JP2601551C - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION   The present invention relates generally to viral inactivation methods, and in particular, to biologically active therapeutic proteins.
For virus inactivation in cytoplasmic products.   In summary, the present invention is directed to purified biologically purified cells obtained from blood or cell culture systems.
Lipid-enveloped viruses present in active protein products
) May cause the protein product to sediment adversely or
Sufficient non-ion to inactivate the virus without adversely affecting activity
Contact the product with caprylic acid in caprylic acid concentration, pH, temperature and ionic environment
By doing so, it can be inactivated.   Prior art   The safety of a pharmaceutical product is particularly important where viral contamination can occur (e.g.,
From blood, or from cell culture systems designed to produce biologically active proteins
Is always a concern. Unfortunately, virus found
The product itself is compatible with many known and conventional virus inactivation techniques.
It is often unstable and very susceptible. In some cases, tampa
Attempting to protect the virus also protects the virus.   Various attempts have been made to overcome this situation. For example, some control
Heat treatment or specifically selected chemicals to remove biologically active proteins.
It is well known that it can be activated. Adverse effects on protein biological activity
Inactivate the virus without causing or reducing its amount to a problem.
To do this, several heat treatment methods have been developed. For example, Fernandes
U.S. Pat. No. 4,440,679 (ndez) and Lundblad (Factor VIII
Carbohydrate stabilization for pasteurization of very unstable coagulation proteins known as
U.S. Pat. No. 4,762,714 to Mitra and Mozen (Wigs in immunoglobulin products under controlled conditions of pH, temperature and time)
(Indicating Luz inactivation). Prisoner VIII is first freeze-dried, then pasteurized (
(At least 10 hours at 60 ° C).
U.S. Pat. No. 4,456,590 to Reubenstein and Thomas
mas), US Pat. No. 4,495,278 (similar heat treatment of lyophilized Factor IX).
I want to be illuminated.   Various chemical methods have also been used to inactivate the virus. For example, Lemba
U.S. Pat. No. 4,534,972 to Lenbach (copper phenanthroline and related patents).
US Patent No. 4,481,189 to Horowitz (US Patent No. 4,481,189).
Use of tri-n-butyl phosphate and related compounds).   Production of plasma products (globulin precipitation) and lipid-coated virus
Even carboxylic acids such as caprylic acid were used for inactivation of iruses)
Has not been used in the presence of therapeutic biologically active proteins [J
-An anti-micro vial agent such as J.A.Sands
De Hemotherapy (Anti Microbial Agents and Chemotherapy), January 1979
134-136]. Carboxylic acid (sodium caprylate) is a factor VIII
It has also been used as a combination with heat and amino acids for irus inactivation.
No. 4,446,134 to Ito et al.). Horowitz etc.
Plasma induction disclosed by Vox Sang, 54: 14-20 (1988).
See also, Sequential use of fatty acids for virus inactivation of conductors.   Caprylic acid without affecting IgG, ceruloplasmin and IgA Precipitation of plasma proteins by the method described by Steinbach et al.
Steinbuch, M. and Audran, R., Archives of
・ Biochemistry and Biophysics (Arch. Biochem. Biophys)
), 134, 179-294 (1969)]. Human, horse, sheep and rabbit blood
Diluting the serum or plasma with 0.06 M acetate buffer to about 1.7% protein;
Adjusted to pH 4.8 at 20 ° C., 0.174N (2.5% by weight) with respect to caprylic acid
And Note the buffer molarity (0.06M) and pH (pH 4.8 ± 0.05)
This is important for high purity IgG.   The precipitation method of Steinbach-M, supra, describes a method for recovering IgG from 0.0
Using caprylic acid at a concentration of 66M (0.86% by weight), hybridoma culture waste
Applied to medium and ascites fluid from mice [Lusso, C., Carregaro, L.
