Redefining Plasma Proteomics: How Orbitrap Astral and Olink Reveal Address the Coverage Gap
The Plasma Proteomics Challenge
Plasma is one of the most information‑rich biospecimens — circulating proteins reflect the state of nearly every tissue and organ. But it is also one of the most analytically challenging matrices.
The core difficulty is the dynamic range problem: plasma protein concentrations span over 10 orders of magnitude, from abundant albumin and immunoglobulins to ultra‑low‑abundance cytokines and signaling molecules measured in fg/mL [1]. No single platform can capture this full spectrum.
Historically, researchers faced a binary choice: broad but shallow (untargeted mass spectrometry identifying thousands of proteins but missing the rarest signals) or narrow but deep (targeted immunoassays quantifying a predefined panel with exquisite sensitivity). The Orbitrap Astral and Olink Reveal represent the two most advanced answers to this dilemma — and as we show below, combining them breaks the trade‑off entirely.
Orbitrap Astral: Unbiased Depth at Unprecedented Speed
Introduced in 2023, the Orbitrap Astral combines the proven Orbitrap analyzer with a high‑speed Astral analyzer, achieving high resolution, sensitivity, and acquisition rates up to 200 Hz simultaneously [1].
Notably, Novogene leverages nanoparticle (NP)‑based enrichment combined with the Astral platform to achieve 6,000+ protein identifications per sample at a throughput of 60 samples per day (SPD) — bridging the gap between deep proteome coverage and high‑throughput demands for large‑scale cohort studies.
Measurement data of plasma/serum samples and technical replicate correlation analysis
Olink Reveal: Targeted Precision at Scale
Olink Reveal is Olink’s newly launched product, built on PEA technology with NGS readout to quantify 1,000+ curated proteins from just 4 μL of sample [2].
What Makes Reveal Different
| Feature | Olink Reveal |
| Protein Panel | 1,000+ proteins, robustly detectable markers |
| Immune Focus | 537 inflammation proteins covering 96% of immune response pathways in Reactome |
| Pathway Coverage | 100% of Reactome top‑level pathways; 64% of all pathway levels |
| Sensitivity | Detects as low as fg/mL; dynamic range spanning 10 logs |
| Sample Input | 4 μL serum or plasma; no high‑abundance protein depletion required |
| Precision | Intra‑plate CVs 8.1%; inter‑plate CVs 4.8%; inter‑site CVs 6.3% |
| Genetic Validation | 850+ proteins confirmed as disease‑relevant in UK Biobank; 700+ with cis‑pQTL associations |
| Readout | NGS‑based; normalized NPX values |
| Sample Types | Plasma, serum, CSF, urine, tissue lysates, cell culture supernatant, and more |
Why Combine Astral MS with Olink Reveal?
Based on our internal test data, four complementary strengths make the case for combining these platforms.
1. Abundance Distribution: Complementary Dynamic Ranges
Based on HPPP database abundance annotations [4], MS and Olink operate at fundamentally different concentration ranges. MS‑detected proteins have a median of 0.74 ng/mL (28.3% at ≥10 ng/mL), capturing the high‑abundance proteome. Olink‑detected proteins center at 0.26 ng/mL (64.8% between 0.01–1 ng/mL), reaching the low‑abundance signaling proteome. Together they span 11+ orders of magnitude.
Concentration coverage: MS captures the high‑abundance proteome; Olink reaches the low‑abundance signaling proteome. Together spanning 11+ orders of magnitude.
2. Proteome Coverage: Expanded Detection Breadth
MS profiles thousands of proteins unbiased, while Olink targets 1,000+ proteins at sub‑pg/mL sensitivity [1][2]. Each platform detects proteins the other misses — MS captures intracellular and membrane proteins invisible to targeted panels; Olink reaches low‑abundance cytokines below MS limits. Combining both dramatically expands the detectable proteome.
Protein overlap analysis: each platform detects unique proteins; combining dramatically expands coverage. HPPP (4,608 proteins) as reference baseline.
