I IIIII IIIIIIII II llllll lllll lllll lllll lllll lllll lllll lllll lllll lllll lllll 111111111111111111
`US 20030003014Al
`
`(19) United States
`c12) Patent Application Publication
`Metzner et al.
`
`(10) Pub. No.: US 2003/0003014 Al
`Jan. 2, 2003
`( 43) Pub. Date:
`
`(54) USE OF A HYDROGEN PEROXIDE PIASMA
`STERILIZATION METHOD FOR THE MILD
`STERILIZATION OF
`TEMPERATURE-SENSITIVE PRODUCTS
`
`(76)
`
`Inventors: Hubert Metzner, Marburg (DE); Joerg
`Lemmer, Ebsdorfergrund (DE); Horst
`Naumann, Marburg (DE)
`
`(30)
`
`Foreign Application Priority Data
`
`Jan. 26, 2001
`
`(DE) ..................................... 101 03 706.6
`
`Publication Classification
`
`Int. CI.7 ....................................................... A61L 2/20
`(51)
`(52) U.S. CI .
`................................................. 422/29; 422/33
`
`Correspondence Address:
`Finnegan, Henderson, Farabow,
`Garrett & Dunner, L.L.P.
`1300 I Street, N.W.
`Washington, DC 20005-3315 (US)
`
`(21)
`
`Appl. No.:
`
`10/052,469
`
`(22)
`
`Filed:
`
`Jan.23,2002
`
`(57)
`
`ABSTRACT
`
`The invention relates to a method for hydrogen peroxide
`plasma sterilization, wherein the chamber temperature is set
`at less than 39° C. throughout, and containers with tempera(cid:173)
`ture-sensitive products can be efficiently sterilized without
`the temperature-sensitive products showing a significant
`loss of activity or degradation.
`
`Regeneron Exhibit 1018.001
`
`

`

`Patent Application Publication
`
`Jan.2,2003
`
`US 2003/0003014 Al
`
`I
`I l
`v··
`I
`I
`I I
`I
`
`!
`
`I
`
`-~,\
`
`,I
`
`. co
`
`fl . . .. .. . .
`
`rLi-·- ·
`. - .1 -~ - -1 - - - ~~-~~~--~-..,'"'<.>=" i
`
`1
`
`Regeneron Exhibit 1018.002
`
`

`

`US 2003/0003014 Al
`
`Jan. 2, 2003
`
`1
`
`USE OF A HYDROGEN PEROXIDE PLASMA
`STERILIZATION METHOD FOR THE MILD
`STERILIZATION OF TEMPERATURE-SENSITIVE
`PRODUCTS
`
`[0001] The invention relates to a method for hydrogen
`peroxide plasma sterilization, wherein the chamber tempera(cid:173)
`ture is set at less than 39° C. throughout, and containers with
`temperature-sensitive products can be efficiently sterilized
`without the temperature-sensitive products showing a sig(cid:173)
`nificant loss of activity or degradation.
`
`BACKGROUND or THE INVENTION
`[0002] The external sterilization of pharmaceutical con(cid:173)
`tainers which contain products which are sensitive to tem(cid:173)
`perature effects or irradiation is a general problem which has
`not yet been satisfactorily solved.
`
`[0003] Autoclaving is virtually always unsuitable for bio(cid:173)
`logical products because even the most stable products
`usually do not withstand this thermal stress.
`
`[0004]
`Industrially used sterilization with ethylene oxide
`ordinarily requires temperatures of about 40-50° C., typi(cid:173)
`cally for a period totaling about 24-48 hours, inter alia, in
`order to remove the remaining ethylene oxide as completely
`as possible. However, these conditions are often unaccept(cid:173)
`able for temperature-sensitive products. An additional factor
`is that possible residual amounts of ethylene oxide or its
`reaction products in the packaging represent a disadvantage
`of this method because of its carcinogenicity and toxicity.
`
`[0005] Another method, irradiation with y rays or electron
`beams, is also usually unsuitable for sensitive products,
`especially in the liquid stale, because it is associated, for
`example, with losses of activity and/or product degradation.
`
`[0006]
`In the 1980s, a sterilization method which aimed at
`sterilization under reduced pressure ( <1 torr) with hydrogen
`peroxide in the gas phase was described by EP-AO 302 420.
