OAK

Enhanced Therapeutic Potential of Antibody Fragment by Prolonged in vivo Half-life via IEDDA mediated Albumin Conjugation

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Abstract
The application of single-chain variable fragments (scFvs) in treating human diseases, such as cancer and immune system disorders, has attracted significant attention. However, a critical drawback of scFv is its extremely short serum half-life, which limits its therapeutic potential. Thus, there is a critical need to prolong the serum half-life of the scFv for clinical applications.
To address this limitation, we coupled scFv with human serum albumin (HSA), the abundant protein in human blood. To minimize damage to the antigen-binding affinity of scFv upon the HSA conjugation, we employed a site-specific conjugation technique utilizing a heterobifunctional crosslinker that facilitates thiol-maleimide and inverse electron-demand Diels-Alder reactions (IEDDA). As a model protein, we selected 4D5scFv, derived from trastuzumab, a monoclonal antibody used in the treatment of human epithermal growth factor 2 (HER2)-positive breast cancer.
We introduced a tetrazine functional group (frTet) at conjugation site candidates of 4D5scFv predicted by computational methods. Using the linker TCO-PEG4-MAL, a single HSA molecule was site-specifically conjugated to the 4D5scFv (4D5scFv-HSA). The 4D5scFv-HSA conjugate exhibited anti-HER2 binding affinity similar to unmodified 4D5scFv. Furthermore, in mice, we noticed that the serum half-life of 4D5scFv-HSA was approximately 12 hours, a remarkable improvement as it was 85 times longer than that of the unmodified 4D5scFv.
The antigen binding results and pharmacokinetic profile of 4D5scFv-HSA demonstrate that the site-specific albumin-conjugated scFv retained its binding affinity with a prolonged serum half-life. In conclusion, we developed an effective strategy to prepare site-specific albumin-conjugated 4D5scFv, which can have versatile clinical applications with improved efficacy.
Author(s)
Eun Byeol, Go
Issued Date
2024
Type
Thesis
URI
https://scholar.gist.ac.kr/handle/local/19243
Alternative Author(s)
고은별
Department
대학원 신소재공학부
Advisor
Kwon, Inchan
Degree
Master
Appears in Collections:
Department of Materials Science and Engineering > 3. Theses(Master)
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