ASTM F3515-21

This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
Designation: F3515 − 21
Standard Guide for
Characterization and Testing of Porcine Fibrinogen as a
Starting Material for Use in Biomedical and Tissue-
Engineered Medical Product Applications
This standard is issued under the fixed designation F3515; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope 1.7 This international standard was developed in accor-
dance with internationally recognized principles on standard-
1.1 This guide covers the evaluation of porcine fibrinogen
ization established in the Decision on Principles for the
suitable for use in biomedical or pharmaceutical applications
Development of International Standards, Guides and Recom-
including, but not limited to, tissue-engineered medical prod-
mendations issued by the World Trade Organization Technical
ucts (TEMPs).
Barriers to Trade (TBT) Committee.
1.2 This guide addresses key parameters relevant for
functionality, characterization, and purity of porcine fibrino-
2. Referenced Documents
gen.
2.1 ASTM Standards:
1.3 As with any material, some characteristics of porcine
E1298 Guide for Determination of Purity, Impurities, and
fibrinogen may be altered by processing techniques, such as
Contaminants in Biological Drug Products (Withdrawn
electrospinning (1) and sterilization, required for the produc-
2014)
tion of a specific formulation or device. Therefore, properties
F981 Practice for Assessment of Compatibility of Biomate-
of fabricated forms of this protein should be evaluated using
rials for Surgical Implants with Respect to Effect of
test methods that are appropriate to ensure safety and efficacy
Materials on Muscle and Insertion into Bone
and are not addressed in this guide.
F1983 Practice forAssessment of Selected Tissue Effects of
1.4 The primary focus of this document is fibrinogen Absorbable Biomaterials for Implant Applications
F2212 Guide for Characterization of Type I Collagen as
derived from porcine blood, which is similar to human fibrino-
gen. The biggest advantage that pigs have over other species Starting Material for Surgical Implants and Substrates for
(such as cattle, sheep, goats, elk, and deer) is that they are less Tissue Engineered Medical Products (TEMPs)
likely to transmit transmissible spongiform encephalitis (TSE) F3163 Guide for Classification of Cellular and/or Tissue-
(ISO 22442-1 Annex D; WHO Guidelines, 2003; WHO Based Products (CTPs) for Skin Wounds
Guidelines,2006;WHOGuidelines,2010).Thedocumentmay
2.2 ISO Standards:
also discuss fibrinogen from other sources when useful infor-
ISO10993-1 Biologicalevaluationofmedicaldevices—Part
mation is available. Fibrin is also discussed in some sections.
1: Evaluation and testing with a risk management process
1.5 Units—The values stated in SI units are to be regarded ISO 11137-1:2006 Sterilization of health care products—
Radiation—Part 1: Requirements for validation and rou-
as the standard. No other units of measurement are included in
this standard. tine control of a sterilization process for medical devices
ISO 11737-1:2006 Sterilization of health care products—
1.6 This standard does not purport to address all of the
Microbiological methods—Part 1: Determination of a
safety concerns, if any, associated with its use. It is the
population of microorganisms on products
responsibility of the user of this standard to establish appro-
ISO 11737-2:1998 Sterilization of health care products—
priate safety, health, and environmental practices and deter-
mine the applicability of regulatory limitations prior to use.
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
This guide is under the jurisdiction of ASTM Committee F04 on Medical and contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
Surgical Materials and Devices and is the direct responsibility of Subcommittee Standards volume information, refer to the standard’s Document Summary page on
F04.42 on Biomaterials and Biomolecules for TEMPs. the ASTM website.
Current edition approved Aug. 1, 2021. Published August 2021. DOI: 10.1520/ The last approved version of this historical standard is referenced on
F3515-21. www.astm.org.
