What is the lower limit of quantification (LLOQ) for your qPCR-based shedding assays for LNP-siRNA?

A: Our GxP-validated qPCR assays are optimized for each specific siRNA sequence and LNP formulation. We consistently achieve LLOQs in the low copies/mL or copies/µg range, tailored to the specific sample matrix and regulatory requirements for your program.

Which sample matrices do you validate for shedding analysis in LNP studies?

A: We validate assays across a comprehensive range of matrices relevant to expected routes of excretion and exposure. Standard matrices include urine, feces, saliva, and blood (plasma/serum). We can also develop and validate methods for nasal swabs, tears, or other matrices as dictated by the route of administration.

How does LNP biodegradability impact the interpretation of shedding data for regulatory submissions?

A: Biodegradability is a primary determinant of the excretion profile. Biodegradable LNPs are expected to exhibit a finite shedding window, demonstrating clearance. Non-biodegradable formulations may show prolonged persistence. Our analysis provides the quantitative data to build a robust safety narrative around clearance kinetics for FDA and EMA review.

Can your assays distinguish between the LNP carrier and the siRNA payload in shed samples?

A: Yes. We employ a multi-assay strategy. A qPCR-based assay quantifies the specific siRNA payload sequence. Concurrently, LC-MS/MS methods can be developed through our strategic partners to quantify specific lipid components of the LNP, providing a comprehensive view of how the construct is cleared.

The selection of lipid nanoparticle (LNP) chemistry directly influences the safety and regulatory trajectory of siRNA therapeutics. Biodegradable ionizable lipids present a significant advantage by mitigating long-term bioaccumulation risks. Verifying their clearance requires highly sensitive, GxP-compliant bioanalytical assays to accurately characterize the excretion profile. This analysis provides a foundational dataset for demonstrating control over the therapeutic construct and satisfying regulatory expectations for environmental risk assessment.

For siRNA therapeutics delivered via LNPs, regulatory bodies require a comprehensive characterization of biodistribution, persistence, and excretion. The objective is to quantify the amount of therapeutic construct shed from the subject, the route of excretion, and the duration of shedding. This data informs the environmental risk assessment (ERA) and is a key component of the pharmacology and toxicology sections of an Investigational New Drug (IND) application. An incomplete or ambiguous shedding dataset can lead to significant delays in clinical progression.

Our approach focuses on generating definitive, submission-ready data that clearly delineates the clearance kinetics of your specific LNP-siRNA formulation. This allows your team to build a robust safety narrative, anticipating and addressing regulatory questions before they are asked.

The fundamental difference in shedding profiles is driven by the LNP’s metabolic fate. The design of novel delivery systems, such as the biodegradable ionizable lipids developed for potent mRNA delivery (PMID: 40060499), aims to ensure the construct is cleared after fulfilling its therapeutic function.

• Biodegradable LNPs: These are engineered with cleavable esters or other labile linkers. They are metabolized and cleared through endogenous pathways, resulting in a transient excretion profile. The analytical challenge is to prove this clearance with assays sensitive enough to detect the construct down to baseline levels.

• Non-biodegradable LNPs: Lacking a mechanism for rapid metabolic breakdown, these lipids can persist longer. While effective for delivery, as demonstrated in early non-viral gene-liposome complexes (PMID: 26149841), their prolonged tissue residence time necessitates a longer-term shedding analysis to define the full clearance profile.

Franklin Biolabs provides the specialized infrastructure and expertise required for these sensitive studies. Our >100,000 sq ft GxP-compliant facility is designed for complex bioanalysis, supporting programs from discovery through IND submission. We have refined our processes to support an accelerated 18-24 month IND timeline without compromising data quality. This operational efficiency is central to our mission.

MAKING LIFE-SAVING RESEARCH AFFORDABLE

Our 100% IND approval success rate since 2019 (with the Franklin Biolabs brand launching in 2024) is a direct result of this rigorous, quality-driven approach. We develop and validate custom qPCR assays to precisely quantify your siRNA payload in diverse biological matrices, providing the clear, quantitative data required by regulators.

All in vivo studies are conducted in full compliance with AAALAC and USDA guidelines. By optimizing sample collection schemes and employing highly sensitive assays, we minimize the number of animals required while maximizing the data obtained from each subject, ensuring both ethical conduct and programmatic efficiency.