US 2022/0018852 A1 — Methods and Kits for Detecting Liver Dysfunction
Bibliographic Data
| Field | Value |
|---|---|
| PCT publication (primary reference) | WO 2020/104458 A1 |
| US publication | US 2022/0018852 A1 |
| European patent | EP 3 884 280 B1 (granted) |
| Spanish member | ES 2 971 966 T3 |
| US publication date | January 20, 2022 |
| PCT filing | PCT/EP2019/081801 — November 19, 2019 |
| Priority | EP 18306528.3 — November 20, 2018 |
| Inventors | Souleiman El Balkhi, Franck Saint-Marcoux, Jean-Baptiste Woillard (Limoges, FR) |
| Applicants | INSERM, Centre Hospitalier Régional Universitaire de Limoges, Université de Limoges |
| IPC | G01N 33/68 (2006.01) |
| CPC | G01N 33/6893; G01N 2333/765; G01N 2800/52; G01N 2800/50; G01N 2800/085 |
Title
“Methods and Kits for Detecting Liver Dysfunction in a Subject”
Abstract (verbatim excerpt)
Most chronic liver diseases are notoriously asymptomatic, until cirrhosis with clinical decompensation occurs. The use of early diagnosis strategies is vital to maintain patients in a symptom-free state and to delay decompensation. Albumin (HSA) undergoes several post-translational modifications in hepatocytes but clinical relevance of some of these modifications has been recently investigated in advanced liver diseases. Now, the inventors demonstrate that the binding capacities of some ligands, measured by inductively coupled plasma mass spectrometry (ICP-MS), are significantly different between cirrhotic patients and patients with no liver dysfunctions. The decreased binding capacities in cirrhotic patients were paralleled by the presence of significantly higher HSA isoforms. Animal experimentations were also conducted to explore the precocity of HSA modifications in the course of chronic liver dysfunction.
Core Innovation: The SEB Test
This patent covers the Serum Enhanced Binding (SEB) test — see SEB test for full methodology.
The test measures the functional binding capacity (effective binding capacity, eBC) of HSA in a serum sample by:
- Spiking serum ex vivo with defined quantities of specific ligands
- Separating albumin-bound vs. free fraction (ultrafiltration)
- Quantifying free (unbound) ligand by ICP-MS
Higher free fraction = albumin is structurally damaged = liver dysfunction.
Ligand Panel — 5 Binding Sites Covered
| Ligand | HSA Site | Clinical significance |
|---|---|---|
| Copper (Cu) | N-terminal site (NTS) | Most sensitive to ischemia and oxidative stress; elevated free Cu/HSA = strong cirrhosis marker |
| Cobalt (Co) | NTS (ACB site) | Historical ACB test (colorimetric); here measured with higher precision by ICP-MS |
| Gold (Au) | Cys34 (free thiol) | Free Au binding = intact Cys34 (HMA state); decreased Au binding = HNA1/HNA2 oxidation |
| Cadmium (Cd) | Multi-metal site A | Independent oxidative damage probe |
| L-Thyroxine (T4) + Dansylsarcosine | Sudlow Sites I & II (hydrophobic pockets) | Detects global conformational changes; tertiary structure destabilization |
Key Results (Patent Data)
- 18 cirrhotic vs. 18 controls: Free Cu/HSA and free Au/HSA significantly elevated in cirrhosis (Figures 2A, 2C)
- HSA isoform correlation: Elevated free ligands paralleled by higher HNA1/HNA2 and truncated isoforms (Figure 3)
- Rat DILI model: Detection of liver damage at D3 post-exposure vs. ALT/AST at D7 (4-day advantage)
- Specificity: L-thyroxine and dansylsarcosine changes indicate conformational (not just Cys34) damage
Relationship to Published Work
- el-balkhi-2024-seb — published validation of the SEB test in rat DILI model (Analytica Chimica Acta, 2024); this is the journal paper supporting the patent
- ALBOM study — clinical application of HSA isoform profiling (structural complement to SEB functional test)
Claims Summary
Primary claims cover:
- Method of detecting liver dysfunction by measuring unbound fraction of ≥1 metal ligand (Cu, Co, Au, Cd) spiked into a patient serum sample
- Use of ICP-MS for quantification of free metal fraction
- Panel of 5 ligands covering distinct albumin binding sites
- Kit for performing the assay (ligands + ICP-MS-compatible reagents)
- Extension to organic ligands (T4, dansylsarcosine) for Sudlow site assessment
Commercialization Angle
Primary licensing targets per technology-transfer:
- Grifols / CSL Behring — companion diagnostic for albumin infusion therapy (identify functionally dead albumin patients who need fresh albumin infusion)
- LabCorp / Mayo Clinic — specialty hepatic functional test in existing ICP-MS metals workflow
- ICON plc — Phase I safety (DILI early detection) in CRO specialty biomarker labs