Preparation of monoclonal antibody against chloramphenicol and establishment of direct competitive ELISA for determination of chloramphenicol
Through immunization of BALB/c mice with synthesized CAP-ovalbumin (CAP-OVA), a hybridoma cell line, the 4C9 which could produce monoclonal antibody against chloramphenicol (CAP) and propagate stably, was established. With the myeloma cells, the ascites containing monclonal antibody (McAb) were prep...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Zhejiang University Press
2010-03-01
|
| Series: | 浙江大学学报. 农业与生命科学版 |
| Subjects: | |
| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2010.02.003 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849415819268718592 |
|---|---|
| author | CHEN Zheng-xian SHI Man-ling LI Yin-lai YE Xing-qian SHEN Li-rong |
| author_facet | CHEN Zheng-xian SHI Man-ling LI Yin-lai YE Xing-qian SHEN Li-rong |
| author_sort | CHEN Zheng-xian |
| collection | DOAJ |
| description | Through immunization of BALB/c mice with synthesized CAP-ovalbumin (CAP-OVA), a hybridoma cell line, the 4C9 which could produce monoclonal antibody against chloramphenicol (CAP) and propagate stably, was established. With the myeloma cells, the ascites containing monclonal antibody (McAb) were prepared. The indirect competitive ELISA (ciELISA) test revealed the heavy chain and light chain of McAb were IgG1 and κ, respectively. The ELISA titer of ascites was 1∶1 × 10<sup>6</sup>. The percentages of cross-activity to thiamphenicol, florfenicol and other antibiotics were all less than 0.01%. A horseradish peroxidase (HRP)-labeled antibody was synthesized by the sodium periodate reaction, then a direct competitive enzyme-linked immunosorbent assay (cdELISA) was developed. Quantization of the CAP was linear from 0.1 to 100 ng·mL<sup>-1</sup>, and the hemi-inhibitory concentration (IC<sub>50</sub>) was 5.81 ng·L<sup>-1</sup>. The recovery test showed that the detection limit for CAP was 0.1 ng·L<sup>-1</sup> in cdELISA. Comparative test showed that detected sensibility of the monoclonal antibody was nearly equivalent to the commercial CAP ELISA kit. |
| format | Article |
| id | doaj-art-ef37acf5f25e4026a2a47685709c5589 |
| institution | Kabale University |
| issn | 1008-9209 2097-5155 |
| language | English |
| publishDate | 2010-03-01 |
| publisher | Zhejiang University Press |
| record_format | Article |
| series | 浙江大学学报. 农业与生命科学版 |
| spelling | doaj-art-ef37acf5f25e4026a2a47685709c55892025-08-20T03:33:22ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552010-03-013613313810.3785/j.issn.1008-9209.2010.02.00310089209Preparation of monoclonal antibody against chloramphenicol and establishment of direct competitive ELISA for determination of chloramphenicolCHEN Zheng-xianSHI Man-lingLI Yin-laiYE Xing-qianSHEN Li-rongThrough immunization of BALB/c mice with synthesized CAP-ovalbumin (CAP-OVA), a hybridoma cell line, the 4C9 which could produce monoclonal antibody against chloramphenicol (CAP) and propagate stably, was established. With the myeloma cells, the ascites containing monclonal antibody (McAb) were prepared. The indirect competitive ELISA (ciELISA) test revealed the heavy chain and light chain of McAb were IgG1 and κ, respectively. The ELISA titer of ascites was 1∶1 × 10<sup>6</sup>. The percentages of cross-activity to thiamphenicol, florfenicol and other antibiotics were all less than 0.01%. A horseradish peroxidase (HRP)-labeled antibody was synthesized by the sodium periodate reaction, then a direct competitive enzyme-linked immunosorbent assay (cdELISA) was developed. Quantization of the CAP was linear from 0.1 to 100 ng·mL<sup>-1</sup>, and the hemi-inhibitory concentration (IC<sub>50</sub>) was 5.81 ng·L<sup>-1</sup>. The recovery test showed that the detection limit for CAP was 0.1 ng·L<sup>-1</sup> in cdELISA. Comparative test showed that detected sensibility of the monoclonal antibody was nearly equivalent to the commercial CAP ELISA kit.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2010.02.003chloramphenicolmonclonal antibodyhorseradish peroxidase-labeled antibodydirect competitive ELISA |
| spellingShingle | CHEN Zheng-xian SHI Man-ling LI Yin-lai YE Xing-qian SHEN Li-rong Preparation of monoclonal antibody against chloramphenicol and establishment of direct competitive ELISA for determination of chloramphenicol 浙江大学学报. 农业与生命科学版 chloramphenicol monclonal antibody horseradish peroxidase-labeled antibody direct competitive ELISA |
| title | Preparation of monoclonal antibody against chloramphenicol and establishment of direct competitive ELISA for determination of chloramphenicol |
| title_full | Preparation of monoclonal antibody against chloramphenicol and establishment of direct competitive ELISA for determination of chloramphenicol |
| title_fullStr | Preparation of monoclonal antibody against chloramphenicol and establishment of direct competitive ELISA for determination of chloramphenicol |
| title_full_unstemmed | Preparation of monoclonal antibody against chloramphenicol and establishment of direct competitive ELISA for determination of chloramphenicol |
| title_short | Preparation of monoclonal antibody against chloramphenicol and establishment of direct competitive ELISA for determination of chloramphenicol |
| title_sort | preparation of monoclonal antibody against chloramphenicol and establishment of direct competitive elisa for determination of chloramphenicol |
| topic | chloramphenicol monclonal antibody horseradish peroxidase-labeled antibody direct competitive ELISA |
| url | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2010.02.003 |
| work_keys_str_mv | AT chenzhengxian preparationofmonoclonalantibodyagainstchloramphenicolandestablishmentofdirectcompetitiveelisafordeterminationofchloramphenicol AT shimanling preparationofmonoclonalantibodyagainstchloramphenicolandestablishmentofdirectcompetitiveelisafordeterminationofchloramphenicol AT liyinlai preparationofmonoclonalantibodyagainstchloramphenicolandestablishmentofdirectcompetitiveelisafordeterminationofchloramphenicol AT yexingqian preparationofmonoclonalantibodyagainstchloramphenicolandestablishmentofdirectcompetitiveelisafordeterminationofchloramphenicol AT shenlirong preparationofmonoclonalantibodyagainstchloramphenicolandestablishmentofdirectcompetitiveelisafordeterminationofchloramphenicol |