ASTM_F_2065_-_00_2010.pdf

Designation:F206500(Reapproved 2010)Standard Practice forTesting for Alternative Pathway Complement Activation inSerum by Solid Materials1This standard is issued under the fixed designation F2065;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.1.Scope1.1 This practice provides a protocol for rapid,in vitroscreening for alternative pathway complement activating prop-erties of solid materials used in the fabrication of medicaldevices that will contact blood.1.2 This practice is intended to evaluate the acute in vitroalternative pathway complement activating properties of solidmaterials intended for use in contact with blood.For thispractice,“serum”is synonymous with“complement.”1.3 This practice consists of two procedural parts.Proce-dureAdescribes exposure of solid materials to a standard lot ofC4-deficient guinea pig serum C4(-)GPS,using 0.1-mLserum per 13 100-mm disposable glass test tubes.Sepharose2CL-4B is used as an example of test materials.Procedure Bdescribes assaying the exposed serum for significant functionalalternative pathway complement depletion as compared tocontrol samples.The endpoint in procedure B is lysis of rabbitRBC in buffer containing EGTA and excess Mg+.1.4 This practice does not address function,elaboration,ordepletion of individual complement components except asoptional additional confirmatory information that can be ac-quired using human serum as the complement source.Thispractice does not address the use of plasma as a source ofcomplement.1.5 This practice is one of several developed for theassessment of the biocompatibility of materials.Practice F748may provide guidance for the selection of appropriate methodsfor testing materials for other aspects of biocompatibility.Practice F1984 provides guidance for testing solid materials forwhole complement activation in human serum,but does notdiscriminate between the classical or alternative pathway ofactivation.1.6 The values stated in SI units are to be regarded asstandard.No other units of measurement are included in thisstandard.2.Referenced Documents2.1 ASTM Standards:3F748 Practice for Selecting Generic Biological Test Methodsfor Materials and DevicesF1984 Practice for Testing for Whole Complement Activa-tion in Serum by Solid Materials2.2 Other Document:ISO 10993-4:Biological Evaluation of Medical Devices.Part 4:Selection of Tests for Interactions with Blood43.Terminology3.1 Definitions of Terms Specific to This Standard:3.1.1 waterdistilled,endotoxin-free.3.2 Abbreviations:3.2.1 Abantibody(hemolysin)3.2.2 BBSbarbital buffered saline3.2.3 BBS-Gbarbital buffered saline gelatin3.2.4 BBS-G-EGTA/Mg(Mg Buffer)barbital buffered sa-line gelatin EGTA Mg+3.2.5 BBS-GM(Ca Buffer)barbital buffered saline gela-tin metals3.2.6 Ccomplement3.2.7 C4(-)GPSC4-deficient guinea pig serum serumfrom guinea pigs genetically incapable of producing C4,thefourth component of complement3.2.8 EDTAethylenediaminetetraacetic acid,disodiumsalt:dihydrate3.2.9 EGTAethylene glyco-bis(b-aminoethyl ether)-N,N,N,N-tetraacetic acid,tetrasodium salt1This practice is under the jurisdiction ofASTM Committee F04 on Medical andSurgical Materials and Devices and is the direct responsibility of SubcommitteeF04.16 on Biocompatibility Test Methods.Current edition approved Sept.1,2010.Published November 2010.Originallyapproved in 2000.Last previous edition approved in 2006 as F2065 00(2006).DOI:10.1520/F2065-00R10.2Sepharose is a registered trademark of Pharmacia,Inc.(now GE Healthcare),Amersham Place,Little Chalfont,Buckinghamshire HP7 9NA,U.K.3For referenced ASTM standards,visit the ASTM website,www.astm.org,orcontact ASTM Customer Service at serviceastm.org.For Annual Book of ASTMStandards volume information,refer to the standards Document Summary page onthe ASTM website.4Available from American National Standards Institute(ANSI),25 W.43rd St.,4th Floor,New York,NY 10036.Copyright ASTM International,100 Barr Harbor Drive,PO Box C700,West Conshohocken,PA 19428-2959.United States1 3.2.10 HAGGheat aggregated gamma globulin3.2.11 HShuman serum3.2.12 Icontrol tube with serum but no material,kept onice.3.2.13 Mtube containing serum plus a test material3.2.14 NMtube containing serum but no material3.2.15 PVDFpolyvinylidene fluoride3.2.16 RBCred blood cell(s)4.Summary of Practice4.1 Solid material specimens are exposed to a standard lotof C4(-)GPS complement under defined conditions,in parallelto appropriate controls(Procedure A).If the alternativecomplement pathway is activated by the material,complementcomponents will be depleted from the serum.Exposed serum isthen tested for remaining functional complement activity,bydetermining complement mediated lysis of rabbit RBC inbuffer containing EGTA and excess Mg+(Procedure B).5.Significance and