Nza E, Feron S; Journal of Immunological Methods,
65, 269-271 (1983) (Russo, C., Callegaro, L., Lanza, E., Ferron
e, S., J. Immunol. Methods, 65, 269-271 (1983)]. The same method is used for caprylic acid 0
. Applied to diluted human plasma adjusted to 15M or 2.16% by weight [Habibe
Habeeb, A.F.S.A.an, A.F.S.A. and Francis, E.R.
d Francis, E.R). , Preparative Biochemistry (Prep.Biochem),
14 (1), 1-17 (1984)]. DEAE cellulose adsorption and elution
Isolated from Cohn cold ethanol Fraction III
A is further purified by caprylic acid precipitation to remove α-2 macroglobulin
[Pejaudier, L .; , Audran, Earl
n, R) and Stein Back, M (Steinbuch, M). , Vox Sang, 23, 16
5-175 (1972)]. The parameters for sedimentation were 0.9% sodium chloride
2.0% protein concentration, pH5 and 0.078M or 1.12
It consisted of caprylic acid added at room temperature to give a percentage by weight. Sinked α
-2 macroglobulin was removed by centrifugation.   Most of the non-immunoglobulin present in the cold ethanol fraction III of corn
Caprylic acid has been used to precipitate proteins and lipoproteins [
Inback, M. Audran, Earl. , Pejodir, El. , Bratri
Box, C (Blatrix, C). , Preparative Biochemistry, 3 (4)
363-373 (1973)]. About 2.5% protein of the suspension of fraction III
The pH was adjusted to 0.05 N acetate at 4.8 and allowed to come to room temperature.
Caprylic acid was added to make 0.174M or 2.5% by weight. Discard the resulting sediment
I was The supernatant was rich in IgG, IgM and IgA. Capryl as a settling agent
In all cases where acids are used, ideal conditions of pH, temperature and effectiveness of caprylic acid
Note that caprylic acid is present in significant amounts above its maximum solubility in water under
It should be. Such amounts, about 0.86-2.5% by weight, are usually insoluble
Sufficient amount of relatively insoluble caprylic acid in the form (emulsion) and thus precipitation
Necessary to guarantee useful amounts of the agent.   Despite the numerous publications mentioned above, the biological activity or recovery of therapeutic proteins
Inactivates virtually all lipid-coated viruses without adversely affecting yields
At a concentration lower than the sedimentation concentration and alone There is no known method of using caprylic acid. Carefully control operating conditions
A biologically active therapeutic production without adversely affecting the activity of the product
That caprylic acid can be used to inactivate lipid-coated viruses in
We found them. The details of our invention are described below.   The advantage of our method is that non-ionized caprylic acid generated according to the following dissociation reaction
The amount is based on precise control.   CH_Three(CH_Two)₆COOH CH_Three(CH_Two)₆COOH^-+ H⁺   Caprylic acid ionized form   Thus, substantially all fats in unstable, biologically active therapeutic products
The method of the present invention for inactivating dendritic coat viruses involves reducing the amount of non-ionized caprylic acid.
Control and have no adverse effect on the amount of product, biological activity and therapeutic efficacy.
Under sufficient concentration, pH and ionic environment to ensure virus inactivation
Contacting the product with caprylic acid.   In a preferred embodiment, the inactivation method of the present invention comprises the specific tank to be inactivated.
Controls the amount of caprylic acid generated without adversely affecting the stability or amount of protein
The selective use of caprylate concentration and pH is contemplated. 0.068%
More or less than the maximum solubility of about 0.07% of caprylic acid.
Ensuring the case avoids unwanted loss of protein. About 0.
Caprylic acid concentration exceeding 07% (weight% basis) is
May cause formation of undesired proteins, resulting in undesirable protein loss.
Sir. On the other hand, the amount of caprylic acid can inactivate lipid-coated viruses within a reasonable time. Be sufficient (at least 0.001%). Amazing
In particular, at lower pH (6.5 or lower), caprylic acid is 0.07-0.
. If present at 001%, virus inactivation is achieved almost immediately.
At higher pH, if caprylate is clearly the main species, the virus
Higher concentrations and longer times are required to achieve the same log reduction of Scratch
Thus, in a preferred embodiment, the caprylic acid concentration useful in the present invention is about 0% by weight.
