Delicate balance among thermal stability, binding affinity, and conformational space explored by single-domain VHH antibodies
Abstract The high binding affinities and specificities of antibodies have led to their use as drugs and biosensors. Single-domain VHH antibodies exhibit high specificity and affinity but have higher stability and solubility than conventional antibodies as they are single-domain proteins. In this wor...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2021-10-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-021-98977-8 |
| _version_ | 1818575119667167232 |
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| author | Emina Ikeuchi Daisuke Kuroda Makoto Nakakido Akikazu Murakami Kouhei Tsumoto |
| author_facet | Emina Ikeuchi Daisuke Kuroda Makoto Nakakido Akikazu Murakami Kouhei Tsumoto |
| author_sort | Emina Ikeuchi |
| collection | DOAJ |
| description | Abstract The high binding affinities and specificities of antibodies have led to their use as drugs and biosensors. Single-domain VHH antibodies exhibit high specificity and affinity but have higher stability and solubility than conventional antibodies as they are single-domain proteins. In this work, based on physicochemical measurements and molecular dynamics (MD) simulations, we have gained insight that will facilitate rational design of single-chain VHH antibodies. We first assessed two homologous VHH antibodies by differential scanning calorimetry (DSC); one had a high (64.8 °C) and the other a low (58.6 °C) melting temperature. We then generated a series of the variants of the low stability antibody and analyzed their thermal stabilities by DSC and characterized their structures through MD simulations. We found that a single mutation that resulted in 8.2 °C improvement in melting temperature resulted in binding affinity an order of magnitude lower than the parent antibody, likely due to a shift of conformational space explored by the single-chain VHH antibody. These results suggest that the delicate balance among conformational stability, binding capability, and conformational space explored by antibodies must be considered in design of fully functional single-chain VHH antibodies. |
| first_indexed | 2024-12-15T00:36:01Z |
| format | Article |
| id | doaj.art-94c6e8e43a92458194b1a6a09e1f51ed |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-15T00:36:01Z |
| publishDate | 2021-10-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-94c6e8e43a92458194b1a6a09e1f51ed2022-12-21T22:41:48ZengNature PortfolioScientific Reports2045-23222021-10-011111910.1038/s41598-021-98977-8Delicate balance among thermal stability, binding affinity, and conformational space explored by single-domain VHH antibodiesEmina Ikeuchi0Daisuke Kuroda1Makoto Nakakido2Akikazu Murakami3Kouhei Tsumoto4Department of Bioengineering, School of Engineering, The University of TokyoDepartment of Bioengineering, School of Engineering, The University of TokyoDepartment of Bioengineering, School of Engineering, The University of TokyoDepartment of Parasitology and Immunopathoetiology, Graduate School of Medicine, University of the RyukyusDepartment of Bioengineering, School of Engineering, The University of TokyoAbstract The high binding affinities and specificities of antibodies have led to their use as drugs and biosensors. Single-domain VHH antibodies exhibit high specificity and affinity but have higher stability and solubility than conventional antibodies as they are single-domain proteins. In this work, based on physicochemical measurements and molecular dynamics (MD) simulations, we have gained insight that will facilitate rational design of single-chain VHH antibodies. We first assessed two homologous VHH antibodies by differential scanning calorimetry (DSC); one had a high (64.8 °C) and the other a low (58.6 °C) melting temperature. We then generated a series of the variants of the low stability antibody and analyzed their thermal stabilities by DSC and characterized their structures through MD simulations. We found that a single mutation that resulted in 8.2 °C improvement in melting temperature resulted in binding affinity an order of magnitude lower than the parent antibody, likely due to a shift of conformational space explored by the single-chain VHH antibody. These results suggest that the delicate balance among conformational stability, binding capability, and conformational space explored by antibodies must be considered in design of fully functional single-chain VHH antibodies.https://doi.org/10.1038/s41598-021-98977-8 |
| spellingShingle | Emina Ikeuchi Daisuke Kuroda Makoto Nakakido Akikazu Murakami Kouhei Tsumoto Delicate balance among thermal stability, binding affinity, and conformational space explored by single-domain VHH antibodies Scientific Reports |
| title | Delicate balance among thermal stability, binding affinity, and conformational space explored by single-domain VHH antibodies |
| title_full | Delicate balance among thermal stability, binding affinity, and conformational space explored by single-domain VHH antibodies |
| title_fullStr | Delicate balance among thermal stability, binding affinity, and conformational space explored by single-domain VHH antibodies |
| title_full_unstemmed | Delicate balance among thermal stability, binding affinity, and conformational space explored by single-domain VHH antibodies |
| title_short | Delicate balance among thermal stability, binding affinity, and conformational space explored by single-domain VHH antibodies |
| title_sort | delicate balance among thermal stability binding affinity and conformational space explored by single domain vhh antibodies |
| url | https://doi.org/10.1038/s41598-021-98977-8 |
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