Large molecule biotherapeutics are primarily expressed and produced in mammalian host cells then purified from complex mixtures containing growth media and host cell material. Identifying, monitoring, and eliminating host cell proteins (HCP) and host cell DNA (hcDNA) are crucial to prevent any adverse immune response, toxicity or interference with the product’s biological activity.
While most HCPs can be eliminated effectively, monitoring especially some low-abundance HCPs can be difficult and challenging with various analytical methods such as the widely used ELISA technique and LC-MS. The lack of well annotated HCP sequence databases also impacts identification.
Similar challenges are encountered with the removal of hcDNA. Chromatin (genomic DNA and histone proteins) interferes with purification steps by binding to protein A resin and elutes with the therapeutic product impacting yield.
“We offer a suite of RUO platforms to detect both hcDNA and HCPs for multiple host cell types that are commonly used for the expression of therapeutic proteins and antibodies, or applications in cell and gene therapy (CGT),” said Anis Khimani, PhD, Senior Strategy Leader, Life Sciences at Revvity.
For hcDNA the qPCR-based portfolio, HostDetect
PCR DNA Quant Kits, offer independent genome DNA detection molecular assays for three host cell types – CHO, HEK293, and E.coli. The quick and flexible assays offer high sensitivity, a wide dynamic range that minimizes need for multiple dilutions, and precision that meets ICH guidelines.
HostDetect DNA assay set-up enables flexibility in sample input volume and scalability up to 192 reactions per kit. The speed allows for in-process monitoring on real-time PCR thermal cyclers rather than end-point testing. Built-in internal control, increasingly expected to comply with GMP testing, monitors sample preparation, PCR inhibition, reagent performance, and instrument function.
“For the detection and monitoring of HCPs, we offer no-wash immunoassay kits based on bead and TR-FRET technologies, compatible with CHO and HEK293 host cells,” added Khimani. The AlphaLISA and HTRF protein assays provide low levels of detection, speed and throughput facilitating same day in-process monitoring, a wide dynamic range, and a broad spectrum of antibody assays to detect between 96% – 99.8% coverage of HCPs.
The assays are designed for ease of use and efficiency. They substantially reduce hands-on time and total assay duration, delivering results in less than 3 hours. Streamlining the process while maintaining high sensitivity and reproducibility, the AlphaLISA and HTRF protein assays are tailored to specific host cell types. The platforms are also well-suited for automation and enable precise and robust real-time monitoring of HCPs in a rapid and higher throughput workflow to support purity of the end-product.
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