`Although a description of this process at room temperature
`was also given, its sterilization efficiency is inadequate for a
`reliable method for routine use to be developed therefrom.
`The efficiency of this method is increased only by raising the
`temperature to at least 40° C.
`
`[0007]
`In the sterilization method using hydrogen perox(cid:173)
`ide and generation of a gas plasma, chamber temperatures of
`45° C. or 40-45° C. are also usual (T. C. V. Penna, C. A. M.
`Ferraz, and M. A. Cassola; Infection Control and Hospital
`Epidemiology 20: 465-472 (1999) and R. f. Morrissey;
`Biomedical Instrumentation & Technology 30: 404-406
`(1996)). According to U.S. Pat. No. 4,643,876 or EP 207 417
`in fact temperatures of 57° C. arc measured in the material
`to be sterilized, because the method itself leads to warming
`of the sterilization material. Although this method has been
`described as low-temperature method or low-temperature
`plasma, the temperatures which are used or reached are still
`so high that sensitive products are at least partly damaged on
`exposure to this temperature.
`
`[0008] The hydrogen peroxide plasma
`sterilization
`method has been employed, under the name STERRAD®
`method, since the early 1990s in Europe and the USA
`essentially for sterilizing medical instruments. Thus, the
`great majority of the corresponding applications are to be
`found in the hospital sector. There are, however, also a few
`
`applications, for example for products, appliances or dis(cid:173)
`posable articles which can be employed in medicine, where
`the use of ethylene oxide or y irradiation has been prohibited
`for compatibility reasons. These products can then often be
`sterilized without loss of functionality at the temperatures
`which are intrinsic to the hydrogen peroxide plasma steril(cid:173)
`ization method, a chamber temperature of about 45° C.
`
`[0009] The STERRAD® method has
`to date been
`restricted to a chamber temperature of 45° C. (STERRAD®
`100) or >39° C. (STERRAD® GMP 100) because it was
`necessary to assume on the basis of the known results (for
`example, EP-A O 302 420) that effective sterilization can be
`achieved only at temperatures above 39° C. Our own tests
`with the hydrogen peroxide plasma sterilization method at
`the standard chamber temperature of 45° C. with biological
`products under the conditions which are assumed to be mild
`according to the prior art have shown that sensitive biologi(cid:173)
`cal materials in some cases suffered marked or complete
`losses of activity (see example 1). It is thus impossible for
`the latter to be sterilized under the known conditions of the
`STERRAD® method.
`
`DESCRIPTION OF THE INVENTION
`
`[0010] The present invention was based on the object of
`developing a procedure which permits sensitive biological
`and therapeutic products to be sterilized externally in the
`solid or liquid state in their final container (primary pack(cid:173)
`aging). It was moreover intended that the selection of the
`final container ensure that there is no adverse effect on the
`product by the method. 11 was additionally intended for it to
`be possible to sterilize the product in two outer packages
`(secondary packaging).
`
`[0011] This object has been achieved in that it was pos(cid:173)
`sible to develop, on the example of the temperature-sensitive
`components of a fibrin glue, a modification of the hydrogen
`peroxide plasma sterilization method at a further reduced
`temperature which permits final containers with sensitive
`products, even in outer packages, to be efficiently sterilized
`externally in a rapid and mild manner.
`
`BRIEF DESCRIPTION OF THE DRAWINGS
`
`[0012] The scheme depicted shows cartridges as primary
`packaging materials, closed with seal and cap as well as a
`plunger stopper. In order to avoid plunger stopper move(cid:173)
`ments during processes where also vacuum steps are
`applied, the cartridges and plunger stoppers are fixed with a
`spacer within the secondary packaging, e.g. a hard blister
`with Tyvek® lid. In addition to the first secondary packaging
`a further pouch manufactured of a gas permeable material
`may be used.
`
`[0013] A: Cartridge length.
`
`[0014] R: Distance between plunger sloper and car(cid:173)
`tridge end.
`
`[0015] Xl and X2: Length of spacer for fixation of the
`plungers, adjusted in length to the distance between
`plunger and secondary packaging.
