Preparation and Application of Clostridium perfringens Alpha Toxin Nanobodies
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Preparation of Recombinant CPA Protein
2.2. Screening and Preparation of CPA Nanobodies
2.3. Evaluation of CPA-VHH Characteristics
2.4. Neutralization of CPA-VHH on Phospholipase C and Erythrocyte Hemolyticity
2.5. Determination of the Half-Maximal Inhibitory Concentration (IC50) of CPA in Vero Cells
2.6. Investigation of CPA-VHH’s Neutralizing Effect on CPA in Vero Cells
2.7. Determination of Median Lethal Dose (LD50) for CPA
2.8. Biological Effects of CPA-VHH in Mice Exposed to CPA
2.9. Total RNA Isolation and Quantitative Real-Time PCR Analysis
2.10. Immunohistochemistry (IHC)
2.11. Statistical Analysis
3. Results
3.1. Preparation of CPA Recombinant Protein and CPA-VHH
3.2. Evaluation of CPA-VHH Characteristics
3.3. Neutralizing Effect of CPA-VHH on Phospholipase C and Erythrocyte Hemolyticity
3.4. Neutralizing Effect of CPA-VHH on CPA in Vero Cells
3.5. Effects of CPA-VHH in Terms of Neutralization of CPA and Prevention and Treatment of Infection with CPA in Mice
3.6. Protective Effect of CPA-VHH on Intestinal Mucosa during CPA Infection
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Primer Name | Primer Sequences (5′-3′) |
---|---|
ZO-1-F | GCCGCTAAGAGCACAGCAA |
ZO-1-R | GCCCTCCTTTTAACACATCAGA |
Occludin-F | TTGAAAGTCCACCTCCTTACAGA |
Occludin-R | CCGGATAAAAAGAGTACGCTGG |
Claudin-1-F | GCCTTGATGGTAATTGGCATCC |
Claudin-1-R | GGCCACTAATGTCGCCAGAC |
IL-1β-F | TTCAGGCAGGCAGTATCACTC |
IL-1β-R | GAAGGTCCACGGGAAAGACAC |
IL-6-F | TCCAGTTGCCTTCTTGGGAC |
IL-6-R | GACAGGTCTGTTGGGAGTGG |
IL-7-F | TTCCTCCACTGATCCTTGTTCT |
IL-7-R | AGCAGCTTCCTTTGTATCATCAC |
IL-8-F | ATGCCCTCTATTCTGCCAGAT |
IL-8-R | GTGCTCCGGTTGTATAAGATGAC |
TNF-α-F | CCCTCACACTCAGATCATCTTCT |
TNF-α-R | GCTACGACGTGGGCTACAG |
INF-γ-F | ATGAACGCTACACACTGCATC |
INF-γ-R | CCATCCTTTTGCCAGTTCCTC |
β-actin-F | TTGCTGACAGGATGCAGAAG |
β-actin-R | ACATCTGCTGGAAGGTGGAC |
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Jia, Q.; Ren, H.; Zhang, S.; Yang, H.; Gao, S.; Fan, R. Preparation and Application of Clostridium perfringens Alpha Toxin Nanobodies. Vet. Sci. 2024, 11, 381. https://doi.org/10.3390/vetsci11080381
Jia Q, Ren H, Zhang S, Yang H, Gao S, Fan R. Preparation and Application of Clostridium perfringens Alpha Toxin Nanobodies. Veterinary Sciences. 2024; 11(8):381. https://doi.org/10.3390/vetsci11080381
Chicago/Turabian StyleJia, Qiong, Hongrui Ren, Shuyin Zhang, Haoyu Yang, Shuaipeng Gao, and Ruiwen Fan. 2024. "Preparation and Application of Clostridium perfringens Alpha Toxin Nanobodies" Veterinary Sciences 11, no. 8: 381. https://doi.org/10.3390/vetsci11080381