Cell and gene therapy:
Precision characterization for advanced biologics

The transformative potential of Cell and Gene Therapy (CGT) hinges on the meticulous characterization and quality control of advanced biologics. At OHMX.bio, we provide a comprehensive suite of expertise to accelerate your CGT development, ensuring the safety, efficacy, and consistency of your therapeutic products. Leveraging cutting-edge sequencing solutions and advanced bioinformatics, we empower researchers and clinicians with unparalleled insights into their cell lines, viral vectors, and mRNA constructs.
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Navigating the complexities of cell and gene therapy with OHMX.bio

The landscape of biologics, from engineered cell lines to viral vectors and mRNA constructs, demands rigorous quality control at every stage of development and manufacturing. Traditional QC methods often fall short in providing the precise, in-depth genetic and epigenetic insights required for these complex therapeutics. OHMX.bio bridges this gap by applying state-of-the-art sequencing, particularly Oxford Nanopore long-read sequencing, as a central tool for the quality control of biologics. This platform allows for complete and highly accurate sequence analysis of complex biological products, including mRNA constructs, plasmids, viral genomes, and engineered cell lines. Long-read sequencing enables the direct detection of full-length molecules, structural variants, and epigenetic modifications in a single run, providing deeper insights than conventional short-read or PCR-based methods.
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Core expertise in cell line characterization

Ensuring the genomic integrity of engineered cell lines is a fundamental step in the development of biologics. OHMX.bio specializes in applying long-read sequencing to provide the most comprehensive and accurate picture of cell line quality. Our approach allows for the full characterization of engineered modifications, detection of unintended edits, and precise mapping of insertions or transgenes.
Our approach allows for:

Detection of full-length insertions and their exact genomic locations

Traditional short-read methods often miss large structural variations or cannot resolve full insertion sequences. By contrast, OHMX.bio uses Oxford Nanopore long-read sequencing to directly read entire constructs and flanking genomic regions. This enables the detection of full-length insertions and their exact genomic locations.

Identification of rearrangements, duplications, or deletions

This level of detail is crucial for confirming the identity, purity, and stability of production cell lines used in biologics manufacturing.

Verification of on-target vs. off-target integration

Genomic profiling enables researchers to identify genetic variations, structural changes, and mutations that underpin disease mechanisms. The same profiling techniques can be used to validate genetic edits in cell and gene therapy applications, ensuring their safety and efficacy.

Confirmation of engineered regions and monitoring of genetic drift over time

OHMX.bio’s workflows include dedicated protocols for sample preparation and sequencing of various cell types, including CHO, HEK293, and other mammalian or even plant-derived systems. Our platform provides end-to-end insight into the confirmation of engineered regions, detection of residual plasmid sequences, and monitoring of genetic drift over time.

All sequencing outputs are processed through OHMX.bio’s validated bioinformatics pipelines, ensuring accurate variant calling, insertion mapping, and structural analysis. Our team delivers publication-ready reports and works with clients to interpret complex data in the context of quality control. This makes our service ideal for both early development and late-phase validation of engineered cell lines.
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Case study: Precise transfected cell line
QC with long-read sequencing

A critical challenge in cell and gene therapy is the accurate characterization of gene insertions within transfected cell lines. Traditional methods often provide limited information on insertion sites and the complete structure of the inserted genetic material.

In a real-world case study, OHMX.bio demonstrated the power of long-read sequencing for the quality control of a transfected human cell line where a custom reporter block was introduced at unknown genomic locations. Despite the exact structure of the reporter block being initially unknown, beyond the promoter sequence and the presence of a reporter gene and transcriptional response elements, OHMX.bio’s approach provided a comprehensive solution.

Libraries were prepared from the modified human cells and sequenced on a PromethION R10.4.1 flow cell, yielding an overall genome coverage of around 52X, sufficient for consensus sequence determination. By performing structural variant calling and filtering for homology with the known promoter sequence, a rough assembly of the reporter block could be constructed.

The key advantage of long-read sequencing in this study was its ability to cover the full reporter block and capture extensive information about the neighboring genomic regions where the reporter block was inserted. This enabled the precise determination of 13 distinct genomic insertion events at a base-specific level. This number of insertions was further confirmed by calculating the ratio between the read coverage over the reporter and the overall genome coverage.

Furthermore, the long reads allowed the assembly of a detailed nucleotide sequence of the reporter block itself. This facilitated the determination of the exact number of transcriptional response elements, the specific type of reporter gene, and the precise location of all these elements within the reporter block. This case study highlights how OHMX.bio’s custom in-house developed pipeline, combined with long-read sequencing, provides accurate localization of integration sites and reveals the full reporter sequence structure, addressing crucial quality control questions in cell line characterization.
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Partner with OHMX.bio for your cell and gene therapy needs

OHMX.bio combines deep scientific expertise with cutting-edge long-read sequencing technology to deliver actionable insights for your cell and gene therapy programs. Our commitment to scientific rigor, personalized collaboration, and fast turnaround times ensures that you receive the highest quality insights precisely when you need them.

From initial cell line engineering to preclinical and clinical use, OHMX.bio brings years of sequencing and analytical experience to support your QC strategy. Contact us today to discuss how our expertise in cell line characterization and broader Cell and Gene Therapy solutions can support your breakthrough research and development.
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