2 5
The boldface numbers in parentheses refer to a list of references at the end of Available fromAmerican National Standards Institute (ANSI), 25 W. 43rd St.,
this standard. 4th Floor, New York, NY 10036, http://www.ansi.org.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
F3515 − 21
Microbiological methods—Part 2: Tests of sterility per- ISO 22442-2 Medical devices utilizing animal tissues and
formed in the definition, validation and maintenance of a their derivatives—Part 2: Controls on sourcing, collection
and handling
sterilization process
ISO 13485 Medical devices—Quality management ISO 22442-3 Medical devices utilizing animal tissues and
their derivatives—Part 3: Validation of the elimination
systems—Requirements for regulatory purposes
and/or inactivation of viruses and transmissible spongi-
ISO 14160:2020 Sterilization of health care products—
form encephalopathy (TSE) agents
Liquid chemical sterilizing agents for single-use medical
devices utilizing animal tissues and their derivatives—
2.3 Other Documents:
Requirements for characterization, development, valida-
AAMI TIR 19:1998 Guidance for ANSI/AAMI/ISO 10993-
tion and routine control of a sterilization process for
7:1995, Biological evaluation of medical devices—Part 7:
medical devices
Ethylene oxide sterilization residuals
ISO 14644-1 Cleanrooms and associated controlled 21 CFR 211 Current Good Manufacturing Practice for Fin-
environments—Part 1: Classification of air cleanliness by
ished Pharmaceuticals
particle concentration 21 CFR 820 Quality System Regulation
European Commission, OJ L 2011/C 73/01 Note for guid-
ISO 14644-2 Cleanrooms and associated controlled
environments—Part 2: Monitoring to provide evidence of ance on minimizing the risk of transmitting animal spon-
giform encephalopathy agents via human and veterinary
cleanroom performance related to air cleanliness by par-
medicinal products (EMA/410/01 rev.3)
ticle concentration
European Pharmacopoeia 9.0, 2017 Fibrin Sealant Kit
ISO 14644-3 Cleanrooms and associated controlled
European Pharmacopoeia 9.0, 2017, Chapter 2.2.3 Potentio-
environments—Part 3: Test methods
metric Determination of pH
ISO 14644-4 Cleanrooms and associated controlled
European Pharmacopoeia 9.0, 2017, Chapter 2.2.40 Near-
environments—Part 4: Design, construction and start-up
Infrared Spectroscopy
ISO 14644-5 Cleanrooms and associated controlled
European Pharmacopoeia 9.0, 2017, Chapter 2.4.8 Heavy
environments—Part 5: Operations
Metals
ISO 14644-7 Cleanrooms and associated controlled
European Pharmacopoeia 9.0, 2017, Chapter 2.5.9 Determi-
environments—Part 7: Separative devices (clean air
nation of Nitrogen by Sulphuric Acid Digestion
hoods, gloveboxes, isolators and mini-environments)
European Pharmacopoeia 9.0, 2017, Chapter 2.5.12 Water:
ISO 14644-8 Cleanrooms and associated controlled
Semi-micro Determination
environments—Part 8: Classification of air cleanliness by
European Pharmacopoeia 9.0, 2017, Chapter 2.5.32 Loss on
chemical concentration (ACC)
Drying
ISO 14644-9 Cleanrooms and associated controlled
European Pharmacopoeia 9.0, 2017, Chapter 2.6.1 Sterility
environments—Part 9: Cleaning of surfaces to achieve
European Pharmacopoeia 9.0, 2017, Chapter 2.6.12 Total
defined levels of cleanliness in terms of particle and
Viable Aerobic Count
chemical classifications
European Pharmacopoeia 9.0, 2017, Chapter 2.6.13 Test for
ISO 14644-10 Cleanrooms and associated controlled
Specified Microorganisms
environments—Part 10: Classification of surface cleanli-
European Pharmacopoeia 9.0, 2017, Chapter 2.6.14 Bacte-
ness by chemical concentration
rial Endotoxins
ISO 14644-13 Cleanrooms and associated controlled
European Pharmacopoeia 10.0, Supplement 10.3, 2020,
environments—Part 13: Cleaning of surfaces to achieve
Chapter 2.6.32 Test for Bacterial Endotoxins Using Re-
defined levels of cleanliness in terms of particle and
combinant Factor C
chemical classifications
European Pharmacopoeia 9.0, 2017, Chapter 5.1.6 Alterna-
ISO 14644-14 Cleanrooms and associated controlled
tive Methods for Control of Microbiological Quality
environments—Part 14: Assessment of suitability for use
FDA Guidance Document, 1991 Shelf Life of Medical
of equipment by airborne particle concentration
Devices
ISO 14644-15 Cleanrooms and associated controlled
FDA Guidance Document, 2008 Guidance for Industry:
environments—Part 15: Assessment of suitability for use
Container and Closure System IntegrityTesting in Lieu of
of equipment and materials by airborne chemical concen-
tration
ISO 14698-1 Cleanrooms and associated controlled
Available from Association for the Advancement of Medical Instrumentation
environments—Biocontamination control—Part 1: Gen-
(AAMI), 4301 N. Fairfax Dr., Suite 301, Arlington, VA 22203-1633, http://
eral principles and methods www.aami.org.