`
`DETAILED DESCRIPTION OF THE
`INVENTION
`
`[0016]
`It has now been found that containers with tem(cid:173)
`perature-sensitive products can be effectively sterilized with
`
`Regeneron Exhibit 1018.003
`
`

`

`US 2003/0003014 Al
`
`Jan. 2, 2003
`
`2
`
`hydrogen peroxide/plasma under modified conditions with(cid:173)
`out the previously customary temperatures necessarily being
`used or occurring during this process. At chamber tempera(cid:173)
`tures below 39° C. it is possible to achieve both product
`stability and sterility. At chamber temperatures of 20-39° C.,
`especially also at about 25-35° C., very mild sterilizations of
`products are possible. It is thus possible through introduc(cid:173)
`tion of this low-temperature modification for final containers
`with sensitive products such as, for example, proteins,
`peptides, etc. in solution to he efficiently sterilized. Crucial
`for the low product temperature in this case is both the low
`chamber temperature and the avoidance of an excessive
`energy input, for example on injection of the hydrogen
`peroxide and in the plasma formation.
`
`[0017] Before carrying out the sterilization, the products
`can, where appropriate, be exposed to a preplasma in order
`to remove moisture, as described in EP 707 186, or in order
`to further adapt the product temperature to the chamber
`temperature.
`
`[0018]
`In the sterilization cycle to be applied, it is possible
`for the duration of the so-called injection period(s) and
`diffusion ptriud(s) (whtrt apprupriatt with simultam;uus
`ventilation) to be varied, preferably between about 1 and 60
`minutes. Ventilation during the diffusion period can, where
`appropriate, also be dispensed with, especially if the product
`to be sterilized has a simple geometry.
`
`[0019] The injection of hydrogen peroxide solution can
`also be repeated one or more times--especially if the
`chamber is fully loaded-in order to achieve an adequate
`hydrogen peroxide content in the gas phase and thus an
`increased kill rate. If necessary, it is also possible for the
`tmtirt cyck or parts thtrtuf tu bt repeattd unt or more
`times, although the chamber temperature must be set at <39°
`C. in accordance with the claimed method in order to limit
`the warming.
`
`[0020]
`If the diffusion period with ventilation is omitted,
`it is possible for the injection period, the restoration of an
`adequate vacuum and the plasma to be followed by a half
`cycle of injection, vacuum and plasma. This may with
`simple product geometries reduce the duration of the cycle.
`
`[0021] This method makes it possible for the final con(cid:173)
`tainers of sensitive biological products such as, for example,
`proteins or peptides to be sterilized in short cycles at low
`temperature. As it has heen possible to show in the case of
`blood plasma proteins, a mild sterilization of products in the
`final containers is possible with the method of the invention.
`Such products can then he employed wherever sterile han(cid:173)
`dling is necessary.
`
`[0022] Huwtvtr, it is also pussibk in principle tu apply the
`described procedure to other sensitive biological products
`such as DNA, RNA, lipids, cellular products, etc. No
`significant losses are to be expected with said products
`owing to the short and low temperature stress.
`
`[0023] The method of the invention can, however, also he
`employed generally for sterilizing final containers contain(cid:173)
`ing temperature-sensitive non-biological products. Possible
`examples thereof are synthetic compounds or products
`which can be employed in therapy but which, because of
`their temperature sensitivity, are partly or completely inac(cid:173)
`tivated or damaged in conventional methods.
`
`[0024]
`In the selection of the final container, i.e., the
`primary packaging, care must be taken that movable clo(cid:173)
`sures such as stoppers, plunger seals or caps are fixed so that
`no opening and no leak occurs in the primary packaging
`under vacuum. This can be prevented, for example, by
`appropriate devices, whether by directly fixing the closures
`or hy ensuring hy an appropriate secondary packaging that
`no leak or displacement of stoppers or plunger seals can
`occur (see FIG. 1).
`[0025] A typical half cycle of the method of the invention
`at reduced temperature consists of the following elements:
`[0026] Preparation (to be Carried Out if Required):
`
`[0027]
`lowering the pressure in the treatment cham(cid:173)
`ber, preferably to about 100 to 800 mtorr, very
`preferably to about 300 to 600 mtorr.