. 07 to about 0.001%.   A first advantage of the present invention is its flexibility. At low concentrations, the invention is used at low pH
Thus, some viruses can be immediately inactivated. This method is necessary
Inactivated at higher pH, at higher concentrations and for longer periods of time
It is also possible. This is because the protein in question is altered during virus inactivation or
Choose inactivation at the most stable pH of the protein in question so that it is not destroyed
Is also possible. Thus, if preferred, the caprylic acid concentration is in weight / weight%
In water, it is in the range of about 0.07% to about 0.001% in water, which is pH and
As described in more detail below, in the ionized form (i.e., such as sodium caprylate).
Control) by controlling both the amount of caprylate in Lucrative
In short, caprylic acid has low, harmless toxicity to humans and
A stabilizer for albumin or plasma protein fraction (PPF) which is perfused in large quantities to the patient;
And is currently being used. Another advantage of the present invention is that
Contains viruses such as IgM / IgG purified from certain corn fraction III pastes.
To ensure that a therapeutically active protein is available. Basic profit
The point is that this method With careful control, it is gentle on proteins and has a pH of about 4.0
A method that can be applied to any protein that is stable at pH 8.0
It is.   By the standard biochemical convention and as used herein, the subscript "ate" ("a
te ”) (ie, caprylate) is the acid and its ionized form, as in the dissociation reaction.
It represents a mixture with the state.   CH_Three(CH_Two)₆-COOH CH_Three(CH_Two)₆-COO^-+ H⁺   Caprylic acid ionized form The pKa of caprylic acid is 4.89 [CRC Handbook of Chemistry and Physics (CRC
 HandbooK of Chemistry and Physics), 56th Edition] Henderson-Hasselbal
Henderson-Hasselbalch equation: Gives the concentration of the acid and its ionized form at various pHs. Thus, pH and cap
Careful control of the relate concentration facilitates the desired concentration of caprylic acid.
Obtainable. For example, keep the caprylic acid concentration at 0.07% (0.0035M).
And the ionized form (eg, sodium caprylate) has a pH of 4.9.
If it changes between 0.06% and 2.0% at pH = 8, consider FIG.
The amount of caprylic acid indicated is produced. As used herein, caprylic acid is
Refers to the non-ionized form of the acid (also known as octanoic acid).   It is relatively easy to achieve a practical and favorable concentration of caprylic acid.
We have found. This is 0.1% at pH 4.8 and 2 at about pH = 9.
By changing the concentration of sodium caprylate between 0% and
Done by causing immediate inactivation Can be More preferably, the total concentration of caprylate is 0.1% at pH 4.8.
Immediate virus keeping linearly increasing between 2.0 and 2.0% at 6.5
Causes inactivation. Alternatively, caprylate may be used for a longer period of time (eg,
4 hours) Keep the pH between 6.5 and 9.5 at 2% to provide a suitable cap for virus inactivation.
A lylic acid concentration can be provided. Anyway, non-ionized acid form (caprylic acid)
By acting on the lipid envelope or the protein embedded therein,
Caprylic acid is converted to an ionized form by its activator of killing irs and its dissociation reaction.
Virus at high pH by increasing the concentration of sodium (caprylate)
The premise is that concentrations can be kept high enough to kill. Protein composition and wi
The mixture with the ruth and the bacterial inactivator is usually at least about 0.25 hours (
(Preferably about 0.5-3 hours) for about 2-60 ° C (preferably about 4-20 ° C).
Kept at temperature. As mentioned above, the process of the present invention comprises a protein material to be treated.
It is usually performed under pH conditions that are compatible with the material. Thus, protein dependent
Thus, the pH of the mixture is generally about 4-1 for most biologically active proteins.
0, preferably about 4.5-8.5, and more preferably about 4.8-8.0.
Should be. Generally, the pH and temperature range will be relative to the active protein in the composition.
Selected to ensure that the minimum obstacle is minimal. Those skilled in the art can
We know the preferred pH range for quality.   The person skilled in the art also adds suitable stabilizers for the protein in question during the processing of the aforementioned mixtures.
