Engineering and Kinetic Profiling of a Thrombin-Activable Prodrug Antibody Targeting P-Selectin

Clicks: 26
ID: 331535
2026
Free PDF
Article Quality & Performance Metrics
Overall Quality
Not rated
Combines reader engagement with the AI quality analysis. This article has not been analysed, so there is no overall score — reader engagement is measured and shown alongside.
AI Quality Assessment
Not analyzed
Readership in this journal
Emerging

Ranked #24 of 38 articles by views in Protein Engineering Design and Selection

Most read Least read

Bar heights use a square-root scale.

Mint this article as an NFT
Not yet minted

Create a permanent, verifiable on-chain record of this article on the Scimatic Network. The NFT is held in your Journament account, and you can withdraw it to your own wallet at any time.

5 SUSD one-off · no wallet required
Sign in to mint
Abstract
Antibody prodrugs represent a promising therapeutic format that provides advantages compared to conventional monoclonal antibody therapeutics. In this study, we engineered an antibody prodrug using an anti-P-selectin antibody as a model system, incorporating N-terminal heavy chain extensions comprising a linear P-selectin (CD62P) epitope for shielding, a thrombin-cleavable linker, and variable spacer in the linker. A prodrug antibody library was prepared comprising various N-terminal extensions of the heavy chain including an Ala-scan over the P-selectin epitope and the thrombin cleavage site sequence (50 variants in the library). The shielding library was thoroughly investigated for retained target binding kinetics and cleavage activation kinetics. Antibody prodrug variants achieving complete shielding of target binding to a completely open state were observed in the library (many orders of magnitude reduced binding based on surface plasmon resonance (SPR) kinetic measurement). Upon thrombin activation, the antibody prodrug variants obtained full target binding at low nM affinity. Thus, at such nM antibody prodrug concentrations, the liberated shielding extensions appeared to diffuse effectively away from the paratope upon cleavage based on SPR and fluorescence-activated cell sorting (FACS) analyses. Importantly, binding studies of variants of the linear epitope peptide alone showed many orders-of-difference in reduced binding compared to the fused extensions of the epitope sequence to the antibody prodrug. Comparison of the kinetic data with a structure model of the prodrug antibody reveals aspects of designing an optimal prodrug antibody that may have general applicability as a novel therapeutic format.
Reference Key
openalex_W7220378051 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Mie M. Mortensen, Susanne Nedergaard Grell, Thomas Egebjerg, Che Yang, Nikolai Lorenzen, Peter Kjær Kristensen, Jais Rose Bjelke
Are you one of these authors? Claim your profile
Journal Protein Engineering Design and Selection
Year 2026
DOI
10.1093/protein/gzag029
URL
https://doi.org/10.1093/protein/gzag029
Keywords Keywords not found

Citations

No citations found. To add a citation, contact the admin at info@scimatic.org

Comments

No comments yet. Be the first to comment on this article.