`
`[0028] applying a preplasma, preferably for about 1
`to 30 min, very preferably for about 2 to 15 min,
`
`[0029] ventilating, preferably in less than 5 min, very
`preferably in less than 1 min.
`
`[0030] Procedure for a Half Cycle with a Chamber Tem(cid:173)
`perature Set at <39° C. Preferably at About 20-35° C.:
`
`[0031]
`lowering the pressure in the treatment cham(cid:173)
`ber, preferably to about 100 to 800 mtorr, very
`preferably to about 300 to 600 mtorr.
`
`[0032]
`introducing the hydrogen peroxide, usually by
`direct evaporation of a solution in vacuo one or more
`times, with subsequent distribution in the chamber
`(injection), preferably within 20 min, very preferably
`within 15 min,
`
`[0033] where appropriate additional hydrogen perox(cid:173)
`ide diffusion period (where appropriate with venti(cid:173)
`lation of the chamber), depending on the require(cid:173)
`ments for the product to be sterilized, preferably for
`1 to 30 min, very preferably for 3 to 15 min,
`
`[0034]
`renewed lowering of the pressure and resto(cid:173)
`ration of an adequate vacuum,
`
`[0035] generation of a plasma, preferably for 0.5 to
`10 min, very preferably for 1 to 5 min,
`
`[0036] ventilation.
`
`[0037] A complete sterilization cycle usually comprises
`two half cycles. This division has been made for reasons of
`method validation. If the reduction factor achieved in such
`a half cycle with model microorganisms such as, for
`is
`example, spores of Bae. stearothermophilus
`log10
`CFU ~ 6, the method can be said to be efficient with adequate
`certainty.
`[0038] The method to which the invention relates is essen(cid:173)
`tially described in the claims. Examples 2-6 show, on the
`basis of a modified sterilization method for primary pack(cid:173)
`agings containing temperature-sensitive proteins such as, for
`example, the components of a fibrin glue, how effective
`sterilization is possible at low temperature without adversely
`affecting the properties of the protein components.
`[0039] Example 1 shows the result of a hydrogen peroxide
`plasma sterilization run under standard temperature condi-
`
`Regeneron Exhibit 1018.004
`
`

`

`US 2003/0003014 Al
`
`Jan. 2, 2003
`
`3
`
`tions according to the prior art. The following examples
`(2-6) are intended to illustrate the principle of the modified
`method at low temperature.
`
`COMPARATIVE EXAMPLE 1
`[0040] Various protein solutions (main constituent of pro(cid:173)
`tein solution 1: fibrinogen, of protein solution 2: factor XIII
`and of protein solution 3: thrombin) were dispensed into
`glass carpules and sealed into bags consisting of Tyvek sheet
`and a transparrnt plastic sheet. The bags were then put in
`layers into baskets and treated in a STERRAD® GMP 100
`sterilizer (supplier: Johnson & Johnson Medical GmbH,
`22844 Norderstedt, Germany) with a hydrogen peroxide
`plasma sterilization method in accordance with the follow(cid:173)
`ing parameters. The exact composition of the protein solu(cid:173)
`tions used for carrying out the examples is immaterial. The
`solutions are ones known per se to the skilled worker and
`comprise biological proteins of natural or recombinant ori(cid:173)
`gin employed in therapy.
`[0041] Preparation:
`[0042] Chamber temperature set at 45° C. Loading of the
`chamber: basket charged with product-containing primary
`packagings in bags.
`
`[0043] Procedure for the 1st Half Cycle:
`[0044] Vacuum: to about 400 mtorr
`[0045]
`Injection: about 6 min (1800 µ1 of 59% H 20 2 )
`[0046] Diffusion: 10 min (including ventilation)
`[0047] Vacuum: to about 400-500 mtorr
`[0048] Plasma: 2 min
`[0049] A 2nd, 3rd and 4th half cycle ( corresponding
`to the 1st half cycle) are carried out directly follow(cid:173)
`ing half cycle 1.
