CN101361973A - Medicine for treating chicken pox and preparation method thereof - Google Patents
Medicine for treating chicken pox and preparation method thereof Download PDFInfo
- Publication number
- CN101361973A CN101361973A CNA2008103046444A CN200810304644A CN101361973A CN 101361973 A CN101361973 A CN 101361973A CN A2008103046444 A CNA2008103046444 A CN A2008103046444A CN 200810304644 A CN200810304644 A CN 200810304644A CN 101361973 A CN101361973 A CN 101361973A
- Authority
- CN
- China
- Prior art keywords
- solution
- chickenpox
- value
- fii
- ethanol
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 201000006082 Chickenpox Diseases 0.000 title claims abstract description 60
- 206010046980 Varicella Diseases 0.000 title claims abstract description 60
- 238000002360 preparation method Methods 0.000 title claims abstract description 23
- 239000003814 drug Substances 0.000 title claims abstract description 22
- 229940079593 drug Drugs 0.000 title abstract description 6
- 210000002381 plasma Anatomy 0.000 claims abstract description 41
- 108060003951 Immunoglobulin Proteins 0.000 claims abstract description 35
- 102000018358 immunoglobulin Human genes 0.000 claims abstract description 35
- 241000701022 Cytomegalovirus Species 0.000 claims abstract description 18
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 9
- 201000010099 disease Diseases 0.000 claims abstract description 8
- 229940021648 varicella vaccine Drugs 0.000 claims abstract description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 78
- 239000000243 solution Substances 0.000 claims description 74
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 46
- 241000700605 Viruses Species 0.000 claims description 40
- 238000001556 precipitation Methods 0.000 claims description 28
- 239000011780 sodium chloride Substances 0.000 claims description 23
- 239000000706 filtrate Substances 0.000 claims description 20
- 230000002779 inactivation Effects 0.000 claims description 19
- 239000000047 product Substances 0.000 claims description 19
- 238000001914 filtration Methods 0.000 claims description 18
- 239000000376 reactant Substances 0.000 claims description 17
- 238000000108 ultra-filtration Methods 0.000 claims description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 17
- 238000010253 intravenous injection Methods 0.000 claims description 15
- 238000003756 stirring Methods 0.000 claims description 14
- 239000008215 water for injection Substances 0.000 claims description 14
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 12
- 102000004169 proteins and genes Human genes 0.000 claims description 10
- 108090000623 proteins and genes Proteins 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 9
- 238000002844 melting Methods 0.000 claims description 8
- 230000008018 melting Effects 0.000 claims description 8
- 230000001105 regulatory effect Effects 0.000 claims description 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 7
- 239000002244 precipitate Substances 0.000 claims description 7
- 238000005349 anion exchange Methods 0.000 claims description 6
- 239000012295 chemical reaction liquid Substances 0.000 claims description 6
- 238000004587 chromatography analysis Methods 0.000 claims description 6
- 239000012141 concentrate Substances 0.000 claims description 6
- 238000007865 diluting Methods 0.000 claims description 6
- 238000005227 gel permeation chromatography Methods 0.000 claims description 6
- 238000012856 packing Methods 0.000 claims description 6
- 239000012460 protein solution Substances 0.000 claims description 6
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 claims description 5
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 5
- 238000001179 sorption measurement Methods 0.000 claims description 5
- 238000006243 chemical reaction Methods 0.000 claims description 4
- 238000005352 clarification Methods 0.000 claims description 4
- 230000036039 immunity Effects 0.000 claims description 4
- 239000012535 impurity Substances 0.000 claims description 4
- 230000002265 prevention Effects 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 4
- 238000010257 thawing Methods 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 230000001954 sterilising effect Effects 0.000 claims description 3
- 238000004659 sterilization and disinfection Methods 0.000 claims description 3
- 238000000502 dialysis Methods 0.000 claims description 2
- 239000002552 dosage form Substances 0.000 claims description 2
- 239000012528 membrane Substances 0.000 claims description 2
- 239000007974 sodium acetate buffer Substances 0.000 claims description 2