Will. The protein composition is then processed and added to caprylic acid / cap
The relate can be removed. Conventional methods can be used to achieve this purpose.
Wear. For example, the mixture can be dialyzed Alternatively, the protein of interest is bound to an anion exchange resin and washed to remove the protein of interest.
Removal of added caprylic acid / caprylate by subsequent elution of the protein
Can be. Other means for removing this agent will occur to those skilled in the art.
.   We have determined that our virus inactivation treatment is antibody (plasma-derived and monoclonal).
), Human serum alpha albumin, coagulation factor, fibronectin and transfer
To be effective against a wide range of biologically active therapeutic proteins such as
I found it.   Definition of terms   Caprylic acid means the non-ionized form as described above. As used herein
A substantial decrease in infectivity is defined as a virus infectivity titer of a given formulation of at least 4
Or to an undetectable (ND) level. Virtually immediate
Virus inactivation means that the virus titer can be measured using a rapid routine method
Virus inactivation occurs (eg, it is ND (not detectable))
Means that. Capryl does not adversely affect biological activity
Contact with the acid is carried out using conventional methods for the biologically active protein in question.
Means less than about 30% loss of the original biological activity as measured by
. Not adversely affecting the amount of biologically active protein
The contact with the lylic acid is about 40% of the amount of the original protein (the amount before the virus inactivation treatment).
Means less than% loss. This is preferably less than about 10%.
Good. As used herein, a lipid-encapsulated virus is a virus whose nucleic acid contains lipids.
A virus that is encapsulated by a pseudoside. These are for those skilled in the art
Well known and lipid coatings The expression enveloping (or coating) virus is also well known. Therapeutic means administered to mammals
Means that it can have a medically beneficial effect when performed.   Variables such as concentration of caprylic acid and caprylate, pH, temperature and time are viruses
How infectivity decreases, protein recovery and retention of protein biological activity
An example of the disclosure of the present invention is provided below to show whether it can be affected.   Example 1   Epstein-Barr virus (EBV) transformed human B
Lymphocytes (ATCCC CRL8752) cause Pseudomonas aerug
inosa) monoclonal IgM antibody (human) (PsMAb-IgM) was produced. This in advance
The transformed cells were stained with seven Fisher-Devlin serotypes (Fisher-Devlins serotypes).
Donation with naturally occurring antibody titer specific to F-4, one of the
Obtained from This antibody binds to a serotypic determinant on the bacterial surface lipopolysaccharide.
Join. The cell culture harvest for the F-4 antibody is clarified, partially purified and
The pH was adjusted to 8 with 1N NaOH. 0.38 mg / ml at room temperature (RT)
IgM solution (total 19 mg) was first added to Herpes Simplex Form I (Herpes S).
implex type I) (HSV-1) virus and vesicular stomatitis virus (vesicular st
omatitis virus) (VSV). 2.0% by weight of sodium caprylate
Lithium was added and the solution was readjusted to pH8. This is the Henderso at pH8
0.0014% by weight of caprylic acid and ionized form
Equivalent to 1.986% by weight of the form of caprylate. Virus spiking
 spiking) The solution was held for 60 minutes. The virus-spiked pretreatment sample is
It was used as a control in the examples and later examples.   The PsMAb-IgM F-4 protein yield value was determined by radioimmunodiffusion (RID) assay.
Was. Functional activity was determined as specific ribopolysaccharide (LPS) binding capacity. afterwards
Studies of the IgMF-4 example showed substantially similar LPS binding capacity.   Table 1 shows the results of Example 1.   Example 2   PsMAb-IgM (F-4) was prepared and, in addition to VSV,
Virus virus (EBV) and the inactivation step at a lower temperature (5 ° C).
Except for this, processing was performed in the same manner as in Example 1. Spiking with virus
After that, the solution was held for 120 minutes.   The results of Example 2 are shown in Table II.   Example 3   The partially purified IgM solution for inactivation was at 5 ° C. and 1N HCl
Was adjusted to pH 4.8 by the same procedure as in Example 1 except that the pH was adjusted to 4.8.
