---
title: "Peptide-based digital ELISA on single-molecule arrays for ultrasensitive endotoxin detection"
id: "pubmed-42663432"
canonical_url: "https://medichelpline.com/clinical-feed/pubmed-42663432"
content_type: "clinical_feed_article"
specialty: "Critical Care"
source_name: "PubMed / NCBI"
source_url: "https://pubmed.ncbi.nlm.nih.gov/42663432/"
doi: "10.1039/d6an00937a"
published_at: "2026-08-28T00:00:00.000Z"
evidence_level: "Journal Article"
license: "CC-BY-NC-4.0 / Informational Use"
---
# Peptide-based digital ELISA on single-molecule arrays for ultrasensitive endotoxin detection
## Provenance & Clinical Metadata
- **Canonical URL:** https://medichelpline.com/clinical-feed/pubmed-42663432
- **Specialty:** [Critical Care](https://medichelpline.com/clinical-feed/critical-care.md)
- **Primary Source:** PubMed / NCBI
- **Source URL:** [Original Journal Publication](https://pubmed.ncbi.nlm.nih.gov/42663432/)
- **DOI:** [10.1039/d6an00937a](https://doi.org/10.1039%2Fd6an00937a)
- **Published At:** 2026-08-28T00:00:00.000Z
- **Evidence Rating:** Journal Article
## Executive GIST (TL;DR)
- Endotoxin detection is essential for sepsis diagnosis and pharmaceutical safety. The source describes a **peptide-based digital ELISA (dELISA)** that uses two high-affinity, phage-displayed peptides on a single-molecule array platform to detect endotoxin without antibodies. - Magnetic bead surface chemistry was optimized by **EDC/NHS** carboxyl activation at 10 °C, and **L-lysine (Lys)** was selected as a blocking agent to preserve bead monodispersity via electrostatic repulsion. - Under optimized conditions the assay achieved a limit of detection of **0.2 pg mL-1** and a linear dynamic range of **0–200 pg mL-1** for endotoxin. - The method showed strong analytical performance: coefficient of variation **< 15%**, and tested interference resistance with recovery rates of **76.9%–110.1%** in pharmaceutical formulations including saline, 5% dextrose, and commercial ophthalmic solutions. - Clinical validation included quantification of endotoxin in plasma from septic and non-septic patients and demonstrated **78.0%** recovery in whole blood spiked with exogenous endotoxin. - By replacing antibodies with peptide binders and integrating single-molecule arrays, the platform aims to overcome antibody-related limitations and offer a robust, translational tool for direct endotoxin measurement in complex biological matrices and pharmaceutical products.
## Clinical Analysis & Structured Key Points
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Online ahead of print. # Peptide-based digital ELISA with single-molecule arrays for ultrasensitive and specific endotoxin detection in clinical plasma and pharmaceutical products [Junrong Li](https://pubmed.ncbi.nlm.nih.gov/?term=Li+J&cauthor_id=42663432)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42663432/#full-view-affiliation-1 "Department of Pharmaceutical Sciences, State Key Laboratory of Fine Chemicals, School of Chemical Engineering, Dalian University of Technology, Dalian, Liaoning 116081, China."), [Kun Cao](https://pubmed.ncbi.nlm.nih.gov/?term=Cao+K&cauthor_id=42663432)[ 2 ](https://pubmed.ncbi.nlm.nih.gov/42663432/#full-view-affiliation-2 "Dalian Third People's Hospital, Dalian, Liaoning 116033, China, \(Affiliated to Dalian University of Technology\)."), [Heqing Yang](https://pubmed.ncbi.nlm.nih.gov/?term=Yang+H&cauthor_id=42663432)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42663432/#full-view-affiliation-1 "Department of Pharmaceutical Sciences, State Key Laboratory of Fine Chemicals, School of Chemical Engineering, Dalian University of Technology, Dalian, Liaoning 116081, China."), [Jicheng Dong](https://pubmed.ncbi.nlm.nih.gov/?term=Dong+J&cauthor_id=42663432)[ 2 ](https://pubmed.ncbi.nlm.nih.gov/42663432/#full-view-affiliation-2 "Dalian Third People's Hospital, Dalian, Liaoning 116033, China, \(Affiliated to Dalian University of Technology\).")[ 3 ](https://pubmed.ncbi.nlm.nih.gov/42663432/#full-view-affiliation-3 "Faculty of Medicine, Dalian University of Technology, Dalian, Liaoning 116024, China. dongjicheng@dlut.edu.cn."), [Quanwu Wang](https://pubmed.ncbi.nlm.nih.gov/?term=Wang+Q&cauthor_id=42663432)[ 2 ](https://pubmed.ncbi.nlm.nih.gov/42663432/#full-view-affiliation-2 "Dalian Third People's Hospital, Dalian, Liaoning 116033, China, \(Affiliated to Dalian University of Technology\)."), [Feng Hai](https://pubmed.ncbi.nlm.nih.gov/?term=Hai+F&cauthor_id=42663432)[ 2 ](https://pubmed.ncbi.nlm.nih.gov/42663432/#full-view-affiliation-2 "Dalian Third People's Hospital, Dalian, Liaoning 116033, China, \(Affiliated to Dalian University of Technology\).") Affiliations Expand ### Affiliations * 1 Department of Pharmaceutical Sciences, State Key Laboratory of Fine Chemicals, School of Chemical Engineering, Dalian University of Technology, Dalian, Liaoning 116081, China. * 2 Dalian Third People's Hospital, Dalian, Liaoning 116033, China, (Affiliated to Dalian University of Technology). * 3 Faculty of Medicine, Dalian University of Technology, Dalian, Liaoning 116024, China. dongjicheng@dlut.edu.cn. * PMID: **42663432** * DOI: [ 10.1039/d6an00937a ](https://doi.org/10.1039/d6an00937a) Item in Clipboard # Peptide-based digital ELISA with single-molecule arrays for ultrasensitive and specific endotoxin detection in clinical plasma and pharmaceutical products Junrong Li et al. Analyst. 