学術雑誌論文

学術雑誌論文

text

Scientific Reports. 2017, 7(1), p. 5794

Efficient generation of single domain antibodies with high affinities and enhanced thermal stabilities

Nature Research

英語

text

2017-07-19

https://doi.org/10.1038/s41598-017-06277-x

Shinozaki, N., Hashimoto, R., Fukui, K. et al. Efficient generation of single domain antibodies with high affinities and enhanced thermal stabilities. Sci Rep 7, 5794 (2017). https://doi.org/10.1038/s41598-017-06277-x.

Single domain antibodies (sdAbs), made of natural single variable regions of camelid or cartilaginous fish antibodies, or unpaired variable regions of mouse or human IgGs, are some of the more promising biologic modalities. However, such conventional sdAbs have difficulties of either using unwieldy animals for immunization or having high affinity deficiencies. Herein, we offer a versatile method to generate rabbit variable domain of heavy chain (rVH) derived sdAbs with high affinities (K D values of single digit nM or less) and enhanced thermal stabilities (equal to or even higher than those of camelid derived sdAbs). It was found that a variety of rVH binders, including those with high affinities, were efficiently acquired using an rVH-displaying phage library produced at a low temperature of 16 °C. By a simple method to introduce an additional disulfide bond, their unfolding temperatures were increased by more than 20 °C without severe loss of binding affinity. Differential scanning calorimetry analysis suggested that this highly efficient thermal stabilization was mainly attributed to the entropic contribution and unique thermodynamic character of the rVHs.

© 2017 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License.

79025

Efficient generation of single domain antibodies with high affinities and enhanced thermal stabilities

Nature Research

英語

2017-07-19

Scientific Reports

7

1

5794

https://doi.org/10.1038/s41598-017-06277-x

28725057

https://hdl.handle.net/11094/79025