`[0050] Results of the sterilization runs in example 1:
`stability of the investigated protein solutions:
`
`Content (% of initial levels)
`
`Protein solution
`1:
`Fibrinogen
`
`Protein
`solution 2:
`Factor XIII
`
`Protein
`solution 3:
`Thrombin
`
`100
`
`Gel formulations;
`material unusable
`100
`
`100
`
`100
`
`100
`
`100
`
`26.0
`(turbidity)
`100
`
`Stage
`
`Content before
`sterilization
`Content after 4
`half cycles
`Untreated
`control
`
`[0051] As is evident from the table for example 1, a
`temperature-dependent aggregation or degradation or dena(cid:173)
`turation occurs in the case of protein solution 1 ( containing
`fibrinogen) and protein solution 3 (containing thrombin).
`This means that both products have become unusable for the
`intended application through the hydrogen peroxide plasma
`sterilization method according to the prior art.
`
`EXAMPLE 2
`[0052] Various protein solutions (main constituent of pro(cid:173)
`tein solution 1: fibrinogen, of protein solution 2: factor XIII
`
`and of protein solution 3: thrombin) were dispensed into
`glass carpules and sealed into bags consisting ofTyvek sheet
`and a transparent plastic sheet. The bags were then put in
`layers into baskets and treated in a STERRAD® GMP 100
`sterilizer with a hydrogen peroxide plasma sterilization
`method in accordance with the following parameters.
`
`[0053] Preparation:
`
`[0054] Chamber temperature set at: 30° C.
`
`[0055] T mding of the chamber: lower basket tightly
`packed with the described bags containing water(cid:173)
`filled primary packaging; upper basket charged with
`product-containing hags and, in addition, a tempera(cid:173)
`ture sensor.
`
`[0056] Loading Followed by Carrying Out a "Preplasma":
`
`Vacuum:
`Prcplasma:
`Ventilation
`
`to about 400 mtorr
`5 min
`(brief)
`
`[0057] Procedure for a 1st Half Cycle:
`
`Vacuum:
`Injection:
`Diffusion:
`Vacuum:
`Plasma:
`
`to about 400 mtorr
`about 12 min (1 800 µl of 59% H20 2 )
`5 min (including ventilation)
`to ahout 400-500 mtorr
`about 2 min
`
`[0058] Vacuum (brief) and ventilation (to remove
`sample)
`
`[0059] A 2nd half cycle ( corresponding to the 1st half
`cycle) is carried out directly following half cycle 1.
`
`[0060] Results of the sterilization runs in example 2:
`stability of the investigated protein solutions:
`
`Content (% of initial levels)
`
`Protein solution
`1:
`Fibrinogen
`
`Protein
`solution 2:
`Factor XIII
`
`Protein
`solution 3:
`Thrombin
`
`100
`
`107.7
`
`104.3
`
`100.9
`
`100
`
`101.4
`
`100.5
`
`99.5
`
`lUU
`
`98.5
`
`98.2
`
`98.6
`
`Stage
`
`Content before
`sterilization
`Content after
`1st half cycles
`Content a'rter 2
`half cycles
`Untreated
`control
`
`EXAMPLE 3
`[0061] Various protein solutions (main constituent of pro(cid:173)
`tein solution 1: fibrinogen, of protein solution 2: factor XIII
`and of protein solution 3: thrombin) were dispensed into
`glass carpules and sealed into bags consisting ofTyvek sheet
`and a transparent plastic sheet. The bags were then put in
`layers into baskets and treated in a STERRAD® GMP 100
`
`Regeneron Exhibit 1018.005
`
`

`

`US 2003/0003014 Al
`
`Jan. 2, 2003
`
`4
`
`sterilizer with a hydrogen peroxide plasma sterilization
`method in accordance with the following parameters.
`
`[0062] Preparation:
`
`[0063] Chambt:r kmperature st:l at: 30° C.
`
`[0064] Loading of the chamber: lower basket tightly
`packed with the described bags containing water(cid:173)
`filled primary packaging; upper basket charged with
`product-containing bags and, in addition, a tempera(cid:173)
`ture sensor.
`
`[0065] T nading Followed hy Carrying Out a "Preplasma":
`
`[0066] Vacuum: to about 400-500 mtorr
`
`[0067] Preplasma: 5 min
`
`[0068] Ventilation (brief)
`
`[0069] Procedure for a 1st Half Cycle:
`
`[0070] Vacuum: to about 400 mtorr
`
`[0079] Preparation:
`[0080] Chamber temperature set at: 35° C.
