ASTM_E_2805_-_11.pdf

Designation:E280511Standard Practice forMeasurement of the Biological Activity of Ricin1This standard is issued under the fixed designation E2805;the number immediately following the designation indicates the year oforiginal adoption or,in the case of revision,the year of last revision.A number in parentheses indicates the year of last reapproval.Asuperscript epsilon()indicates an editorial change since the last revision or reapproval.INTRODUCTIONRicin is a member of the protein toxins that cause their physiological effect by inactivation ofribosomes.Ricin is a member of the class 2 ribosome inactivating proteins(1).2Other members of thisclass of toxins include the proteins abrin and Shiga toxin.Ricin consists of two chains,the A-chain that is responsible for the N-glycosidase enzymaticactivity and the B-chain that is needed for cell binding and intra-cellular processing.Ricin is aheterogeneous protein with molecular weights ranging from approximately 62 to 64 kilodaltons(kDa)(2).Both chains are glycosylated and of similar size(approximately 32 kDa).There are several genesencoding putative ricin and ricin-like proteins in the genome of R.communis(3)resulting indifferences in the amino acid sequence of the subunits.The differences in amino acid sequence andglycosylation both contribute to the heterogeneity of ricin.1.Scope1.1 This guide is intended for the manufacturers and usersof ricin reference material.Ricin reference materials arewell-characterized materials that can be used to test detectiondevices and calibrate laboratory measurements.It is anticipatedthat ricin reference materials will be characterized by bio-chemical methods in addition to the measurement of biologicalactivity.1.2 This practice details the measurement of ricin biologicalactivity using a cell-free translation(CFT)assay(4).1.3 The CFT assay has been developed for use in anybiotechnology laboratory where determination or confirmationof ricin biological activity is required.1.4 The CFT assay has been validated by the U.S.ArmyMedical Research Institute of Infectious Diseases(USAM-RIID)VP-016 Validation of Cell-Free Translation Assay forthe Detection of Ricin Toxin Biological Activities in compli-ance(5)with Good Laboratory Practices(GLP)Regulations ofthe Food and Drug Administration(21 CFR Part 58).Strictadherence to the protocol is necessary for validity of the testresults.1.5 Appendix X1 and Appendix X2 also provide guidancefor the measurement of the biological activity of ricin usingcell-based assays and the use of synthetic enzyme substrates.1.6 Ricin is a category 2 select agent and acquisition of thericin standard must adhere to the Center for Disease Control(CDC)regulations.Ricin is listed on the select agent list(42CFR Part 72).3The possession,transfer,and use of ricin arerestricted under the Public Health Security Preparedness Act(CRS Report RL31263 Public Health Security and Bioterror-ism Preparedness and Response Act(P.L.107-188):Provisionand Changes to Preexisting law).Access to stores of ricin islimited(USA Patriot Act,P.L.107-56).Ricin is also aprohibited substance under the Biological Weapons Conven-tion and the Chemical Weapons Convention(CRS ReportRL31559 Proliferation Control Regimes:Background andStatus).1.7 The values stated in SI units are to be regarded asstandard.No other units of measurement are included in thisstandard.1.8 This standard does not purport to address all of thesafety concerns,if any,associated with its use.It is theresponsibility of the user of this standard to establish appro-priate safety and health practices and determine the applica-bility of regulatory limitations prior to use.Ricin is anextremely dangerous toxin.See Section 9 for specific hazardsinformation.1This practice is under the jurisdiction of ASTM Committee E54 on HomelandSecurity Applications and is the direct responsibility of Subcommittee E54.01 onCBRNE Sensors and Detectors.Current edition approved Jan.1,2011.Published March 2011.DOI:10.1520/E2805-11.2The boldface numbers in parenthesis refer to the list of references at the end ofthis standard.3Available at http:/www.bt.cdc.gov/Agent/agentlist.asp.Copyright ASTM International,100 Barr Harbor Drive,PO Box C700,West Conshohocken,PA 19428-2959.United States1 2.Referenced Documents2.1 ASTM Standards:4F2149 Test Method for Automated Analyses of CellstheElectrical Sensing Zone Method of Enumerating andSizing Single Cell Suspensions2.2 Code of Federal Regulations:21 CFR Part 58 Good laboratory practice for nonclinicallaboratory studies542 CFR Part 72 Interstate shipment of etiologic agents62.3 ANSI/ATCC Standard:7ASN-0001-2009 Standardization of In-Vitro Assays to De-termine Anthrax Toxin Activities3.Terminology3.1 Abbreviations:3.1.1 CFTcell free translation.3.1.2 CPScounts per second,units of luminescence instru-ment.3.1.3 IC50concentration of ricin that produces inhibitionof 50%of the activity in an assay.3.1.4 kDamolecular mass in kilo Dalton units.3.1.5 PBSphosphate buffered saline.