What is Olink Explore HT? A Comprehensive Analysis of the High-Throughput Protein Analysis Platform
In precision medicine, drug discovery, and large-scale cohort investigations, the ability to efficiently quantify thousands of proteins from minimal sample volumes represents a central challenge in modern proteomics research. Leveraging Proximity Extension Assay (PEA) technology, Olink has developed the Explore HT (High Throughput) platform, offering a next-generation, fully automated solution for high-throughput protein analysis. Relative to the earlier Explore platform, Explore HT delivers substantial advancements in throughput, analytical speed, data consistency, and scalability for large projects. This study provides a detailed overview of the platform’s technical principles, performance characteristics, application scenarios, and data analysis workflow, and further describes how MtoZ Biolabs supports research teams in effectively deploying Olink Explore HT to accelerate scientific progress.
Technical Principles of Olink Explore HT
1. Core Technology: Proximity Extension Assay (PEA)
(1) The assay is based on paired antibody probes that simultaneously recognize the target protein, with each probe conjugated to a unique oligonucleotide tag.
(2) Upon dual binding to the same protein, the oligonucleotides are brought into close proximity, enabling DNA polymerization to generate an amplifiable template.
(3) The resulting signals are quantified using next-generation sequencing (NGS), allowing highly sensitive and multiplex protein detection.
2. Key Advancements in the Olink Explore HT Platform
(1) Full Automation: Incorporates liquid-handling robotics and standardized plate-based workflows to minimize manual intervention.
(2) Ultra-High Throughput: Processes thousands of samples per batch while simultaneously profiling over 3,000 proteins.
(3) Enhanced Batch Consistency: Achieves inter-plate coefficients of variation (CV) typically <10%, enabling robust longitudinal and multi-center studies.
(4) Minimal Sample Requirement: Requires only 1–3 μL of plasma or serum per sample.
Performance Advantages of Olink Explore HT
1. Exceptional Throughput and Analytical Speed
(1) Achieves approximately a two- to three-fold increase in detection speed compared with the Explore platform.
(2) Supports high-throughput processing of tens of thousands of samples, making it particularly suitable for large epidemiological cohorts and geographically distributed clinical studies.
2. High Sensitivity and Broad Dynamic Range
(1) Detection limits reach the femtogram-per-milliliter (fg/mL) range, spanning approximately 10 orders of magnitude in protein concentration.
(2) Demonstrates superior capability for detecting low-abundance analytes, including cytokines and hormones, relative to most conventional mass spectrometry approaches.
3. Standardized Data Output: NPX Values
(1) All measurements are reported as standardized NPX (Normalized Protein eXpression) values, calculated using the Olink proprietary algorithm.
(2) NPX values represent relative protein abundance on a log₂ scale, allowing direct comparison across samples and batches.
(3) When combined with reference standards, NPX values can be converted to absolute concentrations, supporting the needs of clinical research.
4. Reduced Project Cost and Manual Labor
(1) Automated workflows minimize human error and improve data reproducibility.
(2) For large-scale projects, per-sample assay costs are significantly lower compared with traditional multi-batch manual workflows.
Application Scenarios of Explore HT
1. Large-Scale Clinical Cohort Studies
(1) Enables parallel quantification of thousands of plasma or serum samples.
(2) Suitable for epidemiological studies, disease risk prediction, and biomarker discovery.
2. Drug Development and Clinical Translation
(1) Facilitates pharmacodynamic assessments and toxicology biomarker screening.
(2) Assists pharmaceutical developers in rapidly validating drug targets and safety parameters during clinical trials.
3. Longitudinal Multi-Center Projects
(1) Standardized workflows ensure inter-center data comparability.
(2) Supports integration with transcriptomic and metabolomic datasets, enabling accelerated multi-omics analyses.
Data Analysis Workflow and Interpretation
1. Data Generation and Preprocessing
(1) NGS generates tag count data, with blank wells and anomalous signals removed during quality control.
(2) Data are normalized using internal control proteins and amplification controls to mitigate batch effects.
2. NPX Calculation and Statistical Analysis
(1) NPX values are reported on a log₂ scale, typically ranging from 5 to 15.
(2) Group comparisons are commonly performed using t-tests, ANOVA, and false discovery rate (FDR) correction, with fold change calculated as 2^(ΔNPX) to assess differential expression.
3. Downstream Bioinformatics Analysis
(1) Differentially expressed proteins undergo pathway enrichment analysis using KEGG and Reactome databases.
(2) Multi-omics integration with transcriptomics and metabolomics is employed to elucidate mechanistic networks.
(3) Results are visualized through heatmaps, volcano plots, and hierarchical clustering to provide an intuitive interpretation of findings.
As a next-generation high-throughput protein analysis platform, Olink Explore HT combines automation, exceptional throughput, high sensitivity, and standardized data outputs, making it highly applicable to large-scale cohort studies, drug development, and multi-center clinical trials. Through its advanced Olink platform, specialized bioinformatics expertise, and integrated multi-technology solutions, MtoZ Biolabs delivers end-to-end support, from experimental design to data interpretation, and from biomarker discovery to validation, thereby facilitating the rapid translation of research findings into clinical and industrial applications.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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