Guidance for Industry 
 
Manufacturing Biological 
Intermediates and Biological Drug 
Substances Using Spore-Forming 
Microorganisms 
 
 
 
 
 
 
 
Additional copies of this guidance are available from the Office of Communication, Training and 
Manufacturers Assistance (HFM-40), 1401 Rockville Pike, Suite 200N, Rockville, MD  20852-
1448, or by calling 1-800-835-4709 or 301-827-1800, or from the Internet at 
http://www.fda.gov/cber/guidelines.htm.  
 
For questions on the content of this guidance, contact Cynthia Kelley at 301-827-0636 or John 
Eltermann at 301-827-3031.  
 
 
 
U.S. Department of Health and Human Services 
Food and Drug Administration 
Center for Biologics Evaluation and Research 
September 2007 Contains Nonbinding Recommendations 
 
Table of Contents 
 
I.  INTRODUCTION.............................................................................................................1 
II.  SCOPE...............................................................................................................................2 
III.  BACKGROUND...............................................................................................................3 
IV.  MANUFACTURING WITH SPORE-FORMERS IN A SEPARATE DEDICATED 
BUILDING........................................................................................................................4 
A.  Physical Establishment and Equipment.............................................................4 
1.  Containment........................................................................................................4 
a.  Building Construction and Configuration.......................................................4 
b.  Air Handling Units (AHUs)............................................................................4 
c.  Equipment Dedication....................................................................................5 
2.  Procedural Control..............................................................................................5 
a.  Personnel Gowning and Flows.......................................................................5 
b.  Material Transfer............................................................................................5 
c.  Equipment Cleaning........................................................................................6 
B.  Waste Disposal......................................................................................................6 
V.  MANUFACTURING WITH SPORE-FORMERS IN A MULTIPRODUCT 
MANUFACTURING BUILDING:  PROCESS CONTAINMENT.............................6 
A.  Physical Establishment and Equipment.............................................................7 
1.  Containment........................................................................................................7 
a.  Building Construction and Configuration.......................................................7 
b.  Air Handling Units (AHUs)............................................................................7 
c.  Equipment Dedication....................................................................................8 
2.  Procedural Control..............................................................................................8 
a.  Personnel Gowning and Flows.......................................................................8 
b.  Material Transfer............................................................................................8 
c.  Equipment Cleaning........................................................................................9 
B.  Waste Disposal......................................................................................................9 
C.  Campaign Changeovers........................................................................................9 
D.  Monitoring Specific for the Spore-Former.......................................................10 
1.  Environmental Monitoring Specific for the Spore-Former..............................10 
2.  Test Methods Specific to the Spore-Former.....................................................11 
a.  Specificity.....................................................................................................11 
b.  Sample Recovery..........................................................................................11 
c.  Detection Limit.............................................................................................11 
d.  Sample Locations..........................................................................................11 
VI.  SPILL CONTAINMENT...............................................................................................11 
VII.  MAINTENANCE AND DECOMMISSIONING.........................................................12 
A.  Maintenance........................................................................................................12 
i  Contains Nonbinding Recommendations 
 
B.  Decommissioning.................................................................................................12 
VIII.  DEFINITIONS................................................................................................................12 
APPENDIX:  SPORE-FORMING MANUFACTURING DIAGRAM.................................14 
ii  Contains Nonbinding Recommendations 
 
Guidance for Industry 
 
Manufacturing Biological Intermediates and Biological Drug 
Substances Using Spore-Forming Microorganisms 
 
 
This guidance represents the Food and Drug Administrations (FDAs) current thinking on this 
topic.  It does not create or confer any rights for or on any person and does not operate to bind 
FDA or the public. You can use an alternative approach if the approach satisfies the 
requirements of the applicable statutes and regulations.  If you want to discuss an alternative 
approach, contact the appropriate FDA staff.  If you cannot identify the appropriate FDA staff, 
call the appropriate number listed on the title page of this guidance. 
 