`[0081] Loading of the chamber: lower basket tightly
`packed with the described bags containing water(cid:173)
`filled primary packaging; upper basket charged with
`product-containing bags, bags containing spore
`strips and primary packaging, and additionally a
`temperature sensor.
`[0082] After loading a "preplasma" is carried out:
`
`Vacuum:
`Preplasma:
`Ventilation
`
`to about 400-500 mtorr
`5 min
`(brief)
`
`[0083] Procedure for a 1st Half Cycle:
`
`[0071]
`Injection: about 17 min (1 800 µl of 59%
`H202)
`
`[0072] Vacuum: to about 400-500 mtorr
`
`Vacuum:
`Injection:
`Diffusion:
`Vacuum:
`Plasma:
`
`lo about 400 mlorr
`about 12 min (1 800 µl of 59% H202)
`5 min (including ventilation)
`to about 400-500 mtorr
`2min
`
`[0073] Plasma: 2 min
`
`[0074] Vacuum (brief) and ventilation (to remove
`sample)
`
`[0075] A 2nd half cycle (corresponding to the 1st half
`cycle) is carrit:d out directly following half cycle 1.
`
`[0076] Results of the sterilization runs in example 3:
`stability of the investigated protein solutions:
`
`[0084] Vacuum (brief) and ventilation ( to remove
`sample)
`[0085] A 2nd half cycle ( corresponding to the 1st half
`cycle) is carried out directly after half cycle 1.
`[0086] Results of the sterilization runs in example 4:
`stability of the investigated protein solutions:
`
`Content(% of initial levels)
`
`Protein solution
`1:
`Fibrinogen
`
`Protein
`solution 2:
`Factor XIII
`
`Protein
`solution 3:
`Thrombin
`
`100
`
`101.7
`
`101.2
`
`100.9
`
`100
`
`101.9
`
`101.9
`
`99.5
`
`10'1
`
`98.9
`
`98.6
`
`98.G
`
`Stage
`
`Content hefore
`sterilization
`Content after
`1st half cycles
`Content after 2
`half cycles
`Untreated
`control
`
`EXAMPLE 4
`[0077] Various protein solutions (main constituent of pro(cid:173)
`tein solution 1: fibrinogen, of protein solution 2: factor XIII
`and of protein solution 3: thrombin) were dispensed into
`glass carpules and sealed into bags consisting of Tyvek sheet
`and a transparent plastic sheet. The hags were then put in
`layers into baskets and treated in a STERRAD® GMP 100
`sterilizer with a hydrogen peroxide plasma sterilization
`method in accordance with the following parameters.
`[0078]
`In addition to product-filled primary packagings, in
`order to check the efficiency of sterilization carpules and
`strips with spores of a test organism (Bae. stearothermophi(cid:173)
`lus) were sealed doubly in Tyvek bags.
`
`Content (% of initial levels)
`
`Protein solution
`1:
`Fibrinogen
`
`Protein
`solution 2:
`Factor XIII
`
`Protein
`solution 3:
`Thrombin
`
`100
`
`106.9
`
`102.7
`
`100.9
`
`100
`
`108.0
`
`104.7
`
`99.5
`
`100
`
`99.3
`
`99.6
`
`9R6
`
`Stage
`
`Content before
`sterilization
`Content after
`1st half cycles
`Content after 2
`half cycles
`Untreated
`control
`
`[0087] Sterility assessment of 8 spore strips treated in a
`half cycle:
`
`Sterility assessment of the employed spore strips No.
`
`2
`
`3
`
`4
`
`5
`
`7
`
`8
`
`2.2
`
`2.2
`
`2.2
`
`2.2
`
`2.2
`
`2.2
`
`2.2
`
`2.2
`
`0
`
`0
`
`0
`
`0
`
`0
`
`Kumber of
`spores per
`spore strip
`before the half
`cycle ( x 106
`)
`Content after
`1st half cycles
`
`Regeneron Exhibit 1018.006
`
`

`

`US 2003/0003014 Al
`
`Jan. 2, 2003
`
`5
`
`EXAMPLE 5
`
`[0088] Various protein solutions (main constituent of pro(cid:173)
`tein solution 1: fibrinogen, of protein solution 2: factor XIII
`and of protein solution 3: thrombin) were dispensed into
`glass carpules and sealed into bags consisting of Tyvek sheet
`and a transparent plastic sheet. The bags were then put in
`layers into baskets and treated in a STERRAD® GMP 100
`sterilizer with a hydrogen peroxide plasma sterilization
`method in accordance with the following parameters.