IgM (F-4) was prepared. This sample was first contacted with the virus and then 0.1%
Weight percent caprylic acid was added and readjusted to pH 4.8 with 1N NaOH. This
It is calculated from Henderson-Hasselbarf calculation at pH 4.8, which is caprylic acid.
Corresponds to 0.055% by weight and 0.045% by weight of caprylate in ionized form
. After virus spiking, the solution was kept at 4 ° C. for 60 minutes.   The results of Example 3 are shown in Table III.   Example 4   Obtained from donors with naturally occurring antibody titers to Pseudomonas aeruginosa exotoxin A
EBV-transformed human B lymphocytes (ATCCC CRL8833;
Domonas exotoxin A antibody IgG (human) (MAb ExoA IgG) was produced. MAb
  The cell culture harvest of ExoA IgG was clarified and purified. MAb at 5 ° C
ExoA IgG was adjusted to 0.5 mg / ml (total 25 mg) at pH 6.5 and V
Contact was made with the SV and HSV-1 viruses. 2.0% by weight of caprylic acid
Sodium was added and the pH was readjusted to 6.5 with 1N HCl. This is pH6
. Caprylic acid 0.042 according to Henderson-Hasselbalf calculations in Example 5
% By weight and 1.958% by weight of caprylate in ionized form. Spike
After aging, the solution was held for 30 minutes.   The results of Example 4 are shown in Table IV.   Example 5   As shown in the results, two concentrations of sodium caprylate were used.
Except for the above, MAb ExoA IgG was prepared in the same manner as in Example 4. Will
The sample for the study of the inactivation efficiency was pH 6.3 and indirect at 5 ° C. for 60 minutes.
Touched. Virus species include VSV, HSV-1, vaccinia virus and Sind
Bisviruses are included.   Table 5 shows the results of Example 5.   The A * sample received 1.0% by weight of sodium caprylate and 1N HCl
PH was adjusted again to 6.3. This is Henderson Hassel at pH 6.3
Caprylic acid 0.033% by weight and cap ion in ionized form according to Balff's calculation
0.967% by weight.   B ** sample received 2.0% by weight of sodium caprylate and 1 NHC
pH was readjusted to 6.3. This is Henderson-Hasse at pH 6.3.
0.065% by weight of caprylic acid and capryle in ionized form
Corresponding to 1.935% by weight.   Example 6   Two non-lipid coated viruses, Bovin Parvovirus (B)
PV) and polio II with caprylic acid to inactivate lipid-coated viruses
In contrast, it proved ineffective. Ps MoAb Ig M (F-4) was prepared and the inactivation temperature was 5 ° C. and the virus was BPV
Chemically treated as in Example 1, except that IgG samples
First contact with BPV virus and polio II virus, then nato caprylate
Lithium was added to 2.0% by weight and readjusted to pH 6.3 with 1N HCl.
Was. This is calculated by Henderson-Hasselbalf calculations at pH 6.3.
0.065% by weight of lylic acid and 1.935% by weight of caprylate in ionized form
Hit. After virus spiking, the solution was held for 120 minutes.   The results of Example 6 are shown in Table VI.   Example 7   From the supernatant IV-4 of the cold ethanol purification method of E.J.
Serum albumin was isolated. 0.5 mg / ml of this albumin at pH 6.0
(Total 25 mg). At 4 ° C, add this albumin solution to vesicular stomatitis
Rus (VSV), then 1.0% by weight of sodium caprylate
And the pH was readjusted to 6.0 with 1N HCl. This is the hen at pH6
0.062% by weight of caprylic acid and ions as calculated by Darson-Hasselbarf
Capryle Corresponding to 0.938% by weight. After virus spiking, remove the solution for 60 minutes
Held.   The results of Example 7 are shown in Table VII.  Example 8   Human serum proteins rich in unstable coagulation factors II, VII, IX and X
By anion exchange adsorption from effluent I of the cold ethanol purification method of J. Corn
Prepared. The eluted coagulation factor was further purified and at pH 6.8 1.73 mg /
The protein concentration was adjusted to ml. 2.0% by weight sodium caprylate
And readjusted to pH 6.8 with 1N HCl. This is pH 6.8
0.022 weight of caprylic acid according to Henderson-Hasselbalf calculations
% And 1.978% by weight of caprylate in ionized form. 2 hours at 4 ° C
Coagulation protein after Sephagen incubation using Sephadex G-50
And separated from the chemical reactants by size exclusion chromatography. Functional coagulation
The yield results based on activity are shown in Table VIII.   Example 9   Purification of human factor VIII from cryoprecipitate
Isolate a protein rich in ibronectin. Fibronectin has a pH of 6.