3. Orthogonal Cross‑Validation: Platform Concordance
Proteins detected by both platforms form a high‑confidence core proteome. Over half of Olink’s panel is independently confirmed by MS, with a robust core validated across MS, Olink, and the HPPP reference [4]. These shared proteins are enriched in vesicle transport, immune regulation, and cell adhesion. Dual‑platform cross‑verification significantly reduces false discovery rates.
Cross‑validation: shared core in GO:CC, KEGG, and Reactome.
4. Functional Complementarity: Distinct Enrichment Profiles
Functional enrichment analysis of all proteins detected by each platform reveals distinct biological profiles [1][2]. MS‑identified proteins are enriched in core cellular processes — Translation, Vesicle Transport, and RNA Binding. Olink‑identified proteins concentrate in immune and signaling biology — Cytokine Signaling, Inflammatory Response, and Receptor‑Ligand Activity. KEGG pathway analysis further highlights MS‑enriched Endocytosis and neurodegenerative pathways, while Olink‑enriched JAK‑STAT, NF‑κB, IL‑17, and PI3K‑Akt signaling. Together they span from intracellular machinery to extracellular immune coordination.
MS top 5: Translation, Vesicle Transport, RNA Binding (cellular machinery). Olink top 5: Cytokine Signaling, Inflammatory Response, Receptor‑Ligand Activity (immune signaling). Analysis based on all proteins detected by each platform.
Published Evidence
Yang et al. [2] (Commun. Biol., 2022): Olink screening → MRM‑MS validation identified a 4‑protein AMS signature (AUC = 0.9).
Schiff et al. [3] (JCI Insight, 2024): MS discovery → Olink validation yielded a 6‑protein TB panel (FETUB, FCGR3B, LRG1, SELL, CD14, ADA2; AUC = 0.943), exceeding WHO specificity targets.
Boucherat et al. [1] (Nat. Cardiovasc. Res., 2022): Combined tissue MS, transcriptomics, and plasma Olink to identify LTBP‑2 as a PAH biomarker — multi‑platform concordance strengthened clinical validation.
Key Takeaway: MS provides unbiased proteome‑wide coverage; Olink Reveal delivers ultra‑sensitive targeted quantification. Published studies consistently confirm their complementary value [1][2][3] — the combined workflow is the most powerful approach for modern plasma proteomics.
Why Choose Novogene?
1. Ultra‑Deep Proteome Coverage Combined LC‑MS/MS and Olink workflows deliver reliable identification and broader coverage of low‑abundance circulating proteins — maximizing the biological insights you can extract from every sample.
2. High Daily Throughput Complete LC‑MS/MS and bioinformatics analysis within 30 business days (n ≤ 50), shortening your project turnaround time without compromising data quality.
3. Trusted Orbitrap Astral Platform High‑precision mass spec instrumentation delivers stable, repeatable, publication‑grade quantitative datasets suitable for clinical and translational research.
4. Certified Olink CSP Provider As an official Olink Certified Service Provider with a US‑based lab, we ensure quality‑assured local delivery and full compliance with Olink’s rigorous standards.
5. End‑to‑End Workflow From sample preparation through LC‑MS/MS analysis to expert data interpretation, we deliver a fully integrated proteomics solution — so you can focus on the science.
References
1. Boucherat O, Yokokawa T, Krishna V, et al. Identification of LTBP‑2 as a plasma biomarker for right ventricular dysfunction in human pulmonary arterial hypertension. Nat. Cardiovasc. Res., 2022, 1(8): 748–760.
2. Yang J, Jia Z, Song X, et al. Proteomic and clinical biomarkers for acute mountain sickness in a longitudinal cohort. Commun. Biol., 2022, 5(1): 548.
3. Schiff HF, Garay‑Baquero DJ, White CH, et al. Comprehensive plasma proteomic profiling reveals biomarkers for active tuberculosis. JCI Insight, 2024, 9(8): e173273.
4. Omenn GS, Lane L, Lundberg EK, et al. The Human Plasma Proteome Project (HPPP): Challenges and Opportunities. Mol. Cell. Proteomics, 2021, 20: 100062.