 
I.  INTRODUCTION 
 
The former regulations at 21 CFR 600.11(e)(3) ( 600.11(e)(3)) required that all work with 
spore-bearing microorganisms (spore-formers) be conducted in an entirely separate building, or 
in a completely walled-off portion of a multiproduct building.  This isolated building had to be 
dedicated exclusively for the manufacturing or storage of spore-formers.  Previously, areas in a 
multiproduct building used for manufacturing with spore-formers were required to be 
constructed to prevent cross-contamination of other areas, and were required to include entrances 
that were separate and independent from the remainder of the facility.  All equipment used for 
manufacturing spore-formers was to be permanently identified and reserved exclusively for use 
with those microorganisms.  Any materials destined for further manufacture were to be removed 
from this area only under conditions that prevented the introduction of spores into other 
manufacturing areas. 
 
We, the Food and Drug Administration (FDA), modified the regulatory requirements for the 
manufacturing of biological products with spore-formers to allow greater manufacturing 
flexibility.  Under the revised regulation1we no longer require the use of permanently dedicated 
 
buildings and equipment for spore-formers, if certain controls and precautions are applied.  We 
recognize that advances in facility, system, equipment design, testing, and sterilization 
technologies have increased the ability of manufacturers to control and analyze the manufacture 
of biological products.  As industry has gained experience with these new technologies, we 
found that manufacturers could evaluate aspects of a biological products safety and purity with 
testing.  The use of appropriate procedural controls, validated processes, and enhanced testing 
capability provide the manufacturer with a degree of confidence that their biological product 
                                                 
1 In the Federal Register of December 30, 2003, FDA published for public comment the Direct Final Rule entitled, 
Revision of the Requirements for Spore-Forming Microorganisms (68 FR 75116), and the accompanying 
Proposed Rule entitled, Revision of the Requirements for Spore-Forming Microorganisms; Companion to Direct 
Final Rule (68 FR 75179).  In the Federal Register of May 14, 2004, FDA published the Revision of the 
Requirements for Spore-Forming Microorganisms; Confirmation of Effective Date (69 FR 26768) confirming the 
effective date of June 1, 2004, for the direct final rule. 
1  Contains Nonbinding Recommendations 
 
achieves the expected levels of safety and purity.  Areas of special concern, such as process 
containment, contamination with pathogenic and/or toxic agents, sterilization, and disinfection 
can be addressed using currently available procedures and processes. 
 
We recognize that spore-formers are currently used in manufacturing processes and there will 
likely be a need to use them in the immediate future.  However, for the production of future 
biological products, manufacturers are encouraged to identify alternatives to the use of spore-
forming microorganisms whenever possible.  Such alternatives could include the use of 
sporulation deficient strains, or recombinant proteins expressed in nonspore-forming 
microorganisms.  We anticipate that certain second-generation vaccines, including those against 
anthrax, botulism, and tetanus will likely contain recombinant proteins.  The revised regulation 
uses the term spore-forming microorganisms to describe organisms that are capable of spore 
production.  The term spore-bearing microorganism, which was used in the earlier regulation, 
is not used because the term spore-forming microorganism is the more commonly accepted 
description of this class of microorganisms.  For the purposes of this guidance, the term spore-
forming microorganism (or spore-former) includes both the spore and vegetative forms of the 
organism. 
 
This guidance document finalizes the draft guidance entitled Guidance for Industry: 
Manufacturing Biological Drug Substances, Intermediates, or Products Using Spore-Forming 
Microorganisms dated February 2005 (February 24, 2005; 70 FR 9084). 
 
FDAs guidance documents, including this guidance, do not establish legally enforceable 
responsibilities.  Instead, guidance describes the agencys current thinking on a topic and should 
be viewed only as recommendations, unless specific regulatory or statutory requirements are 
cited.  The use of the word should in agency guidance means that something is suggested or 
recommended, but not required. 
 
 
II.  SCOPE 
 
The purpose of this document is to provide to you, manufacturers of biological intermediates and 
biological drug substances using spore-forming microorganisms, guidance in response to 
changes made to  600.11(e)(3).  The revised regulation describes the requirements for 
manufacturing using spore-forming microorganisms and allows manufacturers greater flexibility 
than under the prior regulation.  We are limiting the scope of this guidance to biological 
intermediates and biological drug substances as spore-formers should not be present in the 
finished dosage form (biological drug product).  Once the biological intermediates or biological 
drug substances have been separated from the viable spore-forming microorganism, the concern 
for cross-contamination diminishes.  We recommend that you establish a distinct crossover point 
in the separation process. 
 
