Characterization of a Phage Display-Selected scFv Against Neuropilin 1 (NRP1) Isolated from a Naïve Human Library.

Khadijeh Farrokhi, Fatemeh Nasiri, Pouya Safarzadeh Kozani, Pooria Safarzadeh Kozani

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Article in Molecular biotechnology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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4 authors.

Khadijeh FarrokhiDepartment of Microbial Biotechnology, Faculty of Biotechnology, Amol University of Special Modern Technologies, Amol, Iran.

Fatemeh NasiriDepartment of Internal Medicine, College of Medicine, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, MB, Canada. Fatima.Nasiri@umanitoba.ca.

Pouya Safarzadeh Kozani *Research and Development Center of Biotechnology, Tarbiat Modares University, Tehran, Iran. puyasafarzadeh@gmail.com.

Pooria Safarzadeh Kozani *Research and Development Center of Biotechnology, Tarbiat Modares University, Tehran, Iran. pooriasafarzadeh@modares.ac.ir.ORCID http://orcid.org/0000-0002-9901-7851

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Immune checkpoint inhibitors are established therapeutics for solid tumors. This study isolated a single-chain fragment variable (scFv) targeting neuropilin 1 (NRP1), a promising therapeutic target. A naïve human scFv phage display library underwent five biopanning rounds against NRP1. The lead clone (KZC28) was expressed and characterized. Binding affinity was quantified via ELISA. Specificity was assessed by cross-reactivity testing against structurally similar/unrelated antigens. Sensitivity determined the lower detection limit. Cell-based ELISA and flow cytometry evaluated binding to endogenous NRP1 on U87-MG glioblastoma and PC3 prostate cancer cells. Computational modeling predicted structural stability, solubility, flexibility, and binding energetics, focusing on complementarity-determining region (CDR) interactions. KZC28 exhibited high affinity (K

Indexed as

Neuropilin-1Single-Chain AntibodiesCell Line, TumorCell Surface Display TechniquesComplementarity Determining RegionsEnzyme-Linked Immunosorbent AssayHumansPeptide LibraryProtein BindingComplementarity Determining RegionsNeuropilin-1NRP1 protein, humanPeptide LibrarySingle-Chain AntibodiesCancer immunotherapyMonoclonal antibodyNeuropilin 1Phage displaySingle-chain fragment variable

Identifiers

PMID40707855

DOI10.1007/s12033-025-01480-y

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.