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ASTM E2888-12(2019)

(Practice)

Standard Practice for Process for Inactivation of Rodent Retrovirus by pH

Standard Practice for Process for Inactivation of Rodent Retrovirus by pH

SCOPE
1.1 This practice assures 5 log10 inactivation of non-defective C-type retroviruses, which are endogenous to murine hybridoma and CHO cells and are potentially present in the production stream of biopharmaceutical processes that use rodent derived cell culture.  
1.2 The process parameters specified in this practice consistently assure 5 log10 inactivation of murine retrovirus by adjusting the pH of a process solution after initial affinity capture chromatography purification.  
1.3 This practice is applicable to mAb, IgG fusion, or other recombinant proteins produced from rodent cell lines (for example, CHO or murine hybridoma), which do not target retroviral proteins. Additionally, the low pH step is performed on a cell-free intermediate, post initial capture using protein A chromatography.  
1.4 The 5 log10 inactivation of murine retrovirus claimed by using this practice will be utilized in conjunction with other clearance unit operations (for example, chromatography and virus retentive filtration) to assure sufficient total process clearance of murine retroviruses, which will be supportive of early phase regulatory filings.  
1.5 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.6 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.

General Information

Status
Published
Publication Date
31-Jan-2019
ICS
07.100.01 - Microbiology in general
Technical Committee
E55 - Manufacture of Pharmaceutical and Biopharmaceutical Products
Drafting Committee
E55.12 - Process Applications
Current Stage
Ref Project