This guidance applies to biological manufacturing processes utilizing spore-forming 
microorganisms regulated under the Federal Food, Drug and Cosmetic Act (the Act), the Public 
Health Service Act (PHS Act) and the applicable regulations in 21 CFR Parts 600 through 680.  
Section 501(a)(2)(b) of the Act requires that the methods used in, or the facilities or controls 
2  Contains Nonbinding Recommendations 
 
used for, the manufacture, processing, packing or holding of drugs, including biological 
intermediates and biological drug substances, conform to or are operated and administered in 
conformity with current good manufacturing practice to assure that such drug meets the 
requirements of the Act as to safety and has the identity and strength, and meets the quality and 
purity characteristics, which it purports or is represented to possess.  
 
This guidance does not apply to allergenic and fungi source material, in vitro diagnostics, or 
therapeutics.  Additionally, this guidance does not apply to spore-forming microorganisms used 
for the supplemental sterilization procedure control test described in  600.11(e)(2). 
 
When the spore-former used in the manufacturing process is a pathogen or is potentially 
pathogenic, the stricter requirements related to equipment and supplies between  600.11(e)(5) 
and revised  600.11(e)(3) must be applied. 
 
 
III.  BACKGROUND 
 
Spore-formers are used in the production of certain biological products.  Biological intermediates 
or biological drug substances derived from spore-formers may be used as source material for 
further manufacture into final biological drug products, such as vaccines.  Bacteria produce 
spores as a means to survive adverse environmental conditions.  In general, spores show 
enhanced resistance over bacteria to high temperatures, freezing, dryness, antibacterial agents, 
radiation, and toxic chemicals.  Under favorable conditions, spores can germinate into actively 
growing vegetative bacterial cells.  Some of these spore-formers are human pathogens and are 
associated with high morbidity and mortality.  Due to their unique survival properties, spore-
formers pose great challenges to manufacturers. In order to ensure the safety of a product 
manufactured in a facility in which spore-formers are present, these microorganisms must be 
kept under stringent control in order to avoid the release of spores into the manufacturing area 
where they have the potential to cross-contaminate other products and areas. ( 600.11(e)(3)(i)). 
 
Manufacturing with spore-formers requires varying levels of control depending on the 
characteristics (e.g., virulence toward humans) of the microorganism that is utilized in a 
manufacturing process.  The level of confidence a specific process provides for safety and purity 
may also be a factor in deciding the level of manufacturing control necessary.  You must institute 
the appropriate controls in the manufacturing facility, associated equipment, and manufacturing 
procedures to avoid contamination.  ( 600.11(a), 600.11(e)(3)(i), 600.11(e)(5)).   The 
recommendations described in this guidance are intended for manufacturing with the type of 
spore-formers that are currently used by the regulated industry and included in the scope of this 
guidance.  Any atypical or novel spore-formers may require specific controls that are unique for 
that particular microorganism. 
 
Prevention of spore contamination can be achieved by using a separate dedicated building or by 
using process containment if manufacturing is conducted in a multiproduct manufacturing 
building.  Containment is established by facility design and/or the use of procedures and 
equipment that prevent the release of spores into adjacent areas.  ( 600.11(e)(3)). 
 
3  Contains Nonbinding Recommendations 
 
 
IV.  MANUFACTURING WITH SPORE-FORMERS IN A SEPARATE DEDICATED 
BUILDING 
 
This section applies if you choose to use a separate dedicated building configuration for 
manufacturing with spore-formers.  Although you are no longer required to use this type of 
configuration, if you do so, you must follow the applicable requirements in  600.11(e)(3)(i) to 
avoid having to satisfy the process containment requirements in subsection (e)(3)(ii).  This 
section of the guidance document reiterates the requirements in subsection (e)(3)(i) applicable to 
separate dedicated buildings, and provides additional recommendations for using this 
configuration. 
 
A.  Physical Establishment and Equipment 
 
The use of a separate dedicated building for manufacturing processes involving spore-
formers is the simplest means to prevent cross-contamination.  The separate dedicated 
building configuration is not intended to accommodate the manufacturing of multiple 
products using spore-formers.  If multiple products are manufactured in the same area or 
within the same building using spore-formers, then additional criteria will apply.  See  
 600.11(e)(3)(ii) and section V. below. 
 
