With over 20 years of molecular bioanalytical experience and industry thought leadership insights, we can help you advance even your most challenging projects when it matters most. Located across three global sites in the U.S., U.K.and Asia, we are ready to partner with you to help you achieve your next milestone.
Labcorp's Molecular Bioanalytical Services has deep experience in the three main and ancillary types of assays used in the development of cell and gene therapies throughout the drug development life cycle, including biodistribution, vector shedding and vector copy number.
| Biodistribution | Vector Shedding | Vector Copy Number (VCN) | |
|---|---|---|---|
| Definition | Assessment of the distribution of ATMP/GTMP material within a specific in vivo model | Release of virus-based gene therapy product from the animal or patient | VCN is sometimes referred to as cellular kinetics and involves monitoring the persistence and expansion of adoptive cell therapies Sometimes used by sponsors to talk about biodistribution + vector shedding studies as same assay can be used to measure both |
| Nucleic acid measured | DNA – presence of viral vector within an organ/body fluid/excreta RNA – confirm if infection, and subsequent transcription, have occurred or look at up/downregulation of gene expression after dosing | DNA/RNA dependent on viral vector used – look at biofluids, over time, for clearance of viral vector DNA – AVV DNA – any DNA modified viral vector (e.g., Ad-5) RNA – Lentivirus RNA – Lentivirus and retrovirus | DNA/RNA dependent on viral vector used – How much of the transgene or plasmid can we detect in sample cells? DNA – AAV RNA – lentivirus |
| Time points | Animal model dependent but typically 1 week, 1 and 3 months | Usually baseline, then multiple up to a point where levels are no longer detected where 2-3 consecutive negatives are reported | During dosing and long-term follow-up post-dosing |
| Material tested | Various tissues, depending on target organ; may incorporate vector shedding alongside so it could include urine, blood, feces | Animal or patient excreta such as urine, feces, semen, tears, saliva, blood and/or components, CSF | Usually the pre-dose or post-dose material, so usually small sample numbers |
| Assay | qPCR or RT-qPCR as sample numbers are usually large and these assays have a larger dynamic range to cover values from all sample types; see values in range of 10e0 to 10e10+ (dd-PCR or ddRT-qPCR could also be used, but it is very expensive and has less throughput and smaller dynamic range for this study type) | qPCR or RT-qPCR as sample numbers are usually large and these assays have a larger dynamic range to cover values from all sample types; see values in range of 10e0 to 10e10 (dd-PCR or ddRT-qPCR could also be used, but it is very expensive for this study) | ddPCR is a better fit due to the potential for small quantities of the target molecular in complex matrices (and also better precision needed); it’s not linked to the number of samples (although it takes longer and will be more expensive) Recommended for clinical assessments not preclinical studies |
| Platform | QIAsymphony®/KingFisherTM for extraction QuantStudioTM Flex 6 and 7 for PCR | QIAsymphony/KingFisher for extraction QuantStudio Flex 6 and 7 for PCR | QIAsymphony/KingFisher for extraction and Bio-Rad® QX200 for PCR |
| Site | HGT + GRN | HGT + GRN | HGT + GRN |
Table 1: Labcorp Molecular Bioanalytical Services assay list
Labcorp's Molecular Bioanalytical Services also has deep expertise in the four main molecular assay platforms used for the development of cell and gene therapies throughout the drug development life cycle, including qPCR, RT-qPCR, ddPCR and ddRT-PCR.
| qPCR | RT-qPCR | ddPCR | ddRT-PCR | |
|---|---|---|---|---|
| Oligonucleotide | DNA | RNA | DNA | RNA |
| Instrumentation | QuantStudio Flex 6 and 7 | QuantStudio Flex 6 and 7 | Bio-Rad QX200 | Bio-Rad QX200 |
| Well format | 96- or 384-well format | 96- or 384-well format | 96-well format | 96-well format |
| Quantitation | Absolute | Absolute Relative | Absolute | Absolute Relative |
| Dyes | FAM. VIC, JOE, TAMRA, ROX, MGB SYBR Green | FAM. VIC, JOE, TAMRA, ROX, MGB SYBR Green | NA | NA |
| Stage | Preclinical Clinical | Preclinical Clinical | Preclinical Clinical | Preclinical Clinical |
| Applications | Biodistribution Viral shedding of DNA target vcn | Biodistribution Viral shedding of RNA target VCN Up- and downregulation of target gene | Biodistribution Viral shedding of DNA Target VCN | Biodistribution Viral shedding of RNA target VCN Up- and downregulation of target gene |
Table 2: Labcorp Molecular Bioanalytical Services platform list
From preclinical to clinical development needs, Labcorp's Molecular Bioanalytical Services has a range of assay design and testing solutions to help you along the drug development continuum.
| Preclinical | Clinical | |
|---|---|---|
| Types of studies run | Biodistribution
Vector shedding
| Vector shedding
VCN
|
| Automated/manual extractions (HGT + GRN) | DNA extractions – automated extraction RNA extractions – automated extractions | DNA extractions – automated extraction RNA extractions – automated extractions |
| Nucleic acid concentration determination (HGT + GRN) | UV spec – used for biodistribution and vector shedding concentration determination so we can test 1 µg DNA where possible and to assess quality | UV spec – used for vector shedding as for preclinical PicoGreen/RiboGreen nucleic acid determination – used for VCN in order to provide a more accurate copy number quantity |
| Plasmid linearization /digests (HGT + GRN) | Used to cut plasmids as linear plasmid provides more accurate quantitation standard | Used to cut plasmids as linear plasmid provides more accurate quantitation standard |
| PCR assay design (HGT + GRN) | Required for all methods if there is no sponsor-supplied method | Required for all methods if there is no sponsor-supplied method |
| Standard PCR and agarose gel (HGT + GRN) | Only used as required during development to look at DNA/RNA quality alongside UV spec | Only used as required during development to look at DNA/RNA quality alongside UV spec |
| PCR assays (HGT + GRN) | Absolute and relative quantification as detailed previously | Absolute and relative quantification as detailed previously |
| Automated liquid handling (HGT) | To add additional capacity and precision as required | To add additional capacity and precision as required |
Table 3: Labcorp Molecular Bioanalytical Services product lines