`
`[0089] Preparation:
`[0090] Chamber temperature set at: 35° C.
`[0091] Loading of the chamber: lower basket tightly
`packed with the described bags containing water(cid:173)
`filled primary packaging; upper basket charged with
`product-containing bags and, in addition, a tempera(cid:173)
`ture sensor.
`[0092] Loading Followed by Carrying Out a "Preplasma":
`
`Vacuum:
`Preplasma:
`Ventilation
`
`to about 400-500 mtorr
`5 min
`(brief)
`
`[0093] Procedure for a 1st Half Cycle:
`
`Vacuum:
`1st injection:
`2nd injection:
`Diffusion:
`Vacuum:
`Plasma:
`
`to about 400 mtorr
`about 2 min (1 800 µl of 59% H 2 0 2)
`about 10 min (1 800 µl of 59% H 70 7 )
`5 min (including ventilation)
`to about 400-500 mtorr
`2 min
`
`[0094] Vacuum (brief) and start with next half cycle
`or ventilation (to remove sample)
`
`[0095] A 2nd, 3rd and 4th half cycle (corresponding
`to the I st half cycle) are carried out directly follow(cid:173)
`ing half cycle 1.
`[0096] Results of the sterilization runs in example 5:
`stability of the investigated protein solutions:
`
`Content(% of initial levels)
`
`Protein solution
`1:
`Fibrinogen
`
`Protein
`solution 2:
`Factor XIII
`
`Protein
`solution 3:
`Thrombin
`
`100
`
`94.5
`
`92.0
`
`97.5
`
`100
`
`102.9
`
`108.1
`
`106.2
`
`100
`
`100.3
`
`100.3
`
`99.3
`
`Stage
`
`Content before
`sterilization
`Content after 2
`half cycles
`Content after 4
`half cycles
`Untreated
`control
`
`materials, water-filled glass carpules were sealed into outer
`packages consisting of PET hard blisters, closed with Tyvek
`paper, and additionally with bags consisting of Tvvek paper
`and a transparent plastic sheet. The packages were than put
`in tight layers into baskets and treated in a STERRAD®
`GMP 100 sterilizer with a hydrogen peroxide plasma ster(cid:173)
`ilization method according to the following parameters.
`
`[0098] Strips of spores of a test organism (Bae. stearo(cid:173)
`thermophilus) were introduced into the double packaging
`and sealed in the least accessible positions of eight blister
`packs distributed at different positions in the baskets in order
`to check the sterilization efficiency.
`
`[0099] Preparation:
`
`[0100] Chamber temperature set at: 32° C.
`
`[0101] Loading of the chamber: lower and upper
`basket tightly packed with the described packages
`(172 items) containing water-filled primary packag(cid:173)
`ing; 8 packages contained in addition to the primary
`packagings also spore strips (between carpule and
`hard blister or between carpule end and stopper
`retainer) and were distributed in accordance with a
`defined plan in the baskets.
`
`[0102] Procedure for a "Preplasma":
`
`Vacuum:
`Preplasma:
`
`to about 400-500 mtorr
`5 min
`
`[0103] Procedure for a Half Cycle:
`
`Vacuum:
`3 injections:
`
`Diffusion:
`Vacuum:
`Plasma:
`
`to about 400-500 mtorr
`lasting ahout 3, 6 and 6 min respectively
`(each of 1 800 µl of 59% Hpzl
`5 min (including ventilation)
`to about 400-500 mtorr
`2 min
`
`[0104] Vacuum (brief) and ventilation ( to remove
`sample)
`
`[0105] Results of the sterilization run in example 6: ste(cid:173)
`rility assessment of 8 spore strips treated in a half cycle:
`
`[0106] All of the spore strips employed were completely
`inactivated, even those in the least accessible positions, i.e.,
`this method made it possible to sterilize more than 106
`spores effectively in one half cycle.