1.  Containment 
 
a.  Building Construction and Configuration 
 
If you choose to use a separate dedicated building configuration for 
manufacturing processes involving spore-formers, your building must be 
constructed to prevent contamination. ( 600.11(e)(3)(i)). Your separate dedicated 
building should be constructed with walls that extend to the ceiling with properly 
sealed joints at wall/ceiling intersections, separate heating, ventilation, and air 
conditioning (HVAC), water drops, sewer line, and an independent entrance 
thereby containing the building from surrounding areas.   
 
We recommend that all surfaces be solid, hard, non-porous, and cleanable, 
including ceilings and walls.  To the extent you are using aseptic processing, 
floors, walls, and ceilings should have smooth, hard surfaces that are easily 
cleanable.2  
 
b.  Air Handling Units (AHUs) 
 
AHUs should not be shared with other buildings.  Dedicated re-circulating AHUs 
are acceptable within the separate dedicated building.  We recommend that 
exhaust not be located near other AHU intakes.  We also recommend that you 
                                                 
2 Please refer to FDAs guidance for industry entitled Sterile Drug Products Produced by Aseptic Processing  
Current Good Manufacturing Practice, dated September 2004 (October 4, 2004, 69 FR 59258) 
(http://www.fda.gov/cder/guidance/5882fnl.htm), for the agencys current thinking on aseptic processing. 
4  Contains Nonbinding Recommendations 
 
consider using high efficiency particulate air (HEPA) filtration of exhaust, and 
that you maintain the building at negative pressure with respect to the outside 
environment and/or adjacent areas. 
 
c.  Equipment Dedication 
 
If you choose a separate dedicated building configuration, major equipment 
should be identified as used for manufacturing with the particular spore-former.  
We recommend that you dedicate all equipment. 
 
2.  Procedural Control 
 
a.  Personnel Gowning and Flows 
 
Personnel who work in manufacturing using spore-formers must complete an 
outer covering change or wear protective covering prior to entering areas where 
other products are manufactured ( 600.10(c)(3)).  We also recommend that 
personnel shower and complete a clean clothing change prior to entering other 
areas or interacting with personnel not directly involved in the manufacturing of 
spore-formers. 
 
b.  Material Transfer 
 
Any material transferred out of a spore-former manufacturing area that may cross-
contaminate other products, product containers, intermediates, or materials used 
in the manufacturing of other products should be decontaminated via a 
decontamination chamber prior to exiting.  If a decontamination chamber is not 
feasible, then a series of airlocks utilizing surface decontamination with an 
appropriate agent may be acceptable. 
 
When utilizing a decontamination chamber system such as an autoclave, sterilize-
in-place (SIP) cycle, or gas chamber, the decontamination cycle should be 
validated using methodologies that ensure that the spore-former would be 
inactivated.  We recommend that you incorporate a worst-case approach.  You 
should not use the same chamber for both the decontamination of the spore-
former and the sterilization of other production items. 
 
When it is necessary to employ an airlock system for decontamination of material 
that is transferred out of a manufacturing area, your procedures should provide for 
the decontamination process.  Use of a liquid or gaseous agent to inactivate the 
spore-formers and decontamination agent efficacy studies are recommended.  We 
recommend that you incorporate a worst-case approach. We also recommend that 
you consider parameters that include the composition of the material, exposure 
time, concentration, pH, temperature and any other relevant factors.  If the airlock 
system involves only local decontamination of the equipment or item, such as 
wipe downs, then we recommend that the decontamination process be conducted 
5  Contains Nonbinding Recommendations 
 
and validated in series (multiple airlocks) to ensure that no trailing contamination 
is encountered.  We recommend that inside personnel pass the outgoing material 
and/or equipment into an interior material airlock (i.e., Material Airlock (Primary 
Decontamination) as shown in the Appendix), where they decontaminate the 
items according to approved validated procedures.  After initial decontamination, 
the inside personnel transfer the outgoing items into an exterior material airlock 
(i.e., Material Airlock (Secondary Decontamination) as shown in the Appendix).  
The outside personnel then provide a second decontamination step (also 
validated) on the outgoing items, and then transfer the material and/or equipment 
into an exit corridor or room.  We recommend that temporal separation between 
personnel be maintained and that personnel not use material airlocks for exiting 
(see Appendix). 
 
c.  Equipment Cleaning 
 
No special equipment cleaning is needed, beyond what  600.11(b) requires, 
when manufacturing with spore-formers, provided that the equipment is 
maintained within the dedicated processing area. 
 