9 to a protein concentration of 1.34 mg / ml (33.5 mg total). Cap
Sodium acrylate was added to a concentration of 2.0% by weight, and the pH was adjusted with 1N HCl.
Readjusted to 6.9. This is the Henderson Hasselbalf at pH 6.9
Calculated from 0.014% by weight of caprylic acid and caprylate in ionized form.
986% by weight. After 1 hour incubation at 5 ° C, fibrone
Kuching was subjected to size exclusion chromatography using Sephadex G-50.
More separated from chemical reactants. Enzyme-linked immunosorbent samples before and after caprylic acid treatment
Assay by Enzyme-linked immunosorbent assay and fast
Structural changes were analyzed by protein liquid chromatography (FPLC). Implementation
The results of Example 9 are shown in Table IX.   Example 10   α₁Transferrin, also known as metal-binding globulin, is
Isolated from Fraction IV-1 from a cold ethanol purification process of Y. corn. This tran
Spherin was adjusted to a pH of 6.8 to 3.35 mg / ml (total 105 mg). Cap
Sodium acrylate was added to a concentration of 2.0% by weight and added with 1N HCl.
PH was adjusted to 6.8. This is Henderson Hasselbal at pH 6.8
Calculated that 0.022% by weight of caprylic acid and caprylate 1 in ionized form
. 978% by weight. After incubation at 5 ° C for 60 minutes,
Size exclusion chromatography of suferin using Sephadex G-50
From the chemical reactants. The sample is subjected to RID before and after caprylic acid treatment.
The yield was determined by analysis and structural changes were analyzed by FPLC. Experiment 10
The results are shown in Table X.   Example 11   Recover frozen sediment by centrifugation from a pool of freshly frozen human plasma
did. Coagulation factor VIII, also known as anti-hemophilia factor (AHF),
Recovered and purified. 1.7 units of purified AHF solution at pH 7.2
/ Ml (total 46.2 AHF units). Sodium caprylate at 5 ° C
Thorium has a concentration of 2.0% by weight And adjusted to pH 7.2 with 1N HCl.
0.0087% by weight of caprylic acid as calculated by Henderson-Hasselbarf
And 1.9913% by weight of caprylate in ionized form. 2 hours at 5 ° C
After incubation, the AHF-enriched protein was purified by Sephadex G-50.
Was separated from the chemical reactants by size exclusion chromatography. function
The yield results based on the specific factor VIII clotting activity are shown in Table XI.   Example 12   Examination of immunoglobulin M (IgM-pd) derived from human plasma purified from fraction III
Elucidated the role of low pH (pH 4.8) in virus disruption. Fraction III paste
, Cohn-Oncley cold ethanol fractionation method
ation method) from normal human plasma. Fraction III paste, pH 4
. Suspended by mixing in 0.05M sodium acetate at 0 ° C at 0 ° C.
Insoluble proteins were removed by centrifugation and filtration. IgM-pd rich clearing
The adjusted filtrate pd was adjusted to 4.8. This IgM-pd solution was brought into contact with VSV at 5 ° C.
Was. Tissue culture (TC) medium samples were contacted with VSV as a control. Virus
After piking, samples were tested for infectivity at intervals of 8 hours or less. Example 1
The results of Example 2 are shown in Table XII.   Given the foregoing disclosure and examples, those skilled in the art will be able to modify them.
Will be able to. Accordingly, the invention disclosed herein is limited only by the following claims.
