Publication Date
2020
Document Type
Dissertation/Thesis
First Advisor
Horn, James R.
Degree Name
M.S. (Master of Science)
Legacy Department
Department of Chemistry and Biochemistry
Abstract
VHH domains are the antigen-binding units found in heavy-chain-only antibodies from the Camelidae family (e.g., camels and llamas). They are considered a unique minimalist antibody scaffold due to the lack of a variable light chain domain. Despite having half the antigen-binding interface of conventional antibodies, these VHH antibodies bind their target antigens with affinities comparable to conventional antibodies. This project explored how flexible linkers connecting an anti-GFP VHH (αGFP VHH) domain to its antigen (EGFP) enhanced antigen stability and influenced oligomeric assembly. It was hypothesized that linking the anti-GFP VHH antibody to its target antigen (EGFP) will increase the local concentration of antibody, thus increasing the stability of the antigen. Consequently, we wanted to test the effect of linker length on both thermal/chemical stability and oligomeric state. The results confirm that a flexible linker between the EGFP antigen and the anti-GFP VHH stabilized EGFP with an observed increase in melting temperature of 8-11°C. In addition to showing enhanced stability, the linker length affected the oligomeric assemblies of the EGFP-αGFP VHH fusions and induced the formation of monomers and higher-order oligomers EGFP. Since proteins are used in a wide range of applications, including medical imaging, therapeutics, and diagnostics, VHH fusions may serve as useful modules to extend the function of proteins across a wider range the environmental conditions (e.g., high temperature and harsh chemical conditions).
Recommended Citation
Mostafa, Nourhan Mohamed Disouky, "Single-Domain Vhh Antibodies as Protein Stabilization Modules" (2020). Graduate Research Theses & Dissertations. 7476.
https://huskiecommons.lib.niu.edu/allgraduate-thesesdissertations/7476
Extent
117 pages
Language
eng
Publisher
Northern Illinois University
Rights Statement
In Copyright
Rights Statement 2
NIU theses are protected by copyright. They may be viewed from Huskie Commons for any purpose, but reproduction or distribution in any format is prohibited without the written permission of the authors.
Media Type
Text