B.  Waste Disposal 
 
Process waste can be treated in a similar fashion as the material transferred out of the 
manufacturing area or processed through a validated bio-waste system. When waste is 
transferred out of the manufacturing area, we recommend that it be transferred out in 
biohazard bags via multiple airlocks in a manner similar to the material transfer process 
described in section IV.A.2.b.  We recommend that waste be bagged in the processing 
room and transferred to the interior material airlock where the outside of the bag is 
decontaminated by the inside personnel. After the initial decontamination, inside 
personnel places the bagged waste into another biohazard bag and transfers it into the 
exterior material airlock.  Outside personnel decontaminates the outside of the second 
biohazard bag and removes the double-bagged waste from the exterior material airlock 
(see Appendix). 
 
All waste must be disposed of in a manner that is in compliance with federal, state, and 
local environmental laws. 
 
 
V.  MANUFACTURING WITH SPORE-FORMERS IN A MULTIPRODUCT 
MANUFACTURING BUILDING:  PROCESS CONTAINMENT 
 
Process containment is designed to isolate equipment or an area that involves manufacturing 
using spore-forming microorganisms mechanically (Containment) and procedurally (Procedural 
Control).  This manufacturing area, including biological intermediates or biological drug 
substances, equipment, or material storage in that area, must not be used for any other purpose 
during the processing period ( 600.11(e)(3)(ii)).  The manufacturing area should contain 
adequate space for the orderly placement of equipment.  If you intend to use an area for 
6  Contains Nonbinding Recommendations 
 
manufacturing spore-formers and other products on a campaign basis, then you must develop and 
implement procedures to demonstrate containment and decontamination before initiating 
processing of other products in that manufacturing area ( 600.11(e)(3)(ii)).  These procedures 
must, at a minimum, provide for containment, environmental monitoring specific for the spore-
former, cleaning, decontamination, and movement of materials in and out of that area  
( 600.11(e)(3)(ii)).  We recommend that you implement campaign changeover procedures 
whenever an area that was previously used for a spore-forming microorganism will subsequently 
be used for manufacturing other products. 
 
A.  Physical Establishment and Equipment 
 
1.  Containment 
 
a.  Building Construction and Configuration 
 
We recommend that the manufacturing areas used for spore-forming 
microorganisms have double serial entry, exit, and material airlocks to minimize 
the potential for cross-contamination via personnel, materials, and air turbulence.  
As shown in the Appendix, the interior airlocks would serve as an air sink for the 
primary decontamination step.  The three air sinks shown in the Appendix include 
the final gowning step (Gown-in (Final)), the gross de-gowning step (Gown-out 
(Initial)), and the interior material airlock (Material Airlock (Primary 
Decontamination)).  The exterior airlocks would serve as an air dome for the 
preliminary gowning step.  The air domes shown in the Appendix include primary 
decontamination (Gown-in (Preliminary)), the final de-gowning step (Gown-out 
(Final)), and the secondary decontamination step (Material Airlock (Secondary 
Decontamination)). 
 
We recommend that all surfaces be solid, hard, non-porous, and cleanable, 
including ceilings and walls.  To the extent you are using aseptic processing, 
floors, walls, and ceilings should have smooth, hard surfaces that are easily 
cleanable.  We recommend that the final ceiling and wall finishes minimize air 
leaks. 
 
