NeoSickle and NeoHemog MALDI-MS platforms for sickle cell disease screening and hemoglobinopathy detection

Significance of the topic

Hemoglobinopathies, including sickle cell disease (SCD), are global public health priorities requiring reliable, high-throughput neonatal screening. Conventional workflows based on dried blood spots (DBS) and chromatographic separation such as HPLC deliver robust results but can be limited in throughput, standardization, and the spectrum of detectable variants. MALDI-MS approaches promise rapid acquisition, reproducibility, and scalability for large screening programs, offering potential improvements in laboratory efficiency and expanded variant detection.

Study objectives and overview

This study presents and evaluates two complementary MALDI-MS–based solutions from Shimadzu Chemistry & Diagnostics: NeoSickle, an IVDR-compliant intact-globin-chain platform tailored to SCD newborn screening, and NeoHemog, a peptide-based solution under development to broaden hemoglobin variant detection. Goals included demonstrating analytical concordance with established HPLC screening, assessing sensitivity and specificity for major variants (HbS, HbC, HbE), characterizing workflow automation and throughput, and exploring NeoHemog’s ability to flag rarer variants via peptide-specific alerts.

Methodology

Samples: Newborn DBS punches (3.2 mm) collected at approximately day 3 of life were used for NeoSickle protocols; additional adult and newborn cohorts were included in NeoHemog performance evaluation.

Sample preparation and workflow: NeoSickle uses an extraction solution to elute intact globin chains from a 3.2 mm DBS punch, mixes the extract with a matrix solution, and deposits the sample/matrix on MALDI targets. NeoHemog adds an enzymatic digestion step following elution to generate peptides prior to matrix addition and MALDI spotting. Automated processing on 96-well plates enables processing of large batches (reported ~90 minutes for 270 samples) and robotic workflows including puncher, liquid handling robot, and instrument integration.

Data handling and middleware: A LabGateway middleware interfaces kit batch management, instrument control, plate creation, raw-data traceability, quality control, and automatic transfer of validated results to external or national databases. eNeoSickle and eNeoHemog software provide visualization and biological result reporting.

Used instrumentation

The evaluated analytical platform was the MALDI-8020 EasyCare mass spectrometer. The workflow included NeoSickle kits (NS540/NS96), NeoHemog kits (NH360), automated puncher and liquid-handling robots, and LabGateway middleware for process and data management. NeoSickle has been implemented across multiple MALDI-MS systems in routine use.

Main results and discussion

NeoSickle performance: On a set of 787 samples compared with Variant NBS HPLC (Bio-Rad) across three HPLC systems, NeoSickle achieved near-complete concordance. Reported classification counts included the predominant FA pattern and FAS/FS patterns consistent with HPLC calls. For detection of samples carrying HbS (at least one HbS chain), NeoSickle sensitivity was 100% and specificity 99.85%; positive predictive value ~0.99 and negative predictive value 1. For outright SCD diagnosis, both sensitivity and specificity were 100% in the evaluated set.

NeoHemog performance: Preliminary evaluation on multiple adult and newborn cohorts showed high sensitivity and specificity for common variants. Reported ranges for sensitivity and specificity were approximately 93–100% depending on variant and cohort. Examples include HbS detection near 97–100% sensitivity/specificity across newborn and adult subsets, HbC and HbE detection with comparable high performance, and good discrimination of homozygous forms. NeoHemog also produced peptide-specific alerts that identified thirteen rare hemoglobin variants with consistently elevated target peptide scores compared to non-carrier samples, supporting feasibility for extended screening.

Throughput and deployment: NeoSickle has been operational in France for seven years across multiple MALDI-MS platforms, supporting >1.5 million sample analyses and deployed in four neonatal screening centers with combined throughput reported up to ~35,000 samples per month. Automated 96-well workflows and middleware integration support routine large-scale screening.

Benefits and practical applications

- High-throughput capability: automated 96-well protocols and rapid MALDI acquisition enable processing of hundreds to thousands of DBS samples with short turnaround times.
- Strong analytical performance: demonstrated high sensitivity and specificity for SCD and carrier detection, with excellent concordance to established HPLC methods.
- Scalability and standardization: kit-based reagents, middleware traceability, and cross-instrument deployment facilitate standardized workflows suitable for regional or national screening programs.
- Extended variant detection: peptide-based NeoHemog extends detection to additional and rare variants that intact-chain analysis may miss, enabling broader hemoglobinopathy surveillance and epidemiological studies.

Future trends and potential applications

- Expansion of peptide-target panels and machine-learning–assisted interpretation to increase the spectrum of identifiable variants and improve specificity for rare forms.
- Further automation and integration with laboratory information systems and national databases to streamline result validation and reporting.
- Wider adoption across newborn screening programs internationally, subject to regulatory approvals in respective jurisdictions.
- Combining intact-chain and peptide workflows in tiered screening strategies: rapid intact-chain screening (NeoSickle) for primary detection and peptide-based follow-up (NeoHemog) for variant clarification and expanded reporting.

Conclusions

NeoSickle delivers robust, IVDR-compliant intact-globin MALDI-MS screening for sickle cell disease with performance comparable to standard HPLC and demonstrated large-scale operational use. NeoHemog, in development, complements this capability by enabling peptide-based detection of a broader range of hemoglobin variants, including rare forms. Together these approaches support scalable, standardized neonatal screening workflows with potential to improve throughput and extend epidemiological monitoring of hemoglobinopathies. Regulatory considerations limit use in some regions (e.g., not FDA-cleared for diagnostic use in the United States), so local regulatory status should guide deployment.

References

• El Osta M., Gouriou Y., Naubourg P., Crevisy G., Devos C., Moreau S., Toyota Y., Ducoroy P. NeoSickle and NeoHemog MALDI-MS platforms for sickle cell disease screening and hemoglobinopathy detection. Poster WP-137. Shimadzu Chemistry & Diagnostics; Shimadzu Europa GmbH.