- BHZOKUMUHVTPBX-UHFFFAOYSA-M sodium acetic acid acetate Chemical compound [Na+].CC(O)=O.CC([O-])=O BHZOKUMUHVTPBX-UHFFFAOYSA-M 0.000 claims description 2
- 230000003053 immunization Effects 0.000 abstract description 5
- 239000010836 blood and blood product Substances 0.000 abstract description 2
- 229940125691 blood product Drugs 0.000 abstract description 2
- 239000004480 active ingredient Substances 0.000 abstract 1
- 239000002994 raw material Substances 0.000 abstract 1
- 238000012360 testing method Methods 0.000 description 16
- 238000000034 method Methods 0.000 description 9
- 230000009849 deactivation Effects 0.000 description 8
- 210000004369 blood Anatomy 0.000 description 7
- 239000008280 blood Substances 0.000 description 7
- 102000004190 Enzymes Human genes 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 6
- 241000701085 Human alphaherpesvirus 3 Species 0.000 description 6
- 230000001900 immune effect Effects 0.000 description 6
- 238000012937 correction Methods 0.000 description 5
- 208000015181 infectious disease Diseases 0.000 description 5
- 238000002649 immunization Methods 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 239000008363 phosphate buffer Substances 0.000 description 4
- 238000003908 quality control method Methods 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 239000000427 antigen Substances 0.000 description 3
- 102000036639 antigens Human genes 0.000 description 3
- 108091007433 antigens Proteins 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000011049 filling Methods 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 150000002632 lipids Chemical class 0.000 description 3
- 238000012372 quality testing Methods 0.000 description 3
- 239000011550 stock solution Substances 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 2
- 241000709721 Hepatovirus A Species 0.000 description 2
- 241000702617 Human parvovirus B19 Species 0.000 description 2
- 241000191967 Staphylococcus aureus Species 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000001143 conditioned effect Effects 0.000 description 2
- 238000012864 cross contamination Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 208000006454 hepatitis Diseases 0.000 description 2
- 231100000283 hepatitis Toxicity 0.000 description 2
- 230000002458 infectious effect Effects 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 229960001201 live attenuated varicella Drugs 0.000 description 2
- 244000000010 microbial pathogen Species 0.000 description 2
- 239000000178 monomer Substances 0.000 description 2
- 239000002002 slurry Substances 0.000 description 2
- 208000006379 syphilis Diseases 0.000 description 2
- 229960005486 vaccine Drugs 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 108010017384 Blood Proteins Proteins 0.000 description 1
- 206010011831 Cytomegalovirus infection Diseases 0.000 description 1
- 208000007514 Herpes zoster Diseases 0.000 description 1
- 241000701024 Human betaherpesvirus 5 Species 0.000 description 1
- 208000000474 Poliomyelitis Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000000875 corresponding effect Effects 0.000 description 1
- 229940002080 cytomegalovirus immune globulin Drugs 0.000 description 1
- 229940036098 cytomegalovirus immunoglobulin Drugs 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 239000003978 infusion fluid Substances 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000009758 senescence Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 206010040882 skin lesion Diseases 0.000 description 1
- 231100000444 skin lesion Toxicity 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention relates to a drug for treating chicken pox and a preparation method thereof, pertaining to the field of blood products. The technical problem solved by the invention is that: a drug which can effectively treat chicken pox is provided. The drug for treating chicken pox is a preparation which is prepared by adopting a chicken pox human immune globulin as a main active ingredient and adding with accessories acceptable in pharmacy, wherein, the chicken pox human immune globulin is prepared by adopting blood plasma obtained after a varicella vaccine is used for immunizing human bodies as raw material. The drug for treating chicken pox can be used for treating or preventing cytomegalovirus diseases and has wide application prospect.
Description
Technical field
The present invention relates to a kind of medicine for the treatment of chickenpox and preparation method thereof, belong to the blood products field.