Your procedures should provide for the decontamination of all tables, shelving, 
and storage apparatus. 
 
b.  Air Handling Units (AHUs) 
 
We recommend that AHUs using 100% single-pass air be used within the area 
where manufacturing with spore-forming microorganisms occurs and that the area 
be maintained at a negative air pressure to all surrounding areas, including 
ceilings and mechanical spaces.  We recommend that exhaust not be located near 
other AHU intakes, and that HEPA filtration of exhaust be considered. 
7  Contains Nonbinding Recommendations 
 
 
c.  Equipment Dedication 
 
Wherever possible, we recommend that major processing equipment be dedicated 
for a specific product use and that such equipment be identified to show the 
specific equipment used in the manufacture of each batch of product.  If 
equipment is dedicated for a specific manufacturing process using a spore-former, 
it must be decontaminated prior to the introduction of a new production process  
( 600.11(e)(3)(ii)), and we recommend removing it from the area, where 
feasible. We recommend that dedicated equipment be segregated from other 
equipment while in storage. 
 
We recommend that small ancillary equipment and administrative items such as 
pens, logbooks, pipettes, miscellaneous glassware, and standard operating 
procedures (SOPs) be dedicated or made disposable.  Dedicated and disposable 
items must be decontaminated and/or removed from the processing area during 
the area changeover procedure ( 600.11(e)(3)(ii)). 
 
When equipment dedication and removal is not feasible, such as with large fixed 
mounted bioreactors, decontamination and cleaning procedures must be in place 
( 600.11(e)(3)(ii)).  See also section V.A.2.c. below. 
 
2.  Procedural Control 
 
a.  Personnel Gowning and Flows 
 
We recommend that personnel entry and exit flows be unidirectional.  We also 
recommend that personnel gown with multiple layers so that personnel gowning 
and de-gowning occurs in multiple stages (see Appendix) and that gowning 
procedures are designed to prevent or reduce the potential for personnel to carry 
the spore-former out of the area via their hair, skin, shoes, jewelry, or clothing.  
We recommend that disposable gowning be decontaminated using a validated 
process or adequately bagged and sealed prior to leaving the area used for spore-
forming microorganisms.  Personnel who work in manufacturing using spore-
formers must complete an outer covering change or wear protective covering 
prior to entering areas where other products are manufactured ( 600.10(c)(3)).  
We also recommend that personnel shower and complete a clean clothing 
change prior to entering other areas or interacting with personnel not directly 
involved in the manufacturing of spore-formers.  
 
b.  Material Transfer 
 
The recommendations for material transfer in a multiproduct manufacturing 
building are the same as those for a separate dedicated building (see section 
IV.A.2.b.). 
8  Contains Nonbinding Recommendations 
 
 
c.  Equipment Cleaning 
 
When the use of dedicated or disposable equipment is not feasible, such as with 
large or fixed bioreactors, decontamination and cleaning procedures must be in 
place ( 600.11(e)(3)(ii)).  In development of these procedures for each piece of 
equipment you should consider the following: 
  efficacy of the decontamination; 
  analysis of potential residue for the specific spore-former;  
  evaluation of product contact surface areas, and the interior and 
exterior areas on equipment to include controls, valves, seals, probes, 
motors, wiring harnesses, and miscellaneous external surfaces; 
  sampling procedures; and 
  analytical methods, including sensitivity of those methods. 
 
We recommend the dismantling and inspection of such equipment between 
different products.  Some cleaning agents (e.g., NaOH) can lead to pitting of 
equipment and may protect spore-formers from the decontaminant. 
 
B.  Waste Disposal 
 
The recommendations for waste disposal in a multiproduct manufacturing building are 
the same as those for a separate dedicated building (see section IV.B.). 
 
C.  Campaign Changeovers 
 
Campaign changeovers involve the cleaning and decontamination of a specific area that 
has been exposed to a spore-former in preparation for the introduction of another product 
or process into that same area. The decontamination and cleaning would include all 
equipment and items that may have been exposed to the spore-forming microorganism.  
This includes Biological Safety Cabinets and re-circulating AHUs.  In addition, 
consideration may be given to decontamination of the entire manufacturing area with a 
gaseous sterilant such as chlorine dioxide or hydrogen peroxide. 
 
