Screening of Neutralizing Antibodies against FaeG Protein of Enterotoxigenic Escherichia coli
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Median Lethal Dose and Minimum Lethal Dose Test of Isolated ETEC Strains
2.2. Immunization of Mice
2.3. Expression and Fluorescent Labeling of F4ac FaeG Protein in ETEC
2.4. Indirect Enzyme-Linked Immunosorbent Assay (iELISA)
2.5. Fluorescence-Activated Cell Sorting
2.6. Amplification of Variable Region and Construction of Expression Plasmids
2.7. Expression and Analyses of Anti-rFaeG IgG mAbs
2.8. Adhesion Assay of ETEC
2.9. Anti-rFaeG IgG mAb Neutralization Assay in Mice
3. Results
3.1. Soluble Expression and Immune Response of rFaeG Protein in Mice
3.2. Sorting of rFaeG-Specific Mouse Memory B Cells
3.3. Preparation of Anti-rFaeG IgG mAbs
3.4. Neutralizing Activity of Anti-rFaeG IgG mAbs In Vitro
3.5. Neutralizing Activity of Anti-rFaeG IgG mAbs In Vivo
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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Primers | Sequence (5′-3′) a |
---|---|
VH1-F | (CCGATCCAGCCTCCGGAC)MAGCTTCAGGAGTCRGGACC |
VH2-F | (CCGATCCAGCCTCCGGAC)CAGCTGAAGSASTCAGGACC |
VH3-F | (CCGATCCAGCCTCCGGAC)MWGSKGGTGGAGTCTGGGGGA |
VH4-F | (CCGATCCAGCCTCCGGAC)ARSSTGGWGGAATCTGGAGGA |
VH5-F | (CCGATCCAGCCTCCGGAC)ARGSTGRTSGAGTCTGGAGG |
VH6-F | (CCGATCCAGCCTCCGGAC)CARSYGCAGCARYCTGGG |
VH7-F | (CCGATCCAGCCTCCGGAC)CAGYTGSWGCARTCTGGA |
VH8-F | (CCGATCCAGCCTCCGGAC)CAGCTGCAGCAGTCWGTG |
VH9-F | (CCGATCCAGCCTCCGGAC)MASYTGSWGGWGWCTGGAGG |
VH10-F | (CCGATCCAGCCTCCGGAC)CAGMTSCAGCAGYCTGG |
1st reverse γ | CKYGGTSYTGCTGGCYGGGTG |
2nd reverse γ | GCTCAGGGAARTAGCCCTTGAC |
Vκ1-F | (CCGATCCAGCCTCCGGAC)CAGACRTCMAGATRAYCCAGWCTMCA |
Vκ2-F | (CCGATCCAGCCTCCGGAC)CARAMATTKTGCTGACYCARTYTCC |
Vκ3-F | (CCGATCCAGCCTCCGGAC)CAGATRYTKTGATGACCCAAACTCCA |
Vκ4-F | (CCGATCCAGCCTCCGGAC)CASRAAWTSTTCTCWYMCAGTGTCC |
1st reverse κ | CTAACACTCATTCCTGTTGAAGC |
2nd reverse κ | TGGGAAGATGGATACAGTT |
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Tian, Y.; Lu, S.; Zhou, S.; Li, Z.; Guan, S.; Chen, H.; Song, Y. Screening of Neutralizing Antibodies against FaeG Protein of Enterotoxigenic Escherichia coli. Vet. Sci. 2024, 11, 419. https://doi.org/10.3390/vetsci11090419
Tian Y, Lu S, Zhou S, Li Z, Guan S, Chen H, Song Y. Screening of Neutralizing Antibodies against FaeG Protein of Enterotoxigenic Escherichia coli. Veterinary Sciences. 2024; 11(9):419. https://doi.org/10.3390/vetsci11090419
Chicago/Turabian StyleTian, Yang, Sijia Lu, Saisai Zhou, Zhen Li, Shuaiyin Guan, Huanchun Chen, and Yunfeng Song. 2024. "Screening of Neutralizing Antibodies against FaeG Protein of Enterotoxigenic Escherichia coli" Veterinary Sciences 11, no. 9: 419. https://doi.org/10.3390/vetsci11090419