Available from U.S. Government Printing Office, Superintendent of
ISO 14698-2 Cleanrooms and associated controlled
Documents, 732 N. Capitol St., NW, Washington, DC 20401-0001, http://
environments—Biocontamination control—Part 2: Evalu-
www.access.gpo.gov.
ation and interpretation of biocontamination data
Available from the European Medicines Agency (EMA), Domenico Scarlatti-
laan 6 1083 HS Amsterdam, The Netherlands, https://www.ema.europa.eu.
ISO 14971 Medical devices—Application of risk manage-
Available from the EDQM Council of Europe, 7 allée Kastner, CS 30026,
ment to medical devices
F-67081 Strasbourg, France, https://pharmeuropa.edqm.eu.
ISO 22442-1 Medical devices utilizing animal tissues and
Available from U.S. Food and Drug Administration (FDA), 10903 New
their derivatives—Part 1:Application of risk management Hampshire Ave., Silver Spring, MD 20993, http://www.fda.gov.
F3515 − 21
Sterility Testing as a Component of the Stability Protocol ligaments, fascia, skin, fibrous tissues, fat, and synovial mem-
for Sterile Products branes (which are connective tissue), and muscles, nerves, and
FDA Guidance Document, 2012 Guidance for Industry: blood vessels (which are not connective tissue).
Pyrogen and Endotoxins Testing: Questions and An-
3.1.6 tissue-inducing biomaterials, n—lifeless biomaterials
swers
capable of inducing tissue regeneration in vivo without addi-
ICH Q1A(R2), 2003 Stability Testing of New Drug Sub-
tion of growth factors or living cells (3).
stances and Products
3.1.7 tissue regeneration, n—ahealingprocessinwhichlost
ICH Q7, 2000 Good Manufacturing Practice for Active
or damaged tissue is placed by migration, differentiation,
Pharmaceutical Ingredients
proliferation, and patterning of cells that deposit extracellular
USP <61> Microbial Limit Tests
matrix with normal architecture, function, and topological
USP<62> Microbiological Examination of Nonsterile Prod-
appearance. F3163
ucts: Tests for Specified Microorganisms
USP <71> Sterility Tests
4. Summary of Guide
USP <85> Bacterial Endotoxins Test
4.1 Fibrinogen, a soluble and complex plasma glycoprotein
USP <231> Heavy Metals
in vertebrate blood, is a hexamer, containing two sets of three
USP <731> Loss on Drying
different chains (α, β, and γ), having a total molecular mass of
USP <791> pH
340 000 g/mol. The use of fibrinogen derived from porcine
USP <1211> Sterilization and Sterility Assurance of Com-
blood in surgery started in approximately 2000 when fibrin gel
pendial Articles
formed by mixing fibrinogen and thrombin could effectively
World Health Organization, 2003 WHO Guidelines on
stop bleeding during surgery. More recently, fibrinogen, as a
Transmissible Spongiform Encephalopathies in Relation
raw material, has been blended with a degradable polymer and
to Biological and Pharmaceutical Products
prepared into a micro- to nano-scale three-dimensional net-
World Health Organization, 2006 WHO Guidelines on Tis-
work structure by electrospinning (4, 5) for use in the regen-
sue Infectivity Distribution in Transmissible Spongiform
eration of soft tissue as a tissue-inducing biomaterial (4, 6).
