two to three-fold improvement in sensitivity. This study provided a modern approach for the development of an immunoassay with improved sensitivity for the metabolites of nitrofuran antibiotics.
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broad specificity immunoassay for o o diethyl organophosphorus pesticides application of molecular modeling to improve assay sensitivity and study antibody recognition
Analytical Chemistry, 2010Co-Authors: Yudong Shen, Hongtao Lei, Jinyi Yang, Hong Wang, Wenxu Zheng, Ross C Beier, Guimian Xie, Jiexian Dong, Zhigang She, Yuanming SunAbstract:A monoclonal antibody (mAb) against 4-(diethoxyphosphorothioyloxy)benzoic acid (hapten 1) was raised and used to develop a broad-specificity competitive indirect enzyme-linked immunosorbent assay (ciELISA) for 14 O,O-diethyl organophosphorus pesticides (OPs). Computer-assisted molecular modeling was used to model two-dimensional (2D) and three-dimensional (3D) quantitative structure−activity relationships (QSARs) to study antibody recognition. On the basis of insights obtained from the QSAR models, two heterologous coating Haptens, 4-(diethoxyphosphorothioylamino)butanoic acid (hapten 2) and 4-(diethoxyphosphorothioyloxy)-2-methylbenzoic acid (hapten 3) were designed, synthesized, and used to develop heterologous ciELISAs with significantly improved sensitivity. The heterologous ciELISA using hapten 2 as the coating hapten showed good sensitivity in a broad-specific manner for eight O,O-diethyl OPs and may be used as a screening method for the determination of these OPs. Our studies demonstrated that mole...
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hapten synthesis and antibody production for the development of a melamine immunoassay
Analytica Chimica Acta, 2010Co-Authors: Hongtao Lei, Yudong Shen, Yuanming Sun, Lijun Song, Jinyi Yang, Olivier P Chevallier, Simon A Haughey, Hong Wang, Christopher T ElliottAbstract:Abstract The incorporation of melamine into food products is banned but its misuse has been widely reported in both animal feeds and food. The development of a rapid screening immunoassay for monitoring of the substance is an urgent requirement. Two Haptens of melamine were synthesized by introducing spacer arms of different lengths and structures on the triazine ring of the analyte molecular structure. 6-Aminocaproic acid and 3-mercaptopropionic acid were reacted with 2-chloro-4,6-diamino-1,3,5-triazine (CAAT) to produce hapten 1 [3-(4,6-diamino-1,6-dihydro-1,3,5-triazin-2-ylamino) hexanoic acid] and hapten 2 [3-(4,6-diamino-1,6-dihydro-1,3,5-triazin-2-ylthio) propanoic acid], respectively. The molecular structures of the two Haptens were identified by 1 H nuclear magnetic resonance spectrometry, mass spectrometry and infrared spectrometry. An immunogen was prepared by coupling hapten 1 to bovine serum albumin (BSA). Two plate coating antigens were prepared by coupling both Haptens to egg ovalbumin (OVA). A competitive indirect enzyme-linked immunosorbent assay (ciELISA) was developed to evaluate homogeneous and heterogeneous assay formats. The results showed that polyclonal antibodies with high titers were obtained, and the heterogeneous immunoassay format demonstrated a better performance with an IC 50 of 70.6 ng mL −1 , a LOD of 2.6 ng mL −1 and a LOQ of 7.6 ng mL −1 . Except for cyromazine, no obvious cross-reactivity to common compounds was found. The data showed that the hapten synthesis was successful and the resultant antisera could be used in an immunoassay for the rapid and sensitive detection of this banned chemical.
Yueming Jiang - One of the best experts on this subject based on the ideXlab platform.
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novel hapten synthesis for antibody production and development of an enzyme linked immunosorbent assay for determination of furaltadone metabolite 3 amino 5 morpholinomethyl 2 oxazolidinone amoz
Talanta, 2013Co-Authors: Yudong Shen, Yuanming Sun, Katrina Campbell, Yuanxin Tian, Shiwei Zhang, Hongtao Lei, Yueming JiangAbstract:A heterologous competitive indirect enzyme-linked immunosorbent assay (ciELISA) for the determination of the furaltadone metabolite 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) was developed. AMOZ was derivatised with 2-(4-formylphenoxy) acetic acid or 2-(3-formylphenoxy) acetic acid to obtain two novel immunizing Haptens. The ability of these Haptens in producing specific polyclonal antibodies against the nitrophenyl derivative of AMOZ (NPAMOZ) was compared with that of traditional immunizing Haptens (derivatised AMOZ with 3-carboxybenzaldehyle or 4-carboxybenzaldehyle). The results indicated that the novel immunizing Haptens were able to produce antibodies with almost a two-fold improvement in sensitivity of the ciELISA for NPAMOZ in comparison with the existing antibody based ELISAs. The differences in sensitivity were explained by the molecular modeling of the lowest energy conformations of NPAMOZ and the Haptens. Another novel hapten, derivatised AMOZ with 2-oxoacetic acid, was synthesized and used as a heterologous coating hapten. The results showed that this strategy of using only a partial structure of the target molecule as the coating hapten was able to obtain a two to three-fold improvement in sensitivity. This study provided a modern approach for the development of an immunoassay with improved sensitivity for the metabolites of nitrofuran antibiotics.