2026. Show details Display options Display options Format Abstract PubMed PMID Analyst Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Analyst%22%5Bjour%5D&sort=date&sort_order=desc) * [ Search in NLM Catalog ](https://www.ncbi.nlm.nih.gov/nlmcatalog?term=%22Analyst%22%5BTitle+Abbreviation%5D) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42663432/) . 2026 Aug 28. doi: 10.1039/d6an00937a. Online ahead of print. ### Authors [Junrong Li](https://pubmed.ncbi.nlm.nih.gov/?term=Li+J&cauthor_id=42663432)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42663432/#short-view-affiliation-1 "Department of Pharmaceutical Sciences, State Key Laboratory of Fine Chemicals, School of Chemical Engineering, Dalian University of Technology, Dalian, Liaoning 116081, China."), [Kun Cao](https://pubmed.ncbi.nlm.nih.gov/?term=Cao+K&cauthor_id=42663432)[ 2 ](https://pubmed.ncbi.nlm.nih.gov/42663432/#short-view-affiliation-2 "Dalian Third People's Hospital, Dalian, Liaoning 116033, China, \(Affiliated to Dalian University of Technology\)."), [Heqing Yang](https://pubmed.ncbi.nlm.nih.gov/?term=Yang+H&cauthor_id=42663432)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42663432/#short-view-affiliation-1 "Department of Pharmaceutical Sciences, State Key Laboratory of Fine Chemicals, School of Chemical Engineering, Dalian University of Technology, Dalian, Liaoning 116081, China."), [Jicheng Dong](https://pubmed.ncbi.nlm.nih.gov/?term=Dong+J&cauthor_id=42663432)[ 2 ](https://pubmed.ncbi.nlm.nih.gov/42663432/#short-view-affiliation-2 "Dalian Third People's Hospital, Dalian, Liaoning 116033, China, \(Affiliated to Dalian University of Technology\).")[ 3 ](https://pubmed.ncbi.nlm.nih.gov/42663432/#short-view-affiliation-3 "Faculty of Medicine, Dalian University of Technology, Dalian, Liaoning 116024, China. dongjicheng@dlut.edu.cn."), [Quanwu Wang](https://pubmed.ncbi.nlm.nih.gov/?term=Wang+Q&cauthor_id=42663432)[ 2 ](https://pubmed.ncbi.nlm.nih.gov/42663432/#short-view-affiliation-2 "Dalian Third People's Hospital, Dalian, Liaoning 116033, China, \(Affiliated to Dalian University of Technology\)."), [Feng Hai](https://pubmed.ncbi.nlm.nih.gov/?term=Hai+F&cauthor_id=42663432)[ 2 ](https://pubmed.ncbi.nlm.nih.gov/42663432/#short-view-affiliation-2 "Dalian Third People's Hospital, Dalian, Liaoning 116033, China, \(Affiliated to Dalian University of Technology\).") ### Affiliations * 1 Department of Pharmaceutical Sciences, State Key Laboratory of Fine Chemicals, School of Chemical Engineering, Dalian University of Technology, Dalian, Liaoning 116081, China. * 2 Dalian Third People's Hospital, Dalian, Liaoning 116033, China, (Affiliated to Dalian University of Technology). * 3 Faculty of Medicine, Dalian University of Technology, Dalian, Liaoning 116024, China. dongjicheng@dlut.edu.cn. * PMID: **42663432** * DOI: [ 10.1039/d6an00937a ](https://doi.org/10.1039/d6an00937a) Item in Clipboard Cite Display options Display options Format Abstract PubMed PMID ## Abstract Endotoxin detection is critical for the diagnosis and management of sepsis and for the safety evaluation of pharmaceutical products. Here, we report a peptide-based digital enzyme-linked immunosorbent assay (dELISA) that integrates two high-affinity, phage-displayed peptides into a single-molecule array platform, enabling highly sensitive and specific endotoxin detection without the use of antibodies. The carboxylated activation of magnetic beads was optimized using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and _N_ -hydroxysuccinimide (NHS) at 10 °C, and L-lysine (Lys) was identified as an effective blocking agent that maintained bead monodispersity through electrostatic repulsion. Under optimized conditions, the assay achieved a detection limit of 0.2 pg mL-1 with a linear range spanning 0-200 pg mL-1. The method exhibited strong accuracy (coefficient of variation (CV) < 15%) and interference resistance (recovery rates of 76.9%-110.1%) when applied to pharmaceutical formulations such as saline, 5% dextrose, and commercial ophthalmic solutions. In clinical validation, the platform successfully quantified endotoxin in plasma samples from septic and non-septic patients and demonstrated excellent recovery (78.0%) in whole blood spiked with exogenous endotoxin. By eliminating antibody-related limitations, this peptide-based dELISA offers a robust and translational tool for direct endotoxin detection in complex biological matrices. 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