Isolation of Anti-Ricin Protective Antibodies Exhibiting High Affinity from Immunized Non-Human Primates
Abstract
:1. Introduction
2. Results
2.1. Immunization and Characterization of Elicited Antibodies
2.2. Isolation of Anti-Ricin Antibodies
2.3. Epitope Binning
2.4. In Vitro Neutralization of Ricin Cytotoxicity
2.5. Affinity of the Anti-Ricin Antibodies
2.6. Post-Exposure Treatment of Ricin Intoxication
3. Discussion
4. Experimental Section
4.1. Animal Immunization
4.2. Primers Design
4.3. scFv Library Construction
4.4. Library Screening
4.5. ELISA
4.6. Fingerprint Analysis
4.7. Nucleic Acid Analysis
4.8. Producing Full-Length Antibodies
4.9. Affinity Measurements and Epitope Binning
4.10. In Vitro Ricin Neutralization Assay
4.11. In Vivo Protection Assay
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Antibody | Vaccine a | Binding b | ED50 (ng/mL) c | kon (1/Ms) | koff (1/s) | KD (nM) |
---|---|---|---|---|---|---|
MH1 | subunit | RTA | 200 d | 9 × 104 | <1 × 10−7 | <0.001 |
MH2 | subunit | RTB | 5200 | 18 × 104 | 1 × 10−5 | 0.6 |
MH36 | holotoxin | RTA | 27,000 | 11 × 104 | <1 × 10−7 | <0.001 |
MH49 | holotoxin | RTA | 1200 | 7 × 104 | <1 × 10−7 | <0.001 |
MH67 | holotoxin | RTA | 52,000 | 9 × 104 | 8 × 10−4 | 9.0 |
MH73 | subunit | RTB | 11,000 | 11 × 104 | <1 × 10−7 | <0.001 |
MH74 | subunit | RTA | 83,000 d | 15 × 104 | 7 × 10−3 | 45 |
MH75 | subunit | RTB | 150 d | 35 × 104 | <1 × 10−7 | <0.001 |
MH76 | subunit | RTB | 10,500 | 11 × 104 | 3 × 10−4 | 3.0 |
MH77 | subunit | RTB | 500 | 17 × 104 | 1 × 10−7 | 0.001 |
Antibody | VH | VL | |||
---|---|---|---|---|---|
V | D | J | V | J | |
MH1 a | IGHV4.40 | IGHD2-4*01 | IGHJ1*01 | IGLV3.46 c | IGLJ3 |
MH2 a | IGHV3.15 | IGHD5-3*01 | IGHJ5-1*01 | IGLV3.46 c | IGLJ2 |
MH36 b | IGHV3.9 | IGHD4-2*01 | IGHJ4*01 | IGLV1.25 c | IGLJ2 |
MH49 b | IGHV4.11 | IGHD4-2*01 | IGHJ3*01 | IGLV1.27 c | IGLJx1 |
MH67 b | IGHV4.11 | IGHD4-2*01 | IGHJ3*01 | IGKV1.9 d | IGKJ2 |
MH73 a | IGHV4.11 | IGHD1-2*01 | IGHJ5-1*01 | IGKV1.52 d | IGKJ2 |
MH74 a | IGHV5.7 | IGHD3-3*01 | IGHJ5-2*02 | IGLV1.30 c | IGLJ3 |
MH75 a | IGHV3.15 | IGHD5-3*01 | IGHJ5-1*01 | IGLV3.46 c | IGLJ2 |
MH76 a | IGHV3.15 | IGHD3-1*01 | IGHJ4*01 | IGLV3.46 c | IGLJ3 |
MH77 a | IGHV4.40 | IGHD4-4*01 | IGHJ5-1*01 | IGKV1.7 d | IGKJ2 |
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Noy-Porat, T.; Rosenfeld, R.; Ariel, N.; Epstein, E.; Alcalay, R.; Zvi, A.; Kronman, C.; Ordentlich, A.; Mazor, O. Isolation of Anti-Ricin Protective Antibodies Exhibiting High Affinity from Immunized Non-Human Primates. Toxins 2016, 8, 64. https://doi.org/10.3390/toxins8030064
Noy-Porat T, Rosenfeld R, Ariel N, Epstein E, Alcalay R, Zvi A, Kronman C, Ordentlich A, Mazor O. Isolation of Anti-Ricin Protective Antibodies Exhibiting High Affinity from Immunized Non-Human Primates. Toxins. 2016; 8(3):64. https://doi.org/10.3390/toxins8030064
Chicago/Turabian StyleNoy-Porat, Tal, Ronit Rosenfeld, Naomi Ariel, Eyal Epstein, Ron Alcalay, Anat Zvi, Chanoch Kronman, Arie Ordentlich, and Ohad Mazor. 2016. "Isolation of Anti-Ricin Protective Antibodies Exhibiting High Affinity from Immunized Non-Human Primates" Toxins 8, no. 3: 64. https://doi.org/10.3390/toxins8030064
APA StyleNoy-Porat, T., Rosenfeld, R., Ariel, N., Epstein, E., Alcalay, R., Zvi, A., Kronman, C., Ordentlich, A., & Mazor, O. (2016). Isolation of Anti-Ricin Protective Antibodies Exhibiting High Affinity from Immunized Non-Human Primates. Toxins, 8(3), 64. https://doi.org/10.3390/toxins8030064