JIANG JinQing; ZHANG HaiTang; YAO SiXin; WANG ZiLiang; WANG JianHua

doi:10.3967/0895-3988.2011.02.012

Production and Characterisation of Monoclonal Antibodies against 19-Nortestosterone

JIANG JinQing, ZHANG HaiTang, YAO SiXin, WANG ZiLiang, WANG JianHua

文章导航 > Biomedical and Environmental Sciences > 2011 > 24(2): 172-179

JIANG JinQing, ZHANG HaiTang, YAO SiXin, WANG ZiLiang, WANG JianHua. Production and Characterisation of Monoclonal Antibodies against 19-Nortestosterone[J]. Biomedical and Environmental Sciences, 2011, 24(2): 172-179. doi: 10.3967/0895-3988.2011.02.012

Citation:

JIANG JinQing, ZHANG HaiTang, YAO SiXin, WANG ZiLiang, WANG JianHua. Production and Characterisation of Monoclonal Antibodies against 19-Nortestosterone[J]. Biomedical and Environmental Sciences, 2011, 24(2): 172-179. doi: 10.3967/0895-3988.2011.02.012

doi: 10.3967/0895-3988.2011.02.012

基金项目: the Henan Innovation Project for University Prominent Research Talents(2010HASTIT026)

Production and Characterisation of Monoclonal Antibodies against 19-Nortestosterone

College of Veterinary Medicine, Northwest A&F University, Yangling 712100, Shanxi, China;College of Animal Science, Henan Institute of Science and Technology, Xinxiang 453003, Henan, China
College of Animal Science, Henan Institute of Science and Technology, Xinxiang 453003, Henan, China
College of Veterinary Medicine, Northwest A&F University, Yangling 712100, Shanxi, China

Funds: the Henan Innovation Project for University Prominent Research Talents(2010HASTIT026)

摘要: Objective To produce anti-19-Nortestosterone (NT) monoclonal antibodies and identify their immunological characteristics. Methods Hybridomas were prepared by fusing NSO mouse myeloma cells with splenocytes isolated from immunized BALB/c mice. Noncompetitive and competitive indirect ELISA were employed to screen positive cell clones. A caprylic acid ammonium sulphate (CAAP) method was used to purify NT mAb, and the Batty saturation method was used to determine the affinity constant (Kaff). Results Five hybridorna cell lines, named NT-1, NT-2, NT-3, NT-4, and NT-5, were identified and their corresponding mAbs were of the IgG<,1> isotype with a k light chain. The Kaffs of all mAbs were between 2.6 and 4.7×10<'9> L/mol. The titers and IC<,50> values of purified ascite fluids were in the range of (0.64-2.56)×10<'5> and (0.55-1.0) ng/mL, respectively. Of all the cross-reacting steroids, α-NT was the most reactive with the mAbs at 62% with NT-1 mAb and 64% with NT-2 mAb. Negligible cross-reactivity (<0.01%) with other steroids was observed. Conclusion The establishment of these hybridomas allows the potential development of a rapid test kit, and may provide an alternative method for the detection of NT residues in food producing animals.
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Abstract: Objective To produce anti-19-Nortestosterone (NT) monoclonal antibodies and identify their immunological characteristics. Methods Hybridomas were prepared by fusing NSO mouse myeloma cells with splenocytes isolated from immunized BALB/c mice. Noncompetitive and competitive indirect ELISA were employed to screen positive cell clones. A caprylic acid ammonium sulphate (CAAP) method was used to purify NT mAb, and the Batty saturation method was used to determine the affinity constant (Kaff). Results Five hybridorna cell lines, named NT-1, NT-2, NT-3, NT-4, and NT-5, were identified and their corresponding mAbs were of the IgG<,1> isotype with a k light chain. The Kaffs of all mAbs were between 2.6 and 4.7×10<'9> L/mol. The titers and IC<,50> values of purified ascite fluids were in the range of (0.64-2.56)×10<'5> and (0.55-1.0) ng/mL, respectively. Of all the cross-reacting steroids, α-NT was the most reactive with the mAbs at 62% with NT-1 mAb and 64% with NT-2 mAb. Negligible cross-reactivity (<0.01%) with other steroids was observed. Conclusion The establishment of these hybridomas allows the potential development of a rapid test kit, and may provide an alternative method for the detection of NT residues in food producing animals.
- 19-Nortestosterone /
- Hybridoma cell lines /
- Monoclonal antibody /
- Characterization /
- Identification

