Federal Register / Vol. 48, No. 44 / Friday, March 4, 1983 / Notices 
9437 
MBS. Experiments Involving More Than 
10 Liters of Culture. 'I lie appropriate 
containment will be decided by the [BC. 
Where appropriate, the large-scale 
containment recommendations of the NIH 
should be used (45 FR 2496U). 
Dr. Waitz said greater flexibility in 
scale-up procedures would be gained if 
the 1BC could be notified simultaneously 
with initiation of large-scale procedures 
involving EL coli K-12, Saccharomyces 
cerevisiae and Bacillus subtilis host- 
vector systems. He suggested that 
Section III— B— 5 be modified to read as 
follows: 
III— D— 5. Experiments Involving More Than 
10 Liters of Culture. The appropriate 
containment will be decided by the IBC 
except where exempted under Section III— D— 
5. Where appropriate, the large-scale 
containment recommendations of the NIH 
should be used (45 FR 24968). 
Language in Appendix C, Exemptions 
Under II1-D-5, would also be modified 
to reflect this change. The relevant 
paragraphs of Appendix C— II. C-riL and 
C-IV, which deal with exceptions to the 
exemption, would be modified to read 
as follows: 
Large-scale experiments (i.e_ more than 10 
liters of culture) require EBC notice 
simultaneously with the initiation of 
experiments where IBC-approved practices 
and an IBC-approved Pl-LS containment 
facility will be used. Where these conditions 
are not satisfied, refer to Part III-B-5. 
III. Proposed Modifications of Physical 
Containment Recommendations for 
Large-Scale Uses of Organisms 
Containing Recombinant DNA 
Molecules (45 FR 24968, Federal Register 
of April 11, 1980) 
A. Proposed Modification of Section 
VII-D-6 
Mr. Richard F. Geoghegan of E. L 
DuPont de Nemours and Company has 
requested that Section VII-D-8 of the 
Large-Scale Recommendations be 
revised. Section VTI-D-6 reads as 
follows: 
VII-D-6. A closed system used for the 
propagation and growth of viable organisms 
containing recombinant DNA molecules shall 
be operated so that the space above the 
culture level will be maintained at or slightly 
below atmospheric pressure. 
Mr. Geoghegan proposed that Section 
VII-D-6 be modified to read as follows: 
VII-D-6. A closed system used for the 
propagation and growth of viable organisms 
containing recombinant DNA molecules shall 
be operated so that the space above the 
culture level be maintained at no more than 
10 psig. 
He argued that the revision will 
permit fermentations more in line with 
industrial practices (high biomass 
production through efficient oxygen 
transfer) to be conducted, without 
compromising safety by operating at 
unnecessarily high vessel pressures. The 
RAC Large-Scale Review Working 
Group, at its meeting on October 26, 
1982, however, felt the language might 
appropriately be modified, but felt no 
specific pressure limit should be 
indicated as any pressure limit should 
be dependent on the maximum design 
presure of the system. The working 
group, thus, suggested the following 
language: 
VII-D-6. A closed system used for the 
propagation and growth of viable organisms 
containing recombinant DNA molecules shall 
be operated so that the space above the 
culture level will be maintained at a pressure 
as low as possible, consistent with equipment 
design, in order to maintain the integrity of 
containment features. 
The Large-Scale Review Working 
Group at its meeting on October 28, 
1982, recommended this language by a 
vote of five in favor, none opposed, and 
no abstentions. 
Both proposed modifications of 
Section VII-D-6 are published for 
comment. 
B. Proposed Modification of Sections 
VII-B-1, VII-C-1. and Vll-D-1 
Dr. Allan Waitz of DNAX Corporation 
has proposed revising Sections VII-B-1, 
VII-C-1, and VII-D-1 of the Large-Scale 
Recommendations. The current language 
of those Sections reads as follows: 
VII-B-1. Cultures of viable organisms 
containing recombinant DNA molecules shall 
be handled in a closed system (e.g., closed 
vessel used for the propagation and growth of 
cultures) or other primary containment 
equipment (e.g., biological safety cabinet 
containing a centrifuge used to process 
culture fluids) which is designed to reduce 
the potential for escape of viable organisms. 
Volumes less than 10 liters may be handled 
outside of a closed system or other primary 
containment equipment provided all physical 
containment requirements specified in 
Section II— B— 1 of the Giudelines are met. 
VII-C-1. Cultures of viable organisms 
containing recombinant DNA molecules shall 
be handled in a closed system (e.g., closed 
vessel used for the propagation and growth of 
cultures) or other primary containment 
equipment (e.g.. Class HI biological safety 
cabinet containing a centrifuge used to 
process culture fluids) which is designed to 
prevent the escape of viable organisms. 
Volumes less than 10 liters may be handled 
outside of a closed system or other primary 
containment equipment provided all physical 
containment requirements specified in 
Section II— B— 2 of the Guidelines are met. 
Vll-D-1. Cultures of viable organisms 
containing recombinant DNA molecules shall 
be handled In a closed system (e.g., closed 
vessels used for the propagation and growth 
of cultures) or other primary containment 
equipment (e.g., Class III biological safety 
cabinet containing a centrifuge used to 
process culture fluids) which is designed to 
prevent the escape of viable organisms. 
Volumes less than 10 Liters may l>e handled 
outside of a closed system provided all 
physical containment requirements specified 
in Section Il-B-3 of the Guidelines are met. 
Dr. Waitz suggested the first sentence 
in all of these sections should be 
modified to read as follows: 
Cultures of viable organisms containing 
recombinant DNA molecules shall be 
handled in a closed system used for the 
propagation, growth and processing of 
cultures, other primary containment 
equipment, or other appropriate method of 
containment approved by the IBC which is 
designed to reduce the potential for escape of 
viable organisms * * * 
The Large-Scale Review Working 
Group, at its meeting on October 26, 
1982, pointed out that a closed primary 
system is one from which there is no 
release of organisms into the 
environment or work place; a primary 
system could be defined as a fermentor 
attached to several pieces of processing 
equipment. The working group agreed 
that the language as currently written is 
purposely flexible while at the same 
time conveying the intent; they, 
therefore, recommended against 
accepting this proposal. 
C. Proposed Modification of Section 
VII-B-3 
Dt. Allan Waitz also proposed 
amending Part VTI-B-3 which reads as 
follows: 
VII-B-3. Sample collection from a closed 
system, the addition of materials to a closed 
system and the transfer of culture fluids from 
one closed system to another shall be done in 
a manner which prevents the release of 
aerosols or contamination of exposed 
surfaces. 
Dr. Waitz suggested that the word 
“prevents” implies an absolute 
condition which at the Pl-LS level is 
neither realistic nor necessai^. He 
suggested that the work "minimizes” be 
substituted for the word "prevents.” The 
Large-Scale Review Working Group did 
not discuss this proposal at its meeting 
on October 26, 1982. 
IV. Proposed Incorporation Into 
Guidelines of Physical Containment 
Recommendations for Large-Scale Uses 
of Organisms Containing Recombinant 
DNA Molecules 
Dr. Allan Waitz of DNAX Corporation 
proposed that the Physical Containment 
Recommendations for Large-Scale Uses 
of Organisms Containing Recombinant 
DNA Molecules (45 FR 24968) be 
formalized by incorporating them into 
the Guidelines as a new Appendix. Dr. 
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