Production of polyclonal antibody against the recombinant PirBvp protein of Vibrio parahaemolyticus.
| Title: | Production of polyclonal antibody against the recombinant PirBvp protein of Vibrio parahaemolyticus. |
|---|---|
| Authors: | Duong ND; University of Science, 227 Nguyen Van Cu Street, Ward 4, District 5, Ho Chi Minh City, Vietnam.; Vietnam National University, Ho Chi Minh City, Vietnam.; Pasteur Institute, 167 Pasteur Street, Vo Thi Sau Ward, District 3, Ho Chi Minh City, Vietnam.; Nguyen-Phuoc KH; University of Science, 227 Nguyen Van Cu Street, Ward 4, District 5, Ho Chi Minh City, Vietnam.; Vietnam National University, Ho Chi Minh City, Vietnam.; Do KT; Pasteur Institute, 167 Pasteur Street, Vo Thi Sau Ward, District 3, Ho Chi Minh City, Vietnam.; Nguyen NT; Pasteur Institute, 167 Pasteur Street, Vo Thi Sau Ward, District 3, Ho Chi Minh City, Vietnam.; Tran TL; University of Science, 227 Nguyen Van Cu Street, Ward 4, District 5, Ho Chi Minh City, Vietnam.; Vietnam National University, Ho Chi Minh City, Vietnam.; Tran-Van H; University of Science, 227 Nguyen Van Cu Street, Ward 4, District 5, Ho Chi Minh City, Vietnam. tvhieu@hcmus.edu.vn.; Vietnam National University, Ho Chi Minh City, Vietnam. tvhieu@hcmus.edu.vn. |
| Source: | Journal, genetic engineering & biotechnology [J Genet Eng Biotechnol] 2021 May 11; Vol. 19 (1), pp. 70. Date of Electronic Publication: 2021 May 11. |
| Publication Type: | Journal Article |
| Language: | English |
| Journal Info: | Publisher: Elsevier B.V Country of Publication: Netherlands NLM ID: 101317150 Publication Model: Electronic Cited Medium: Internet ISSN: 2090-5920 (Electronic) Linking ISSN: 1687157X NLM ISO Abbreviation: J Genet Eng Biotechnol Subsets: PubMed not MEDLINE |
| Imprint Name(s): | Publication: 2024- : [Amsterdam, Netherlands] : Elsevier B.V.; Original Publication: [Cairo] : Academy of Scientific Research and Technology, Information and Scientific Services Sector, National Information and Documentation Center |
| Abstract: | Background: Acute hepatopancreatic necrosis disease (AHPND) is caused by toxin-producing strains of Vibrio parahaemolyticus which contain deadly binary toxins PirAvp and PirBvp encoded in pVA1 plasmid. The polyclonal antibodies against PirBvp protein could be used to develop immunochromatographic test strip for in-field diagnosis of AHPND.; Results: In this study, PirBvp gene was amplified, cloned, and expressed in E. coli. The expressed protein was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot probed with 6xHis antibodies. Then, the recombinant PirBvp (rPirBvp) was purified using Ni-Sepharose column. Rabbits were immunized with the purified rPirBvp, and produced antibodies were analyzed using Ouchterlony double immunodiffusion. The antibody titration and antibody purification were performed by ELISA and affinity chromatography, respectively. Finally, antibody specificity and sensitivity were evaluated by dot blotting. The present study showed a high titer of polyclonal antibodies in rabbit serum after immunization and the titer increased steadily during the immunization schedule. The highest titer of antibody reached up to 2,560,000 with LOD of 0.1 ng/mL. The purified antibodies showed no cross-reactivity with proteins from other Vibrio species, and the detection threshold ranged from 6.25 to 12.5 ng toxin/dot.; Conclusion: This study highlights the production of high titer and specific polyclonal antibodies as an initial material towards the development of lateral-flow strip test. |
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| Grant Information: | 19/2018/HĐ-KHCN-TNB.ĐT/14-19/C31 The Mekong Delta Program Office |
| Contributed Indexing: | Keywords: AHPND; ELISA; PirBvp protein; Polyclonal antibodies; Recombinant protein; Vibrio parahaemolyticus |
| Entry Date(s): | Date Created: 20210512 Latest Revision: 20240402 |
| Update Code: | 20260130 |
| PubMed Central ID: | PMC8113428 |
| DOI: | 10.1186/s43141-021-00172-9 |
| PMID: | 33977321 |
| Database: | MEDLINE |
Journal Article