Work Item
ASTM WK76823

New Guide for Evaluation of Immunostimulatory Properties of Nucleic Acid Nanoparticles (NANPs)

1. Scope

This standard will provide guidance on the synthesis and the measurement approaches . The purpose of this document is to provide a guide for applying several standard practices to understand immunostimulatory properties of nucleic acid nanoparticles (NANPs). Expectations and recommendations for NANPs synthesis and characterization by native-PAGE are discussed. Expectations and recommendations for NANPs characterization in the cytokine secretion assay are discussed This guide is intended to serve as an example for manufacturers, producers, analysts, and others with an interest in NANPs. This Standard Guide is not intended to be a comprehensive overview of all possible characterization methods and results. This guide does not include all sample preparation procedures for all possible materials and applications. The user must validate the appropriateness for their particular application. Intended users - industry, academic investigators planning on translating their NANPs to clinical applications, and regulatory agencies


NANPs, nanoparticles, cytokine storm


Immunological properties of DNA and RNA cubes synthesized according to Standard Practice for the preparation of NANPs and their characterization by native-PAGE are well characterized. Therefore, these materials are recommended to be used as benchmark reference materials for analysis according to the Standard Practice for the Preparation and Analysis of Culture Supernatants for the Presence of Cytokine Biomarkers by NANPs in Human Peripheral Blood Mononuclear Cells (PBMCs). When free from contamination with endotoxins and used without a delivery carrier, NANPs are immunoquiescent. After complexation with a carrier that ensures the delivery of NANPs into PBMCs, NANPs acquire immunostimulatory properties. Immunostimulation of intracellularly-delivered NANPs is determined by their physicochemical properties, such as composition (e.g., DNA vs. RNA), dimensionality (e.g., 3D vs. 2D vs. fibrous), and size (e.g., molecular weight of NANPs and their hydrodynamic radii) . The platform for NANPs delivery into the cells, which has historically been a hurdle for therapeutic nucleic acids, is an additional means to tailoring NANP immunorecognition. Identifying and understanding NANPs immunorecognition is important for both the safety and efficacy of NANPs-based therapeutics. NANPs capabilities of inducing cytokines along with the knowledge of the regulation of these properties via a delivery carrier provide important information for NANPs use in vaccines and immunotherapies. Intended users.

The title and scope are in draft form and are under development within this ASTM Committee.


Developed by Subcommittee: E56.03

Committee: E56

Staff Manager: Frank McConnell

Work Item Status

Date Initiated: 05-07-2021

Technical Contact: Marina Dobrovolskaia

Item: 003

Ballot: E56.03 (21-01)

Status: Negative Votes Need Resolution