Encephalopathies
The aim of this guide is to identify key parameters relevant for
World Health Organization, 2010 WHO Tables on Tissue
the functionality and characterization of fibrinogen during
Infectivity Distribution in Transmissible Spongiform 188
developmentofnewproductscontainingporcinefibrinogenfor
Encephalopathies
biomedical applications.
3. Terminology
5. Significance and Use
3.1 Definitions of Terms Specific to This Standard:
5.1 The purpose of this guide is to provide guidance on
3.1.1 electrospinning, n—a fiber manufacturing process to
characterization of the properties of porcine fibrinogen as a
produce ultrafine fibres (fiber diameter range of nanometers to
starting material for surgical implants and as a matrix for
micrometers) by charging and ejecting a polymer melt or
tissue-engineered medical products (TEMPs). This guide con-
polymer solution through a spinneret under a high-voltage
tains a set of physical and chemical parameters directly related
electric field and to solidify or coagulate it to form a filament
to the function of porcine fibrinogen.This guide can be used to
(2).
help select and characterize appropriate fibrinogen starting
3.1.2 factor XIII, n—a factor that polymerizes fibrin mono-
materials for specific purposes. Not all tests or parameters are
mers so that they become stable and insoluble in urea, thus
suitable for all uses of fibrinogen.
enabling fibrin to form a firm blood clot (also known as
fibrin-stabilizing factor (FSF)). 5.2 Fibrinogen described in this guide may be used in
various types of medical products including, but not limited to,
3.1.3 fibrin, n—the insoluble protein formed from fibrino-
implants, tissue-engineered medical products (TEMPs), and
gen by the proteolytic action of thrombin during normal
cell, drug, or DNA delivery vectors. The recommendations in
clotting of blood; it forms the essential portion of the blood
this guide shall not be construed to guarantee the successful
clot.
clinical application of any tissue-engineered medical product.
3.1.4 fibrinogen, n—a plasma protein with an approximate
5.3 In determining whether fibrinogen meets the require-
molecular mass of 340 000 g/mol, composed of three subunits
ments for use in a TEMP, the relevant regulatory authorities or
(α,β,γ)encodedbyseparategenes,whichisconvertedtofibrin
other appropriate guidelines relating to the production,
throughtheactionofthrombinandisakeyproteinintheblood
regulation, and approval of TEMP products shall be taken into
clotting cascade.
account (Guide E1298, Practice F981, Practice F1983).
3.1.5 soft tissue, n—the tissues that connect, support, or
surround other structures and organs of the body, not being
6. Physical and Chemical Characterization Methods
hard tissue such as bone. Soft tissue includes tendons,
6.1 Characteristics of Fibrinogen:
6.1.1 Solubility (European Pharmacopoeia 9.0, Fibrin Seal-
Available from U.S. Pharmacopeial Convention (USP), 12601 Twinbrook
ant Kit)—Fibrinogen should be dissolved at an appropriate
Pkwy., Rockville, MD 20852-1790, http://www.usp.org.
temperature (often 37 °C) in an appropriate medium (such as
Available from World Health Organization (WHO), Avenue Appia 20 1211
Geneva, Switzerland, https://www.who.int. an aqueous buffer solution) for an appropriate amount of time
F3515 − 21
(often 20 min), and the clarity of the solution should be 6.1.7.3 Nicotinamide adenine dinucleotide hydrogen
reported by visual assessment. Typically, soluble fibrinogen (NADH) reagent, containing glutamate dehydrogenase,
α-ketoglutarate, and NADH in a suitable buffer solution.
solutions are
...