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Production and Characterisation of Monoclonal Antibodies against 19-Nortestosterone

doi: 10.3967/0895-3988.2011.02.012

基金项目: the Henan Innovation Project for University Prominent Research Talents(2010HASTIT026)

刊出日期: 2011-04-20

关键词:

19-Nortestosterone /

Hybridoma cell lines /

Monoclonal antibody /

Characterization /

Identification

摘要: Objective To produce anti-19-Nortestosterone (NT) monoclonal antibodies and identify their immunological characteristics. Methods Hybridomas were prepared by fusing NSO mouse myeloma cells with splenocytes isolated from immunized BALB/c mice. Noncompetitive and competitive indirect ELISA were employed to screen positive cell clones. A caprylic acid ammonium sulphate (CAAP) method was used to purify NT mAb, and the Batty saturation method was used to determine the affinity constant (Kaff). Results Five hybridorna cell lines, named NT-1, NT-2, NT-3, NT-4, and NT-5, were identified and their corresponding mAbs were of the IgG<,1> isotype with a k light chain. The Kaffs of all mAbs were between 2.6 and 4.7×10<'9> L/mol. The titers and IC<,50> values of purified ascite fluids were in the range of (0.64-2.56)×10<'5> and (0.55-1.0) ng/mL, respectively. Of all the cross-reacting steroids, α-NT was the most reactive with the mAbs at 62% with NT-1 mAb and 64% with NT-2 mAb. Negligible cross-reactivity (<0.01%) with other steroids was observed. Conclusion The establishment of these hybridomas allows the potential development of a rapid test kit, and may provide an alternative method for the detection of NT residues in food producing animals.

English Abstract

Production and Characterisation of Monoclonal Antibodies against 19-Nortestosterone

College of Veterinary Medicine, Northwest A&F University, Yangling 712100, Shanxi, China;College of Animal Science, Henan Institute of Science and Technology, Xinxiang 453003, Henan, China

College of Animal Science, Henan Institute of Science and Technology, Xinxiang 453003, Henan, China

College of Veterinary Medicine, Northwest A&F University, Yangling 712100, Shanxi, China

Funds: the Henan Innovation Project for University Prominent Research Talents(2010HASTIT026)

Publish Date: 2011-04-20

Keywords:

Abstract: Objective To produce anti-19-Nortestosterone (NT) monoclonal antibodies and identify their immunological characteristics. Methods Hybridomas were prepared by fusing NSO mouse myeloma cells with splenocytes isolated from immunized BALB/c mice. Noncompetitive and competitive indirect ELISA were employed to screen positive cell clones. A caprylic acid ammonium sulphate (CAAP) method was used to purify NT mAb, and the Batty saturation method was used to determine the affinity constant (Kaff). Results Five hybridorna cell lines, named NT-1, NT-2, NT-3, NT-4, and NT-5, were identified and their corresponding mAbs were of the IgG<,1> isotype with a k light chain. The Kaffs of all mAbs were between 2.6 and 4.7×10<'9> L/mol. The titers and IC<,50> values of purified ascite fluids were in the range of (0.64-2.56)×10<'5> and (0.55-1.0) ng/mL, respectively. Of all the cross-reacting steroids, α-NT was the most reactive with the mAbs at 62% with NT-1 mAb and 64% with NT-2 mAb. Negligible cross-reactivity (<0.01%) with other steroids was observed. Conclusion The establishment of these hybridomas allows the potential development of a rapid test kit, and may provide an alternative method for the detection of NT residues in food producing animals.

Citation: