ASTM E3042-16
(Practice)Standard Practice for Process Step to Inactivate Rodent Retrovirus with Triton X-100 Treatment
Standard Practice for Process Step to Inactivate Rodent Retrovirus with Triton X-100 Treatment<rangeref></rangeref >
SIGNIFICANCE AND USE
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.
3.2 Solvent/detergent inactivation has been widely used for decades to inactivate enveloped viruses in blood plasma derived biopharmaceutical therapies (1-3).3 Solvent/detergent systems using the detergents Triton X-100 or Polysorbate 80 along with the organic solvent tri(n-butyl)phosphate (TNBP) have been used to inactivate enveloped viruses by disrupting the viral envelope thereby reducing the ability of the enveloped virus to attach to and then infect the host cell (4 and 5).
3.3 Most manufacturers of mAbs, recombinant proteins, and Fc fusion proteins have focused on viral inactivation methods using the detergent Triton X-100 or Polysorbate 80 in the absence of TNBP (6), which can interfere with subsequent bioprocessing steps. The ability of the detergents alone to inactivate retroviruses has been demonstrated in monoclonal antibodies produced in rodent-derived cell lines (6-9). At a 2011 workshop devoted to viral clearance steps used in bioprocessing (7), investigators from one firm showed incubation with 0.2 % Triton X-100 for 60 min of hold time at a...
SCOPE
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.
1.2 The parameters specified for this practice are clarification, Triton X-100 detergent concentration, hold time, pH, and inactivation temperature.
1.3 This practice can be used in conjunction with other clearance or inactivation unit operations that are orthogonal to this inactivation mechanism to achieve sufficient total process clearance or inactivation of rodent retrovirus.
1.4 This detergent inactivation step is performed on a clarified, cell-free intermediate of the monoclonal antibody or IgG Fc fusion protein.
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 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.
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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: E3042 − 16
Standard Practice for
Process Step to Inactivate Rodent Retrovirus with Triton
1,2
X-100 Treatment
This standard is issued under the fixed designation E3042; 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 2.1.1 clarified, cell free intermediate, n—in-process pool
located downstream of the cell clarification unit operation(s),
1.1 This practice assures effective inactivation of ≥4 log of
10
which should include a filtration step of ≤0.2 μm nominal pore
infectious rodent retrovirus (that is, reduction from 10 000 to 1
size, and upstream of the initial purification step in the
infectious rodent retrovirus or removal of 99.99 % of infectious
purification process of a monoclonal antibody or IgG Fc fusion
rodent retroviruses) in the manufacturing processes of mono-
protein.
clonal antibodies or immunoglobulin G (IgG) Fc fusion pro-
2.1.1.1 Discussion—Cell clarification unit operations are
teins manufactured in rodent-derived cell lines that do not
performed on the cell culture supernatant. Cell clarification
target retroviral antigens. Rodent retrovirus is used as a model
unit operations can be one or more of the following opera-
for rodent cell substrate endogenous retrovirus-like particles
tion(s): microfiltration, centrifugation, depth filtration, or
potentially present in the production stream of these proteins.
flocculation, or combination thereof. The primary purpose of
1.2 The parameters specified for this practice are
cell clarification unit operation(s) is to remove cells used to
clarification, Triton X-100 detergent concentration, hold time,
generate monoclonal antibody or IgG Fc fusion protein and
pH, and inactivation temperature.
some proportion of cellular debris from the cell culture
1.3 This practice can be used in conjunction with other supernatant before the initial purification step. All clarification
steps must include ≤0.2 μm nominal pore size filtration to
clearance or inactivation unit operations that are orthogonal to
this inactivation mechanism to achieve sufficient total process minimize the presence of virus aggregates, prior to detergent
inactivation. Freezing or prolonged storage between ≤0.2 μm
clearance or inactivation of rodent retrovirus.
filtration and detergent inactivation should be avoided.
1.4 This detergent inactivation step is performed on a
2.1.2 enveloped virus, n—viruses in which the nucleic acid
clarified, cell-free intermediate of the monoclonal antibody or
component of the virus is surrounded by a lipid containing
IgG Fc fusion protein.
envelope acquired from the host cell during virus assembly and
1.5 The values stated in SI units are to be regarded as
budding.
standard. No other units of measurement are included in this
2.1.2.1 Discussion—Some examples of enveloped viruses
standard.
are from the families orthomyxoviridae (influenza), paramyxo-
1.6 This standard does not purport to address all of the
viridae[mumps and measles], retroviridae[human immunode-
safety concerns, if any, associated with its use. It is the
ficiency virus (HIV) and murine leukemia virus (MuLV)], and
responsibility of the user of this standard to establish appro-
herpesviridae [human herpes virus (HHV), varicella-zoster
priate safety and health practices and determine the applica-
virus (VZV), and pseudorabies virus (PRV)].
bility of regulatory limitations prior to use.
2.1.3 hold time, n—amount of time, after sufficient mixing
2. Terminology takes place, that the biological drug intermediate and retrovirus
interact with a specific chemical, in this case, the amount of
2.1 Definitions of Terms Specific to This Standard:
time the biological drug intermediate and retrovirus interact
with the Triton X-100.