Prior to introducing a new product into a facility previously used to manufacture spore-
formers, you must develop and implement procedures that focus on issues related to 
containment, environmental monitoring specific for the spore-former, cleaning, 
decontamination, and movement of equipment and materials in and out of that area  
( 600.11(e)(3)(ii)).  Campaign changeovers as outlined in this document occur when an 
area that was previously used for spore-former manufacturing will subsequently be used 
for manufacturing any other product. 
9  Contains Nonbinding Recommendations 
 
We recommend that the following steps be taken in the order indicated below for 
cleaning and decontamination of an area that is to be used on a campaign changeover: 
 
1)  All waste in the area is removed or sent to the bio-waste system, as required by 
 600.11(e)(3)(ii); 
 
2)  Stay-in-place equipment is decontaminated, as required by  600.11(e)(3)(ii); 
 
3)  Removable dedicated equipment and ancillary items are decontaminated and 
removed or sent to waste for disposal from the area according to material 
transfer procedures, as required by  600.11(e)(3)(ii) and as outlined with 
additional specific recommendations in section IV.A.2.b.; 
 
4)  Room and room fixtures (e.g., shelves, incubators, storage units) are 
decontaminated and cleaned according to procedures, as required by  
   600.11(e)(3)(ii); 
 
5)  Stay-in-place processing equipment is dismantled, cleaned, and sterilized (if 
applicable); 
 
6)  Environmental monitoring specific for the spore-formers used in 
manufacturing is performed, as required by  600.11(e)(3)(ii) (see section V.D. 
for additional recommendations); and 
 
7)  A quality control unit review of the campaign changeover data (including 
environmental monitoring results) and a quality control unit area inspection are 
executed prior to releasing the area for the next product. 
 
D.  Monitoring Specific for the Spore-Former 
 
1.  Environmental Monitoring Specific for the Spore-Former 
 
Environmental monitoring3 that is specific for the spore-former must be conducted in 
adjacent areas during manufacturing operations and in the manufacturing area after 
completion of cleaning and decontamination, at the conclusion of the campaign 
changeover procedures ( 600.11(e)(3)(ii)).  During operations, we suggest that the 
sampling and testing be conducted in the adjacent areas at the beginning, middle, and 
end of each manufacturing shift.  All areas must be free from contamination with the 
spore-former. ( 600.11(e)(3)(i)). 
                                                 
3 Please refer to FDAs Guidance for Industry:  Sterile Drug Products Produced by Aseptic Processing  Current 
Good Manufacturing Practice dated September 2004 (October 4, 2004, 69 FR 59258) 
(http://www.fda.gov/cber/gdlns/steraseptic.htm), for the agencys current thinking on aseptic processing and 
environmental monitoring. 
10  Contains Nonbinding Recommendations 
 
 
2.  Test Methods Specific to the Spore-Former 
 
Test methods used for monitoring specific for the spore-former should be qualified 
for specificity, sample recovery, and detection limits, at a minimum.  A quantitative 
or qualitative test is acceptable provided the test can be appropriately qualified. 
 
a.  Specificity 
 
Testing should be able to detect the specific spore-former and identify it in the 
presence of other microorganisms.  
 
b.  Sample Recovery 
 
Sampling qualifications should demonstrate the percent recovery of the specific 
spore-former.  
 
c.  Detection Limit 
 
Limits of detection using the specific spore-former should be established.  
 
d.  Sample Locations 
 
We recommend the adjacent areas, associated with the isolated processing area, 
be sampled near or at the points of possible egression, such as doorways, 
windows, and/or other openings. 
 
We recommend that the number of samples for testing at the conclusion of the 
campaign changeover be developed by a matrix approach and take into account 
the size and complexity of the equipment, room, and room fixtures.  We suggest 
that the exact sampling sites be determined based on equipment and items that are 
the most difficult to clean and decontaminate and the likelihood of product 
impact.  For example, controls over a formulation tank should be tested, but the 
floor in an isolated corner may not need to be included. 
 
 
VI.  SPILL CONTAINMENT 
 
We recommend that procedures be in place to address emergency responses for all spills 
involving spore-forming microorganisms regardless of location.  These procedures should 
include validated processes for containing the spill, and cleaning and decontaminating the area 
and equipment affected by the spill.  
 
11  Contains Nonbinding Recommendations 
 
VII.  MAINTENANCE AND DECOMMISSIONING 
 
A.  Maintenance 
 
We recommend that maintenance activities be performed under appropriate controls to 
prevent contamination of other products and areas within the site.  If it is necessary to 
break the integrity of the process containment to perform maintenance activities, then you 
must follow the established decontamination procedures prior to the maintenance 
activities and after all manufacturing activity has ceased. ( 600.11(e)(3)(ii)). 
 