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ASTM E2888-12(2019) - Standard Practice for Process for Inactivation of Rodent Retrovirus by pHASTM E2888-12(2019) - Standard Practice for Process for Inactivation of Rodent Retrovirus by pH
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ASTM E2888-12(2019) - Standard Practice for Process for Inactivation of Rodent Retrovirus by pH
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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: E2888 − 12 (Reapproved 2019)
Standard Practice for
Process for Inactivation of Rodent Retrovirus by pH
This standard is issued under the fixed designation E2888; 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 quence (usually a human receptor-like protein or protein
fragment) fused to the carboxyl-terminal of the Fc-domain of a
1.1 This practice assures 5 log10 inactivation of non-
human IgG antibody.
defective C-type retroviruses, which are endogenous to murine
2.1.1.1 Discussion—Dimerization occurs by way of the Fc
hybridoma and CHO cells and are potentially present in the
domain.
production stream of biopharmaceutical processes that use
rodent derived cell culture.
2.1.2 immunoglobulin G (IgG), n—an antibody molecule
composed of four peptide chains — two γ heavy chains and
1.2 The process parameters specified in this practice con-
sistently assure 5 log10 inactivation of murine retrovirus by two light chains.
adjusting the pH of a process solution after initial affinity
2.1.2.1 Discussion—Each IgG has two antigen binding
capture chromatography purification.
sites. IgG constitutes 75 % of serum immunoglobulins in
humans. IgG molecules are synthesized and secreted by plasma
1.3 This practice is applicable to mAb, IgG fusion, or other
B cells. There are four IgG subclasses (IgG1, 2, 3, and 4) in
recombinant proteins produced from rodent cell lines (for
humans, named in order of their abundance in serum (IgG1
example, CHO or murine hybridoma), which do not target
being the most abundant). Only human IgG1, IgG2, and IgG4
retroviral proteins. Additionally, the low pH step is performed
show significant affinity to protein A.
on a cell-free intermediate, post initial capture using protein A
chromatography.
2.1.3 log10 reduction value (LRV), n—typically used to
describe the degree of reduction of a population, in this case
1.4 The 5 log10 inactivation of murine retrovirus claimed
rodent retrovirus, by the treatment process.
by using this practice will be utilized in conjunction with other
clearance unit operations (for example, chromatography and
-1
2.1.3.1 Discussion—Each log reduction (10 ) represents a
virus retentive filtration) to assure sufficient total process
90 % reduction in the population. So a process shown to
clearance of murine retroviruses, which will be supportive of
-6
achieve a 6-log reduction (10 ) will reduce a population from
early phase regulatory filings.
a million (10 ) to 1.
1.5 The values stated in SI units are to be regarded as
2.1.4 monoclonal antibody (mAb), n—monospecific anti-
standard. No other units of measurement are included in this
bodies which have affinity for the same antigen and are made
standard.
from a master cell bank, cloned from a parent cell.
1.6 This international standard was developed in accor-
2.1.5 murine leukemia virus (MuLV), n—retroviruses named
dance with internationally recognized principles on standard-
for their ability to cause cancer in murine (mouse) hosts.
ization established in the Decision on Principles for the
2.1.5.1 Discussion—MuLV is a member of the genus Gam-
Development of International Standards, Guides and Recom-
maretrovirus. MuLV is an enveloped spherical RNA virus
mendations issued by the World Trade Organization Technical
which has a diameter of 80–110 nm and has low chemical
Barriers to Trade (TBT) Committee.
resistance. MuLV is used as a model for non-defective C-type
2. Terminology endogenous retrovirus or retrovirus like particles produced by
murine hybridoma and CHO cell lines. MuLV is used to assess
2.1 Definitions of Terms Specific to This Standard:
rodent retrovirus clearance of protein purification processes
2.1.1 IgG fusion protein, n—a dimeric protein comprised of
that use rodent cells for production.
two monomers, each monomer consisting of a peptide se-
2.1.6 recombinant protein, n—produced from the expres-
sion of recombinant DNA within living cells.
This practice is under the jurisdiction of ASTM Committee E55 on Manufac-
ture of Pharmaceutical and Biopharmaceutical Products and is the direct responsi-
2.1.6.1 Discussion—Recombinant DNA is genetically engi-
bility of Subcommittee E55.12 on Process Applications.
neered by inserting foreign DNA into the DNA of an appro-
Current edition approved Feb. 1, 2019. Published March 2019. Originally
priate host so that the foreign DNA is replicated along with the
approved in 2012. Last previous edition approved in 2012 as E2888 – 12. DOI:
10.1520/E2888-12R19. host DNA.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
E2888 − 12 (2019)
2.1.7 retrovirus, n—an RNA virus that is propagated in a depends on reactant concentration (that is, H+ ion concentra-
host cell using the reverse transcriptase enzyme to produce tion as measured by pH), time of reaction and temperature of
DNA from its RNA genome. reaction. Implementing the parameters that give robust and
2.1.7.1 Discussion—DNA is then incorporated into the effective rodent retrovirus inactivation established by this
host’s genome by an integrase enzyme. The virus is thereafter pratice, in conjunction with other clearance unit operations (for
replicated as part of the host cell’s DNA. Retroviruses are example, chromatogr
...