1
This practice is under the jurisdiction of ASTM Committee E55 on Manufac-
2.1.3.1 Discussion—Demonstration of sufficient mixing is
ture of Pharmaceutical and Biopharmaceutical Products and is the direct responsi-
bility of Subcommittee E55.12 on Process Applications. the responsibility of the manufacturer.
Current edition approved Sept. 1, 2016. Published September 2016. DOI:
2.1.4 immunoglobulin G, IgG, n—antibody molecule com-
10.1520/E3042-16.
2
posed of four peptide chains—two gamma heavy chains and
Triton X-100 is a trademark of The Dow Chemical Company, Midlands,
Michigan, http://www.dow.com. The sole source of manufacture of the material
two light chains.
known to the committee at this time is The Dow Chemical Co
...
NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
Designation: E3042 − 16
Standard Practice for
Process Step to Inactivate Rodent Retrovirus with Triton
1,2
X-100 Treatment
This standard is issued under the fixed designation E3042; 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 2.1.1 clarified, cell free intermediate, n—in-process pool
located downstream of the cell clarification unit operation(s),
1.1 This practice assures effective inactivation of ≥4 log of
10
which should include a filtration step of ≤0.2 µm nominal pore
infectious rodent retrovirus (that is, reduction from 10 000 to 1
size, and upstream of the initial purification step in the
infectious rodent retrovirus or removal of 99.99 % of infectious
purification process of a monoclonal antibody or IgG Fc fusion
rodent retroviruses) in the manufacturing processes of mono-
protein.
clonal antibodies or immunoglobulin G (IgG) Fc fusion pro-
2.1.1.1 Discussion—Cell clarification unit operations are
teins manufactured in rodent-derived cell lines that do not
performed on the cell culture supernatant. Cell clarification
target retroviral antigens. Rodent retrovirus is used as a model
unit operations can be one or more of the following opera-
for rodent cell substrate endogenous retrovirus-like particles
tion(s): microfiltration, centrifugation, depth filtration, or
potentially present in the production stream of these proteins.
flocculation, or combination thereof. The primary purpose of
1.2 The parameters specified for this practice are
cell clarification unit operation(s) is to remove cells used to
clarification, Triton X-100 detergent concentration, hold time,
generate monoclonal antibody or IgG Fc fusion protein and
pH, and inactivation temperature.
some proportion of cellular debris from the cell culture
supernatant before the initial purification step. All clarification
1.3 This practice can be used in conjunction with other
clearance or inactivation unit operations that are orthogonal to steps must include ≤0.2 µm nominal pore size filtration to
minimize the presence of virus aggregates, prior to detergent
this inactivation mechanism to achieve sufficient total process
clearance or inactivation of rodent retrovirus. inactivation. Freezing or prolonged storage between ≤0.2 µm
filtration and detergent inactivation should be avoided.
1.4 This detergent inactivation step is performed on a
2.1.2 enveloped virus, n—viruses in which the nucleic acid
clarified, cell-free intermediate of the monoclonal antibody or
component of the virus is surrounded by a lipid containing
IgG Fc fusion protein.
envelope acquired from the host cell during virus assembly and
1.5 The values stated in SI units are to be regarded as
budding.
standard. No other units of measurement are included in this
2.1.2.1 Discussion—Some examples of enveloped viruses
standard.
are from the families orthomyxoviridae (influenza), paramyxo-
1.6 This standard does not purport to address all of the
viridae[mumps and measles], retroviridae[human immunode-
safety concerns, if any, associated with its use. It is the
ficiency virus (HIV) and murine leukemia virus (MuLV)], and
responsibility of the user of this standard to establish appro-
herpesviridae [human herpes virus (HHV), varicella-zoster
priate safety and health practices and determine the applica-
virus (VZV), and pseudorabies virus (PRV)].
bility of regulatory limitations prior to use.
2.1.3 hold time, n—amount of time, after sufficient mixing
2. Terminology takes place, that the biological drug intermediate and retrovirus
interact with a specific chemical, in this case, the amount of
2.1 Definitions of Terms Specific to This Standard:
time the biological drug intermediate and retrovirus interact
with the Triton X-100.
1
This practice is under the jurisdiction of ASTM Committee E55 on Manufac-
2.1.3.1 Discussion—Demonstration of sufficient mixing is
ture of Pharmaceutical and Biopharmaceutical Products and is the direct responsi-
the responsibility of the manufacturer.
bility of Subcommittee E55.12 on Process Applications.
Current edition approved Sept. 1, 2016. Published September 2016. DOI:
2.1.4 immunoglobulin G, IgG, n—antibody molecule com-
10.1520/E3042-16.
2
posed of four peptide chains—two gamma heavy chains and
Triton X-100 is a trademark of The Dow Chemical Company, Midlands,
Michigan, http://www.dow.com. The sole source of manufacture of the material
two light chains.
known to the committee at this time is The Dow Chemical Company. If you are
2.1.4.1 Discussion—Each IgG has two antigen binding
aware of alternative suppliers, please provide this information
...
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