B.  Decommissioning 
 
Decommissioning of a dedicated spore-former manufacturing area or building must 
involve a comprehensive decontamination plan with extensive spore-former specific 
testing to ensure the adequate removal and/or cleaning of the area and building prior to 
the introduction of another product (see section V.D.). ( 600.11(e)(3)(ii)). 
 
 
VIII.  DEFINITIONS 
 
a.  Air handling unit (AHU)  A component of the HVAC system that includes the fans, filters, 
coils, and other materials used to generate conditioned air. 
 
b.  Biological drug product  A finished dosage form that contains an active biological drug 
ingredient generally, but not necessarily, in association with inactive ingredients. 
 
c.  Biological drug substance  Any substance or mixture of substances intended to be used in the 
manufacture of a biological drug product and that, when used in the production of a biological 
drug product, becomes an active ingredient of the biological drug product.  Such biological 
substances are applicable to the prevention, treatment, or cure of a disease or condition of 
human beings. 
 
d.  Biological intermediates  A material produced during steps of the processing that must 
undergo further molecular change or purification before it becomes a biological drug 
substance.  Biological intermediates may or may not be isolated. 
 
e.  Biological product  Any virus, therapeutic serum, toxin, antitoxin, vaccine, blood, blood 
component or derivative, allergenic product, or analogous product, or arsphenamine or 
derivative of arsphenamine, applicable to the prevention, treatment, or cure of disease or 
condition of human beings. 
 
f.  Campaign changeovers  The process of cleaning and decontaminating a specific area, 
equipment, system, waste, and/or ancillary items exposed to a spore-forming microorganism 
in preparation for the introduction of another product or process into that same area. 
 
12  Contains Nonbinding Recommendations 
 
g.  Containment  Physical/mechanical barriers and/or systems designed to control spore-formers 
to prevent contamination. 
 
h.  Cross-contamination  Contamination of a material or product with another material or 
product. 
 
i.  Crossover point  The processing point where viable organisms are not part of the process or 
process solutions. 
 
j.  Decontamination of spore-forming microorganisms  A process that eliminates or inactivates 
viable bioburden and any associated toxic material via the use of chemical and/or physical 
means. 
 
k.  Facility  Physical structure. 
 
l.  Manufacturing area  A specified location within a facility associated with the manufacturing 
of any one product or multiple products. 
 
m.  Manufacturing facility  The physical structure associated with the manufacturing of any one 
product or multiple products. 
 
n.  Manufacturing site  The entire complex of buildings, connected or separate, and belonging 
to one entity engaged in the manufacturing of any one product or multiple products. 
 
o.  Multiproduct  More than one approved product, licensed product, investigational new drug 
product or separate process. 
 
p.  Procedural control  Manufacturing procedures executed in such a manner as to prevent or 
minimize spore-former contamination. 
 
q.  Process containment  A system designed to mechanically and procedurally isolate equipment 
or an area that involves manufacturing using spore-forming microorganisms. 
 
r.  Qualification  Action of proving and documenting that equipment, ancillary systems, or areas 
are properly designed, installed, work correctly, and actually lead to the expected results.  
Qualification is part of validation, but the individual qualification steps alone do not constitute 
process validation. 
 
s.  Spore-forming microorganism  Organisms that are capable of spore production, including 
both spore and vegetative forms of the organism. 
 
t.  Validation  Establishing documented evidence that provides a high degree of assurance that a 
specific process will consistently produce a result meeting its predetermined specifications 
and quality attributes. 
13  Contains Nonbinding Recommendations 
APPENDIX:  SPORE-FORMING MANUFACTURING DIAGRAM 
= Airflow direction 
Gown-out 
(Final) 
Pass-through 
decontamination 
chamber 
Gown-out 
(Initial) 
(Preliminary) Gown-in  (Final)Gown-in  Processing/Laboratory  Material Airlock PrimaryDecontamination)  Material Airlock SecondaryDecontamination)
( (
Mechanical Space 
14