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3.1 Rodent-derived cell lines are widely used in the production of biopharmaceutical drugs such as mAbs and Fc fusion proteins. These cell lines have been shown to contain genes encoding endogenous retroviral-like particles or endogenous retrovirus. Despite the lack of evidence for an association between such rodent retroviruses and disease in humans, the potential contamination of human therapeutics raises safety concerns for biopharmaceutical drugs. Additionally, adventitious agents such as viruses can be introduced into a biopharmaceutical drug substance manufacturing process from other sources, and potential safety issues can be attributed to these potential unknowns. For these reasons, effective viral clearance is an essential aspect of an integrated approach combining safety testing and process characterization which ensures virus safety for biopharmaceutical drug products made using rodent cell lines.  
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1.1 This practice assures effective inactivation of ≥4 log10 of infectious rodent retrovirus (that is, reduction from 10 000 to 1 infectious rodent retrovirus or removal of 99.99 % of infectious rodent retroviruses) in the manufacturing processes of monoclonal antibodies or immunoglobulin G (IgG) Fc fusion proteins manufactured in rodent-derived cell lines that do not target retroviral antigens. Rodent retrovirus is used as a model for rodent cell substrate endogenous retrovirus-like particles potentially present in the production stream of these proteins.  
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Note 1: This test method facilitates the testing of hydrophobic surfaces by utilizing cells held in an agar slurry matrix. This test method, as written, is inappropriate to determine efficacy against biofilm cells, which are different both genetically and metabolically than planktonic cells used in this test.  
1.3 This method can confirm the presence of antimicrobial activity in plastics or hydrophobic surfaces and allows determination of quantitative differences in antimicrobial activity between untreated plastics or polymers and those with bound or incorporated low water-soluble antimicrobial agents. Comparisons between the numbers of survivors on preservative-treated and control hydrophobic surfaces may also be made.  
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1.2 This guide addresses the unique considerations that must be accounted for in the design and execution of interlaboratory studies intended to determine the precision of microbiological test methods designed to quantify microbial contamination in fuels, lubricants and similar low water-content (water activity  
1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
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5.1 Vegetative biofilm bacteria are phenotypically different from suspended planktonic cells of the same genotype. Biofilm growth reactors are engineered to produce biofilms with specific characteristics. Altering either the engineered system or operating conditions will modify those characteristics. The goal in biofilm research and efficacy testing is to choose the growth reactor that generates the most relevant biofilm for the particular study.  
5.2 The purpose of this test method is to direct a user in how to grow, treat, sample and analyze a Pseudomonas aeruginosa biofilm using the MBEC Assay. Microscopically, the biofilm is sheet-like with few architectural details as seen in Harrison et al (6). The MBEC Assay was originally designed as a rapid and reproducible assay for evaluating biofilm susceptibility to antibiotics (2). The engineering design allows for the simultaneous evaluation of multiple test conditions, making it an efficient method for screening multiple disinfectants or multiple concentrations of the same disinfectant. Additional efficiency is added by including the neutralizer controls within the assay device. The small well volume is advantageous for testing expensive disinfectants, or when only small volumes of the disinfectant are available.
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1.1 This test method specifies the operational parameters required to grow and treat a Pseudomonas aeruginosa biofilm in a high throughput screening assay known as the MBEC (trademarked)2 (Minimum Biofilm Eradication Concentration) Physiology and Genetics Assay. The assay device consists of a plastic lid with ninety-six (96) pegs and a corresponding receiver plate with ninety-six (96) individual wells that have a maximum 200 μL working volume. Biofilm is established on the pegs under batch conditions (that is, no flow of nutrients into or out of an individual well) with gentle mixing. The established biofilm is transferred to a new receiver plate for disinfectant efficacy testing.3, 4 The reactor design allows for the simultaneous testing of multiple disinfectants or one disinfectant with multiple concentrations, and replicate samples, making the assay an efficient screening tool.  
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1.7 ASTM International takes no position respecting the validity of any patent rights asserted in connection with any item mentioned in this standard. Users of this standard are expressly advised that determination of the validity of any such patent rights, and the risk of infringement of such rights, are entirely their own responsibility.  
1.8 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.  
1.9 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.

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5.1 Bacteria that exist in biofilms are phenotypically different from suspended cells of the same genotype. Research has shown that biofilm bacteria are more difficult to kill than suspended bacteria (5, 7). Laboratory biofilms are engineered in growth reactors designed to produce a specific biofilm type. Altering system parameters will correspondingly result in a change in the biofilm. For example, research has shown that biofilm grown under high shear is more difficult to kill than biofilm grown under low shear (5, 8). The purpose of this test method is to direct a user in the laboratory study of a Pseudomonas aeruginosa biofilm by clearly defining each system parameter. This test method will enable an investigator to grow, sample, and analyze a Pseudomonas aeruginosa biofilm grown under high shear. The biofilm generated in the CDC Biofilm Reactor is also suitable for efficacy testing. After the 48 h growth phase is complete, the user may add the treatment in situ or remove the coupons and treat them individually.
SCOPE
1.1 This test method specifies the operational parameters required to grow a reproducible (1)2 Pseudomonas aeruginosa ATCC 700888 biofilm under high shear. The resulting biofilm is representative of generalized situations where biofilm exists under high shear rather than being representative of one particular environment.  
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  • Standard
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ASTM
ASTM D8412-21(Guide)
Standard Guide for Quantification of Microbial Contamination in Liquid Fuels and Fuel-Associated Water by Quantitative Polymerase Chain Reaction (qPCR)