Background technology
Varicella-zoster is that (varicella-zoster virus VZV) infects the disease that causes by varicella zoster virus.VZV infects disease common childhood period of being, can propagate by respiratory secretions, and infectiousness is strong.It is slight that healthy children infects many symptoms, but can cause complication such as visceral organ injury such as the infection of the unusual big conditioned pathogens such as disseminated infections, Secondary cases A group streptococcus and staphylococcus aureus of skin lesion and lung, brain, liver, kidney, the heart, eye some special population such as immunologic inadequacy person, even death.
Along with the process of organ transplantation and social population's senescence, VZV infects can be very common, also is problem demanding prompt solution.Though chickenpox vaccine can effectively prevent the generation of chickenpox, the whole world still has a lot of people not inoculate chickenpox vaccine, and therefore, the medicine that can treat chickenpox still is one of this area research emphasis.
It is chickenpox human normal immunoglobulin's intravenous injection of 25IU/ml that European Pharmacopoeia discloses specification, and adopting screening natural infection crowd convalescent period blood plasma is that feedstock production gets, and is used for the prevention of chickenpox.
At present, do not see that the relevant report that can effectively treat the medicine of chickenpox is arranged.
Summary of the invention
First technical problem to be solved by this invention provides a kind of medicine that can effectively treat chickenpox.
The medicine that the present invention treats chickenpox is to be main active with the chickenpox human normal immunoglobulin, adds the preparation that acceptable accessories is prepared from; Wherein, described chickenpox human normal immunoglobulin adopts human body to carry out that gained blood plasma is that feedstock production gets after the chickenpox vaccine immunity.
Wherein, above-mentioned human body carries out after the chickenpox vaccine immunity chickenpox virus antibody titer 〉=8IU/ml in the gained blood plasma.The source of described blood plasma is as follows:
1, use freeze-dried live attenuated varicella vaccine that the blood donor is carried out inoculation.Once more the blood donor is carried out the booster immunization injection after 3 months.
2, collect the blood plasma of blood donor's booster immunization injection after 3 months, and measure chickenpox virus antibody titer in the blood plasma, the blood plasma of chickenpox virus antibody titer 〉=8U/ml is qualified blood plasma.
Further, the present invention treats the medicine of chickenpox, and its chickenpox human normal immunoglobulin's preparation method comprises the steps:
A, blood plasma melting: with the raw blood plasma of chickenpox virus antibody titer 〉=8IU/ml in 0～4 ℃ of thawing;
B, component separate: be that 0.9% sodium chloride solution diluting plasma to protein content is 45～55g/L with the quality percentage composition, regulate pH value to 7.0 ± 0.4, adding 95% ethanol to reactant liquor concentration of alcohol is 8%, the course of reaction temperature is controlled at-2.5 ± 0.5 ℃, the centrifugalize supernatant promptly gets the FI reactant liquor;
C, filtration: the FI reactant liquor is regulated pH value to 6.80 ± 0.10, adding 95% ethanol to ethanol final concentration is 20%, and reacting liquid temperature is controlled at 5.0 ± 0.5 ℃, adds diatomite adsorption impurity, stir, leave standstill, filter, the gained precipitation is the FII+III precipitation;
D, resolution of precipitate: with of the sodium chloride solution dissolving of FII+III precipitation with 0.01mol/L, regulator solution pH value to 5.10 ± 0.05, the regulator solution ionic strength is 0.01, adding 95% ethanol concentration of alcohol to the solution is 17%, the control solution temperature is-4.5 ± 0.5 ℃, stir, leave standstill, filter, gained filtrate is FII filtrate;
E, filtrate precipitation: add sodium chloride solution in the FII filtrate, the regulator solution ionic strength is 0.05, regulator solution pH value to 6.90 ± 0.05, adding 95% ethanol concentration of alcohol to the solution is 25%, every liter of ethanol is added 1mol/L sodium chloride solution 0.05L, control end reaction liquid temp is-7.5 ± 0.5 ℃, stirs, leaves standstill, filters, and promptly gets the FII precipitation;
F, chromatography: FII precipitation is dissolved after-filtration with water for injection, and filtrate is used the anion exchange gel chromatography then with the water for injection dealcoholysis of dialysing, and obtains IgG solution;
G, ultrafiltration: with the ultrafiltration of IgG solution, 4 times of gained ultrafiltrates concentrate, and regulate with water for injection then that protein concentration is 15 ± 5g/L in the ultrafiltrate, are no more than 350uS/cm with hydrochloric acid adjusting ultrafiltrate pH value to 4.8 ± 0.1, the electrical conductivity of 0.5mol/L;
H, Pasteur's inactivation of viruses: g step gained solution was heated 10 hours in 60 ± 0.5 ℃ of water-baths continuously;
I: low pH is incubated and is put inactivation of viruses: with the protein solution behind the h step inactivation of viruses filter, clarification, ultrafiltration once more, concentrate or lyophilizing promptly.
