Pharma & Biotech

Gene therapy

Gene therapy – creating safe and effective therapies

Gene therapy involves the transmission of genetic material into the patient's body cells to treat or potentially cure various genetic disorders.

Gene therapy is most commonly performed using viral-mediated gene transfer with Adeno-Associated Virus (AAV) or lentivirus. AAV vectors have a small packaging size (-5kb), but they are non-integrating viruses, offering prolonged transgene expression. They cannot propagate without helper viruses, which makes them safe for clinical use.

dPCR is also important in contamination control, which is an essential process in manufacturing environments aimed at minimizing and eliminating unwanted substances, particularly in the production of biopharmaceuticals. dPCR provides higher sensitivity of detection at a lower template input range, enabling more robust results.

Gene therapy workflow

Preparation

Plasmid preparation
and QC

In-process quality control

Virus vector titer and integrity

Product and process related impurities

Final product quality control

Viral vector titer, integrity and potency

Purity and safety

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AAV gene therapy | Viral titer quantification and integrity

Adeno-associated viruses (AAVs) play a crucial role in gene therapy as a delivery vehicle for therapeutic genes due to their ability to efficiently transduce a wide range of cell types and their relatively low immunogenicity. Accurate quantification and characterization of AAVs are critical for precise patient dosing. qPCR is widely used for AAV titer quantification. However, the technique is limited by the presence of PCR inhibitors that can hinder amplification and introduce variability, plus it requires a standard curve.

Conversely, digital PCR demonstrates improved amplification efficiency and higher tolerance to inhibitors than qPCR due to partitioning and end-point measurement without the need for a standard curve.

We have developed a new viral vector genome quantification workflow from capsid lysis to titer and integrity determination, including singleplex and multiplex assays for quantifying and qualifying AAVs using the QIAcuity Digital PCR System.

One of the benefits of using nanoplate dPCR for residual host cell DNA quantification is the easy setup and detection of host cell DNA thanks to a premixed master mix and positive/internal control. Another advantage is the accurate detection of E. coli, CHO and HEK293 residual DNA to low femtograms, even in the presence of PCR contaminants and inhibitors.

Partnership with Niba Labs

Niba Labs is committed to developing innovative analytical solutions for the biopharmaceutical industry focused on cell and gene therapies. We address quality attributes such as vector genome titer, vector genome integrity and biodistribution using state-of-the-art digital PCR techniques..

This World TB Day, we're partnering with Inter Medico to focus on the critical role of TB testing, especially in vulnerable groups and immigrants in Canada.

We can develop assays and perform sample testing for GLP-compliant biodistribution studies. Furthermore, we can support our customers from method development to method validation and sample testing as part of GMP certification for methods such as digital PCR, real-time PCR and ELISA. In addition, we support the characterization of samples through high-throughput sequencing and transmission electron microscopy. 

"Niba Labs is committed to developing innovative analytical solutions for the biopharma industry that focus on cell and gene therapies and thoroughly investigate quality attributes such as vector genome titer and vector genome integrity," said David Dobnik, PhD, CSO at Niba Labs.  

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Resources for gene therapy applications

Interested in a personalized package to meet your company’s needs?

We understand that one solution doesn’t fit all. We offer a range of products, technologies and services for gene therapy research. Contact us and together we can develop a package tailored for you.

Contamination control

Contamination control involves rigorous procedures and standards to ensure that contaminants, such as residual host-cell DNA (resDNA or HCD), do not compromise product safety and efficacy. Residual HCD carryover poses major safety concerns when manufacturing protein and vaccine therapeutics. Risks include oncogenicity, infectivity and immunogenicity caused by the final product. Therefore, levels of HCD must not exceed those established by regulatory agencies such as the U.S. Food and Drug Administration and the World Health Organization.

Moreover, contaminating DNA in your PCR reagents can be a significant source of background and false positives. Ultra Clean Production enhances the specificity and efficiency of probe-based digital PCR to provide accurate, singleplex or up to 5-plex analysis. Dedicated processes are implemented to enable Ultra Clean Production of the master mix, which minimizes contaminating DNA background and GC bias and gives you absolute certainty in your PCR reagents. This makes it an ideal choice for contamination-free, microbial applications as well as quality control applications, like residual DNA testing.

Mycoplasma Detection

Mycoplasmas are common contaminants in biological products derived from cell lines in the biopharmaceutical industry. They are often introduced either from the source cell lines or during production. Various guidelines and technical papers address the risks of mycoplasma contamination in the manufacturing process.

Digital PCR is an effective method for detecting these mycoplasma contaminations in cell cultures and related biological products. For instance, the QIAcuity Mycoplasma Quant Kit is a highly sensitive RT-dPCR kit that detects rRNA and DNA, covering 127 mycoplasma species. The kit includes an internal control to prevent false negatives caused by PCR inhibitors or errors in RNA extraction and RT reactions.

Meanwhile, the QIAcuity Residual DNA Quantification Kits provide accurate CHO, E.coli and HEK293 residual DNA (resDNA) quantification results with or without extraction, even in the presence of PCR contaminants or other inhibitory reagents. The assays are multicopy target assays that accurately determine highly fragmented residual host cell DNA.

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Accelerate your data-driven gene therapies

From data analysis to databases, from gene expression to genome assembly, from interpretation to integration – QIAGEN Digital Insights has the breadth and quality of bioinformatics tools you need to drive meaningful insights and fuel gene therapy discoveries.