- Article
Generation of an Insecticidal Human Domain Antibody from a Phage Library Targeting Plutella xylostella Brush-Border Membrane Vesicles
- Xiaodan Hu,
- Xiao Zhang and
- Chunqin Xie
- + 11 authors
The importance of protein-based materials in agricultural pest control has received increasing attention in recent years. Herein, Plutella xylostella brush-border membrane vesicles (BBMVs) were used as a target to screen for human domain antibodies with insecticidal activity. Three rounds of panning of the phage display library yielded the domain antibody C4D, which competed with the Cry1Ac toxin to bind to P. xylostella BBMVs. Against P. xylostella larvae, the recombinant soluble C4D protein showed an LC50 of 1.57 μg/cm2 (95% fiducial limits: 0.83–2.54). Using pull-down assays and liquid chromatography–tandem mass spectrometry, we identified the C4D binding partner in P. xylostella midgut BBMVs to be a cadherin-like protein. Bio-Layer Interferometry assay revealed that the dissociation constant between soluble C4D and P. xylostella cadherin-like protein was 2.99 × 10−6 M. Thus, the present study explored strategies to generate insecticidal antibodies, and the human domain antibody C4D identified and characterized in this study can serve as a framework for generating novel insecticidal agents.
8 February 2026




![Impact of red seaweed-derived MAAs on redox balance and disease risk. The scheme provides a simplified representation of the complex processes and mediators involved. Environmental stressors include α-particles emitted during radon (Rn) decay and Escherichia coli. Mitochondria, through electron transport chain (ETC), generate reactive oxygen species (ROS) as they reduce O2 to H2O while producing energy in the form of ATP [21,22]. Although mitochondria are a major source, ROS are also produced in peroxisomes and by cytoplasmatic enzymes like NOX. Environmental stressors further exacerbate ROS formation. Key ROS include the superoxide radical (O2•−), hydrogen peroxide (H2O2) and the hydroxyl radical (•OH). The latter is formed through the Haber–Weiss reaction (interaction of O2•− with H2O2) and the Fenton reaction mediated by heavy metals such as Fe2+ [23,24]. When endogenous enzymatic (e.g., superoxide dismutase (SOD), catalase (CAT), glutathione peroxidases (GPXs)) and non-enzymatic antioxidants (e.g., glutathione (GSH)) are supported with the neutralizing capacity of dietary antioxidants like MAAs, the capability of the organism to prevent oxidative stress increases [21,25,26]. As ROS and environmental stressors activate macrophages though the nuclear factor kappa B (NF-κB) signaling pathway, inflammation occurs [27]. NF-κB upregulates inducible nitric oxide synthase (iNOS), catalyzing the conversion of L-arginine into L-citrulline and releasing nitric oxide (NO). This process generates further reactive nitrogen species (RNS), such as peroxynitrite (ONOO−). Simultaneously, pro-inflammatory cytokines, including tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) are secreted [6,27]. Macrophages also contribute to ROS production via NOX during respiratory bursts [20]. However, the broad arsenal of antioxidants can neutralize excessive levels of ROS and RNS through antioxidant and anti-inflammatory actions, thereby preventing chronic inflammation and the intertwined effects of oxidative and nitrosative stress, which would otherwise reinforce each other in a vicious cycle, leading to progressive damage to biomolecules [20,27,28,29,30]. Consequently, these compounds may ultimately contribute to a reduced risk of associated disease development and/or progression [21]. Abbreviations: ETC, electron transport chain; NOX, nicotinamide adenine dinucleotide phosphate oxidases; ATP, adenosine triphosphate; ROS, reactive oxygen species; O2•−, superoxide radical; H2O2, hydrogen peroxide; •OH, hydroxyl radical; SOD, superoxide dismutase; CAT, catalase; GPXs, glutathione peroxidases; MAAs, mycosporine-like amino acids; GSH, glutathione; NF-κB, nuclear factor kappa B; iNOS, inducible nitric oxide synthase; NO, nitric oxide; RNS, reactive nitrogen species; ONOO−, peroxynitrite; TNF-α, tumor necrosis factor-alpha; IL-6, interleukin-6.](https://mdpi-res.com/cdn-cgi/image/w=281,h=192/https://mdpi-res.com/cimb/cimb-48-00190/article_deploy/html/images/cimb-48-00190-g001-550.jpg)