Wherein, the dosage form of above-mentioned preparation is preferably intravenous injection.Further, the specification of described intravenous injection is 50IU/ml, 40ml.
Second technical problem to be solved by this invention provides the preparation method of above-mentioned intravenous injection, and this method comprises the steps:
A, blood plasma melting: with the raw blood plasma of chickenpox virus antibody titer 〉=8IU/ml in 0～4 ℃ of thawing;
B, component separate: be that 0.9% sodium chloride solution diluting plasma to protein content is 45～55g/L with the quality percentage composition, regulate pH value to 7.0 ± 0.4, adding 95% ethanol to reactant liquor concentration of alcohol is 8%, the course of reaction temperature is controlled at-2.5 ± 0.5 ℃, the centrifugalize supernatant promptly gets the FI reactant liquor;
C, filtration: the FI reactant liquor is regulated pH value to 6.80 ± 0.10, adding 95% ethanol to ethanol final concentration is 20%, and reacting liquid temperature is controlled at 5.0 ± 0.5 ℃, adds diatomite adsorption impurity, stir, leave standstill, filter, the gained precipitation is the FII+III precipitation;
D, resolution of precipitate: with of the sodium chloride solution dissolving of FII+III precipitation with 0.01mol/L, regulator solution pH value to 5.10 ± 0.05, the regulator solution ionic strength is 0.01, adding 95% ethanol concentration of alcohol to the solution is 17%, the control solution temperature is-4.5 ± 0.5 ℃, stir, leave standstill, filter, gained filtrate is FII filtrate;
E, filtrate precipitation: add sodium chloride solution in the FII filtrate, the regulator solution ionic strength is 0.05, regulator solution pH value to 6.90 ± 0.05, adding 95% ethanol concentration of alcohol to the solution is 25%, every liter of ethanol is added 1mol/L sodium chloride solution 0.05L, control end reaction liquid temp is-7.5 ± 0.5 ℃, stirs, leaves standstill, filters, and promptly gets the FII precipitation;
F, chromatography: FII precipitation is dissolved after-filtration with water for injection, and filtrate is used the anion exchange gel chromatography then with the water for injection dealcoholysis of dialysing, and obtains IgG solution;
G, ultrafiltration: with the ultrafiltration of IgG solution, 4 times of gained ultrafiltrates concentrate, and regulate with water for injection then that protein concentration is 15 ± 5g/L in the ultrafiltrate, are no more than 350uS/cm with hydrochloric acid adjusting ultrafiltrate pH value to 4.8 ± 0.1, the electrical conductivity of 0.5mol/L;
H, Pasteur's inactivation of viruses: g step gained solution was heated 10 hours in 60 ± 0.5 ℃ of water-baths continuously;
I: low pH is incubated and is put inactivation of viruses: with the filtration of the protein solution behind the h step inactivation of viruses, clarification, ultrafiltration once more, adjust with the hydrochloric acid of 0.5mol/L then that the protein solution pH value is 3.8～4.4, chickenpox virus antibody titer 〉=50IU/ml, adding maltose to maltose mass concentration is 10%, filtration sterilization was placed 21 days in 23～25 ℃ then;
J: finished product packing: by required specification packing, promptly.
