期刊
NANO LETTERS
卷 19, 期 8, 页码 5524-5529出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.nanolett.9b02062
关键词
Antibodies; biophysics; disulfide bond; nanobody; single-molecule force spectroscopy
类别
资金
- University of Basel
- ERC [MMA-715207]
- NCCR in Molecular Systems Engineering
- SNSF [200021_175478]
- ETH Zurich
- Swiss National Science Foundation (SNF) [200021_175478] Funding Source: Swiss National Science Foundation (SNF)
Single-domain VHH antibodies are promising reagents for medical therapy. A conserved disulfide bond within the VHH framework region is known to be critical for thermal stability, however, no prior studies have investigated its influence on the stability of VHH antibody-antigen complexes under mechanical load. Here, we used single-molecule force spectroscopy to test the influence of a VHH domain's conserved disulfide bond on the mechanical strength of the interaction with its antigen mCherry. We found that although removal of the disulfide bond through cysteine-to-alanine mutagenesis significantly lowered VHH domain denaturation temperature, it had no significant impact on the mechanical strength of the VHH:mCherry interaction with complex rupture occurring at similar to 60 pN at 10(3)-10(4) pN/sec regardless of disulfide bond state. These results demonstrate that mechanostable binding interactions can be built on molecular scaffolds that may be thermodynamically compromised at equilibrium.
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