Hans Baumeister, Managing Director at FyoniBio discussed how clinical and bioanalytical techniques can be used for quantification and immunogenicity assessments of oligonucleotides.
FyoniBio, a solution provider based in Germany provides bioanalytical support clinical trials, assay development and process analytics. The focus of the company spans various modalities including antibodies, glycoproteins and oligonucleotides. Upon expanding into oligonucleotide research, the company focuses on siRNA and therapeutic oligonucleotides.
Baumeister further elaborated on his team’s expertise in developing customized assays for pharmacokinetics (PK), pharmacodynamics (PD), and immunogenicity, validated under GCLP standards. Key challenges in PK assays include matrix effects, sensitivity, and metabolite specificity. Therefore, establishing robust, high-throughput assays with minimal sample processing is essential.
Baumeister compared techniques such as LC-MS, Stem-Loop qPCR, and hybridization-enhanced electrochemiluminescence (hECLIA). The latter, preferred for its speed, transferability, and high sensitivity, is particularly effective for detecting oligonucleotides in biological samples with picomolar precision. However, LC-MS remains essential for analysing truncated oligonucleotides and metabolites.
Baumeister outlined immunogenicity testing, focusing on the detection of anti-drug antibodies (ADAs). Despite therapeutic oligonucleotides being less known for immunogenicity than RNA, ADAs have been reported in cases like mipomersen and patisiran.
Baumeister stated: “For Mipomersen, 72% of patients responded with ADA to this drug.” He detailed the stages of ADA detection: risk assessment, assay development, and analysis during clinical trials.
Challenges include designing specific assays for oligonucleotides and determining accurate cut points amidst pre-dose signal variation. The team addressed these by using patient-specific cut points and advanced sandwich formats, ensuring sensitive and precise assays with high drug tolerance.
Baumeister concluded with a summary of FyoniBio’s achievements in oligonucleotide quantification and ADA analysis, noting robust, high-throughput methodologies and advancements in assay sensitivity and drug tolerance.