SIGNIFICANCE AND USE
5.1 This guide provides a protocol for detecting, characterizing, and quantifying nucleic acids (that is, DNA) of living and recently dead microorganisms in fuels and fuel-associated waters by means of a culture independent qPCR procedure. Microbial contamination is inferred when elevated DNA levels are detected in comparison to the expected background DNA level of a clean fuel and fuel system.  
5.2 A sequence of protocol steps is required for successful qPCR testing.  
5.2.1 Quantitative detection of microorganisms depends on the DNA-extraction protocol and selection of appropriate oligonucleotide primers.  
5.2.2 The preferred DNA extraction protocol depends on the type of microorganism present in the sample and potential impurities that could interfere with the subsequent qPCR reaction.  
5.2.3 Primers vary in their specificity. Some 16S and 18S RNA gene regions present in the DNA of prokaryotic and eukaryotic microorganisms appear to have been conserved throughout evolution and thus provide a reliable and repeatable target for gene amplification and detection. Amplicons targeting these conserved nucleotide sequences are useful for quantifying total population densities. Other target DNA regions are specific to a metabolic class (for example, sulfate reducing bacteria) or individual taxon (for example, the bacterial species Pseudomonas aeruginosa). Primers targeting these unique nucleotide sequences are useful for detecting and quantifying specific microbes or groups of microbes known to be associated with biodeterioration.  
5.3 Just as the quantification of microorganisms using microbial growth media employs standardized formulations of growth conditions enabling the meaningful comparison of data from different laboratories (Practice D6974), this guide seeks to provide standardization to detect, characterize, and quantify nucleic acids associated with living and recently dead microorganisms in fuel-associated samples using qPCR.
Note 3: Many primers, an...
SCOPE
1.1 This guide covers procedures for using quantitative polymerase chain reaction (qPCR), a genomic tool, to detect, characterize and quantify nucleic acids associated with microbial DNA present in liquid fuels and fuel-associated water samples.  
1.1.1 Water samples that may be used in testing include, but are not limited to, water associated with crude oil or liquid fuels in storage tanks, fuel tanks, or pipelines.  
1.1.2 While the intent of this guide is to focus on the analysis of fuel-associated samples, the procedures described here are also relevant to the analysis of water used in hydrotesting of pipes and equipment, water injected into geological formations to maintain pressure and/or facilitate the recovery of hydrocarbons in oil and gas recovery, water co-produced during the production of oil and gas, water in fire protection sprinkler systems, potable water, industrial process water, and wastewater.  
1.1.3 To test a fuel sample, the live and recently dead microorganisms must be separated from the fuel phase which can include any DNA fragments by using one of various methods such as filtration or any other microbial capturing methods.  
1.1.4 Some of the protocol steps are universally required and are indicated by the use of the word must. Other protocol steps are testing-objective dependent. At those process steps, options are offered and the basis for choosing among them are explained.  
1.2 The guide describes the application of quantitative polymerase chain reaction (qPCR) technology to determine total bioburden or total microbial population present in fuel-associated samples using universal primers that allow for the quantification of 16S and 18S ribosomal RNA genes that are present in all prokaryotes (that is, bacteria and archaea) and eucaryotes (that is, mold and yeast collectively termed fungi), respectively.  
1.3 This guide describes laboratory protocols. As described in Practice D7464, the...

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