Neutralising Effects of Different Antibodies on Clostridioides difficile Toxins TcdA and TcdB in a Translational Approach
Abstract
:1. Introduction
2. Results
2.1. Recombinant and Natural sIgA Antibodies Were Successfully Purified
2.2. Recombinant and Natural Antibodies Mainly Bind to TcdB
2.3. Recombinant Antibodies Exhibited Superior TcdB Neutralisation Potential Compared to Natural sIgA Molecules
2.4. Successful Establishment of Apical-Out and Floating Basal-Out Organoids
2.5. Considerably Different Effects of TcdA and TcdB on Basal-Out and Apical-Out Organoids
3. Discussion
4. Materials and Methods
4.1. Recombinant Monoclonal sIgA
4.2. Isolation of Caprine Whey sIgA
4.3. ELISA
4.4. Organoid Culture
4.5. Generation of Organoid-Derived Monolayers
4.6. Sulforhodamine B Cytotoxicity Assay
4.7. Generation of Floating Basal-Out and Apical-Out Organoids
4.8. DAPI/Phalloidin Staining
4.9. FITC-Dextran Barrier Integrity Assay
4.10. Statistics
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Csukovich, G.; Kramer, N.; Pratscher, B.; Gotic, I.; Freund, P.; Hahn, R.; Himmler, G.; Brandt, S.; Burgener, I.A. Neutralising Effects of Different Antibodies on Clostridioides difficile Toxins TcdA and TcdB in a Translational Approach. Int. J. Mol. Sci. 2023, 24, 3867. https://doi.org/10.3390/ijms24043867
Csukovich G, Kramer N, Pratscher B, Gotic I, Freund P, Hahn R, Himmler G, Brandt S, Burgener IA. Neutralising Effects of Different Antibodies on Clostridioides difficile Toxins TcdA and TcdB in a Translational Approach. International Journal of Molecular Sciences. 2023; 24(4):3867. https://doi.org/10.3390/ijms24043867
Chicago/Turabian StyleCsukovich, Georg, Nina Kramer, Barbara Pratscher, Ivana Gotic, Patricia Freund, Rainer Hahn, Gottfried Himmler, Sabine Brandt, and Iwan Anton Burgener. 2023. "Neutralising Effects of Different Antibodies on Clostridioides difficile Toxins TcdA and TcdB in a Translational Approach" International Journal of Molecular Sciences 24, no. 4: 3867. https://doi.org/10.3390/ijms24043867