It is intended to be specified.   The main features and aspects of the present invention are as follows.   1. Inactive lipid-encapsulated virus in solution of biologically active therapeutic protein
Is a method of   Virus sensitivity without adversely affecting the amount and biological activity of this protein
Contacting said solution with caprylic acid under conditions sufficient to substantially reduce dyeing power
A method comprising the steps of   2. Caprylic acid ranges from about 0.07% to about 0.001% by weight in water
The method of claim 1 wherein the amount is in non-ionized form.   3. The method of claim 1, wherein the infectivity is reduced to an undetectable level.   4. Loss, if any, of the biological activity of the processed protein
The method of claim 1 wherein the activity is less than about 30%.   5. The conditions of caprylic acid concentration and solution pH are indicated by oblique lines in the attached drawings. 2. The method according to the above 1, wherein the conditions are determined by the region.   6. The solution comprises caprylate ions and the caprylate ion concentration and
The conditions of the solution pH should be such that the concentration of caprylic acid is about 0.07% to 0.4% by weight in water.
The method of claim 1 wherein the conditions are such that they are maintained between 001%.   7. Caprylic acid concentration in the range of 0.07% to 0.01% by weight in water
2. The method according to 1 above, wherein   8. The microorganism is selected from HSV-1, VSV, vaccinia, Sindbis and EBV.
2. The method according to 1 above, wherein the virus is a lipid-coated virus.   9. Biologically active proteins include antibodies, human serum albumin, coagulation factors,
The above-mentioned 1, which is one or more of fibronectin and transferrin
The described method.   10. The clotting factor is selected from Factors II, VII, VIII, IX and X, as described in 9 above.
the method of.   11. A method for inactivating a lipid-encapsulated virus in an aqueous solution of an antibody,   For a time and pH sufficient to substantially reduce the infectivity of the virus,
% Caprylic acid and said solution in a concentration ranging from about 0.07% to about 0.001% on a% basis.
A method comprising the step of contacting   12. The antibody described in the above item 11, wherein the antibody is an IgM antibody, and the pH of the solution is about 8.0.
The method described.   13. 12. The method according to the above 11, wherein the antibody is an IgG antibody and the pH is about 6.3.
Law.   14． Lipids in aqueous solutions of biologically active human monoclonal antibodies A method for inactivating an encapsulated virus, comprising:   Time sufficient to reduce virus titer to undetectable levels, based on weight percent
At a concentration ranging from about 0.07% to about 0.001% with caprylic acid and the virus.
A method comprising contacting.   15. The antibody is a Pseudomonas aeruginosa bacterium (Pseudomonas aeruginosa bacterium)
15. The method according to the above 14, which is an antibody that binds to a serotype determinant on the lipopolysaccharide molecule of the above.   16. The bacterium is one of Fisher-Deblin immunotypes 1 to 7;
16. The method according to 15 above.   17． 17. The method according to the above 16, wherein the bacterium is Fischer immunotype 4.
.   18. The antibody is Pseudomonas aeruginosa (Pseudomonas aeruginosa)
15. The method according to the above 14, which is a body.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a graph showing the relationship between the pH of a given solution and the concentration of caprylic acid with respect to the virus inactivation time.

Claims (1)

[Claims] 1. A method for inactivating a lipid-encapsulated virus in a solution of a biologically active therapeutic protein, wherein said protein is one or more selected from antibodies, coagulation factors, fibronectin and transferrin. Contacting said solution with caprylic acid under conditions sufficient to substantially reduce the infectivity of the virus without adversely affecting the amount and biological activity of the protein. 2. A method of inactivating a lipid-encapsulated virus in an aqueous solution of an antibody, wherein the time and the pH are sufficient to substantially reduce the infectivity of the virus, from about 0.07% to Contacting said solution with caprylic acid at a concentration in the range of about 0.001%. 3. A method for inactivating a lipid-encapsulated virus in an aqueous solution of a biologically active human monoclonal antibody, wherein the virus-encapsulated virus is reduced to about 0% by weight for a time sufficient to reduce virus titer to undetectable levels. Contacting said virus with caprylic acid at a concentration ranging from 0.07% to about 0.001%.