English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Exploiting sequence and stability information for directing nanobody stability engineering

Kunz, P., Flock, T., Soler, N., Zaiss, M., Vincke, C., Sterckx, Y., et al. (2017). Exploiting sequence and stability information for directing nanobody stability engineering. Biochimica et Biophysica Acta: General Subjects, 1861(9), 2196-2205. doi:10.1016/j.bbagen.2017.06.014.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/21.11116/0000-0000-C2B4-9 Version Permalink: http://hdl.handle.net/21.11116/0000-0000-C2B5-8
Genre: Journal Article

Files

show Files

Locators

show
hide
Locator:
Link (Any fulltext)
Description:
-

Creators

show
hide
 Creators:
Kunz, P, Author
Flock, T, Author
Soler, N, Author
Zaiss, M1, 2, Author              
Vincke, C, Author
Sterckx, Y, Author
Kastelic, D, Author
Muyldermans, S, Author
Hoheisel, JE, Author
Affiliations:
1Max Planck Institute for Biological Cybernetics, Max Planck Society, ou_1497794              
2Department High-Field Magnetic Resonance, Max Planck Institute for Biological Cybernetics, Max Planck Society, ou_1497796              

Content

show
hide
Free keywords: -
 Abstract: Background Variable domains of camelid heavy-chain antibodies, commonly named nanobodies, have high biotechnological potential. In view of their broad range of applications in research, diagnostics and therapy, engineering their stability is of particular interest. One important aspect is the improvement of thermostability, because it can have immediate effects on conformational stability, protease resistance and aggregation propensity of the protein. Methods We analyzed the sequences and thermostabilities of 78 purified nanobody binders. From this data, potentially stabilizing amino acid variations were identified and studied experimentally. Results Some mutations improved the stability of nanobodies by up to 6.1 °C, with an average of 2.3 °C across eight modified nanobodies. The stabilizing mechanism involves an improvement of both conformational stability and aggregation behavior, explaining the variable degree of stabilization in individual molecules. In some instances, variations predicted to be stabilizing actually led to thermal destabilization of the proteins. The reasons for this contradiction between prediction and experiment were investigated. Conclusions The results reveal a mutational strategy to improve the biophysical behavior of nanobody binders and indicate a species-specificity of nanobody architecture.

Details

show
hide
Language(s):
 Dates: 2017-09
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: -
 Identifiers: DOI: 10.1016/j.bbagen.2017.06.014
BibTex Citekey: KunzFSZVSKMH2017
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Biochimica et Biophysica Acta: General Subjects
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 1861 (9) Sequence Number: - Start / End Page: 2196 - 2205 Identifier: -