Analytical Development

Based on Next Generation Sequencing (NGS) technology, IDT Biologika uses its own developed bioinformatics analysis pipeline to conduct Adventitious Agents Testing (AVT). NGS is much faster than conventional animal testing, classical qPCR-approach, and in-vitro tests.

For the detection of DNA impurities from the species Gallus Gallus (e.g. by using DF-1 manufacturing cell lines), IDT Biologika developed the chicken residual host cell DNA (hcDNA) assay. It is a quantitative PCR (qPCR)-based assay with high sensitivity and specificity. The results are generated within a few hours by a streamlined workflow consisting of DNA extraction and subsequent qPCR analysis.

IDT Biologika developed and validated a qPCR assay for fast determination of a broad range of mycoplasma, including non-cultivable mycoplasma and those which cannot be detected with the classical indicator method. With this assay, a significant reduction in the testing timeline is achieved.

IDT Biologika offers a fast mycobacteria test using qPCR. It provides results after just three days, whereas the classical microbiological method takes about of eight weeks. With this new test method, a broad range of mycobacteria species are detectable within a sample volume of 10 ml.

Digital droplet PCR ddPCR is a highly sensitive technology for the quantification of DNA/RNA, without the need for a DNA/RNA standard. Therefore, the development of such an assay is faster than using a classical qPCR.

At IDT, ddPCR is used for a broad range of different analytics. This includes quantification of virus material, examination and quantification of viral/bacterial contaminations, and characterisation of control material. In addition, ddPCR is used for most of the genome titer determination assays.

For cell based potency determination and characterization of viral vaccines, IDT Biologika offers fluorescence-based flow cytometry as a more rapid and more objective analysis than conventional plaque forming cells and dilution techniques.

For fingerprint analytics of protein profiles in viral vaccines on different process steps, IDT Biologika offers Capillary Electrophoresis analytics (SDS-PAGE mode). Here, the specific protein profile of different process steps can be visualized for characterization of the downstream process, and also specific protein identification and quantitation is possible.

To identify and quantify proteins in a highly specific manner, for process characterization or as impurity analysis, IDT Biologika offers high-resolution mass spectrometry (Q-TOF) and Triple-Quad-LC-MS/MS. It may be utilized for viral protein expression studies on viral vaccines or as highly-sensitive impurity quantitation for known peptide or protein impurities.

For endotoxin testing, all classical methods are used – kinetic turbidimetric and kinetic chromogenic, as well as gel clot method. For rapid endotoxin testing of IPCs, the cartridge method using the Endosafe® nexgen-PTS is available. Endotoxin testing is performed on IPC and EPC samples. Furthermore, incoming goods including primary packaging material (stoppers and vials) are tested for endotoxins. Low endotoxin recovery studies may be performed if required.