`
`EXAMPLE 6
`
`[0097] To test the sterilization efficiency of the method in
`a maximally loaded chamber with commercial packaging
`
`Kumber of
`sp01es per
`spore strip
`before the half
`cycle ( x 106 J
`
`Sterility assessment of the employed
`s ore stri s Ko.
`
`2
`
`3
`
`4
`
`5
`
`7
`
`8
`
`26
`
`26
`
`26
`
`26
`
`26
`
`2~
`
`26
`
`26
`
`Regeneron Exhibit 1018.007
`
`

`

`US 2003/0003014 Al
`
`Jan. 2, 2003
`
`6
`
`-continued
`
`Sterility assessment of the employed
`s ore stri s No.
`
`2
`
`0
`
`4
`
`5
`
`0
`
`0
`
`7
`
`Spores de(cid:173)
`tectable after a
`half cycle
`
`0
`
`1. A process for hydrogen peroxide plasma sterilization
`comprising:
`
`(a) inserting at least one primary container containing a
`temperature-sensitive material into a sterilization treat(cid:173)
`ment chamber;
`
`(b) lowering the pressure in the treatment chamber to
`create a vacuum;
`
`( c) injecting, at least one time, hydrogen peroxide into the
`chamber;
`
`( d) lowering the pressure in the treatment chamber to
`reestablish a vacuum;
`
`( e) generating a plasma; and
`
`(t) ventilating the chamber;
`
`wherein the chamber temperature is less than 39° C.
`throughout the process.
`2. The process of claim 1, wherein the pressure in step (b)
`is about 100 to 800 mtorr.
`3. The process of claim 1, wherein step (c) is performed
`from between 1 and 60 minutes.
`4. The process of claim 1, wherein prior to step (d) a
`hydrogen peroxide diffusion step is performed simulta(cid:173)
`neously with ventilation.
`5. The process of claim 1, wherein prior to step (d) a
`hydrogen peroxide diffusion step is performed without ven(cid:173)
`tilation.
`6. The process of claim 4, wherein the hydrogen peroxide
`diffusion step is performed from between i and 60 minutes.
`7. The process of claim 5, wherein the hydrogen peroxide
`diffusion step is performed from between 1 and 60 minutes.
`
`8. The process of claim 1, wherein the temperature of the
`temperature-sensitive material does not rise above 40° C.
`during the sterilization process.
`9. The process of claim 1, wherein the temperature(cid:173)
`sensitive material comprises biological materials.
`10. The process of claim 9, wherein the biological mate(cid:173)
`rials are proteins, peptides, nucleic acids, lipids, or cellular
`materials.
`11. The process of claim 9, wherein the biological mate(cid:173)
`rial is a fibrogen containing solution.
`12. The process of claim 9, wherein the biological mate(cid:173)
`rial is a Factor XIII containing solution.
`13. The process of claim 9, wherein the biological mate(cid:173)
`rial is a thrombin containing solution.
`14. The process of claim 9, wherein the biological mate(cid:173)
`rial comprises the components of tissue glue.
`15. The process of claim 9, wherein the biological mate(cid:173)
`rial comprises the components of fibrin glue.
`16. The process of claim 1, wherein the temperature(cid:173)
`sensitive material comprises non-biological materials.
`17. TI1e process of claim 1, wherein the process is
`performed more than one time.
`18. The process of claim 1, wherein before step (b ), a
`preplasma step is performed comprising:
`
`lowering the pressure in the treatment chamber to create
`a vacuum;
`
`applying a preplasma; and
`
`ventilating the chamber.
`19. The process of claim 18, wherein the pressure is about
`100 to 800 mtorr.
`20. The process of claim 18, wherein the preplasma is
`applied for about 1 to 30 minutes.
`21. The process of claim 18, wherein the ventilation step
`is no greater than 5 minutes.
`22. The process of claim 1, wherein the primary container
`is enveloped at least one time with materials partially
`permeable to hydrogen peroxide.
`23. The process of claim 1, wherein the primary container
`containing the temperature-sensitive material is placed in a
`secondary container.
`
`* * * * *
`
`Regeneron Exhibit 1018.008
`
`