Wherein, in a step used raw blood plasma should adopt the diagnostic reagent of state approval carry out the infectious pathogenic microorganisms mark as: hbs antigen (HBsAg), HCV antibody, HIV (1+2) antibody, syphilis, ALT etc. check inspection, and the result should be negative.Reject undesirable blood plasma, strictness prevents undesirable plasma bags and normal plasma cross-contamination in melting the slurry operating process.Should sterilize in the plasma bags surface of dress blood plasma, then broken bag, blood plasma melting.
Wherein, the b step is that 0.9% the proteic purpose of sodium chloride solution diluting plasma is the content that increases IgG in the c step precipitation with the quality percentage composition.
Wherein, regulate the used pH value regulator of pH value in b, c, the d step and can be the intravenous fluid pH value regulator of routine, be preferably acetic acid-sodium acetate buffer of pH4.0.
Wherein, preferably and not waste kieselguhr to save cost for diatomaceous adsorption effect in the c step, the kieselguhr addition is preferably every liter of solution 18g.
Wherein, the d step is fully dissolved in order to make the component I gG in the precipitation, and resolution of precipitate is selected the 0.01mol/L sodium chloride solution dissolving with 6-12 times of W/V for use.
Wherein, the e step adds the 1mol/L sodium chloride solution, and adjusts pH value to 6.90 ± 0.05, and adding 95% ethanol to final concentration is 25%, its objective is the yield that improves high-purity IgG.Regulating the used pH value regulator of pH value in the e step is the NaHCO of 1mol/L
3Solution
Wherein, the molecular cut off of the used bag filter of f step dialysis dealcoholysis is 30KD, during f step chromatography, adopts PS370 type anion exchange gel column, it with pH value 6.7 ± 0.3 phosphate buffer balance chromatographic column, the adjustment protein concentration is that 15～45g/L, pH value are 6.7 ± 0.3, last sample, and flow speed control is 40～120cm/h, collect IgG, behind the end of the sample, the remaining IgG with on the phosphate buffer washing chromatographic column merges with the IgG liquid of collecting.Gel chromatography has been removed the foreign protein and the IgA of the overwhelming majority in the solution, makes purity 〉=99%, IgG monomer and dimeric content 〉=97% of effluent IgG.
Wherein, do not add the stabilizing agent of protecting IgG during the product of h step process centre, purpose is to strengthen the inactivation of viruses effect; But Pasteur's heating means deactivation fat peplos and non-lipid-coated virus, the pH value of conventional Pasteur's heating means deactivation product is 7.0, the pH value of solution is 4.8 ± 0.1 when adopting Pasteur's heating means fire extinguishing virus among the present invention, can improve deactivation speed, its inactivation of viruses kind spectrum significantly more simple low pH is incubated the method for putting extensively (effective to non-lipid-coated virus).H step inactivation of viruses result: deactivation HIV, VSV, Sindbis and Polio-1 all 〉=4Log quantity virus.
Wherein, the used ultrafilter membrane molecular cut off of the ultrafiltration once more described in the i step is 10KD.Complete in order to ensure inactivation of virus, the i step is hanged down pH and is incubated and put inactivation of viruses, low pH incubates that to put inactivation of viruses better to HAV (hepatitis A virus, Hepatitis AVirus) and human parvovirus B19's non-lipid-coated virus inactivating efficacies such as (human parvovirus B19).
Wherein, the described specification of j step is 50IU/ml, 40ml.
The 3rd technical problem to be solved by this invention provided the purposes of said medicine in the medicine of preparation treatment or prevention cytomegalovirus disease.
Medicine of the present invention can effectively be treated chickenpox, can also be used for the treatment of or prevent cytomegalovirus disease, for the prevention and the treatment of cytomegalovirus disease provides a kind of new approach, the quiet water filling pox human normal immunoglobulin of the inventive method preparation, its quality is good, yield and the purity height (purity of IgG 〉=99%, IgG monomer and dimeric content 〉=97%), the virus safe height, have broad application prospects.
The specific embodiment
Below in conjunction with embodiment the specific embodiment of the present invention is further described, does not therefore limit the present invention among the described scope of embodiments.
Embodiment 1 chickenpox human normal immunoglobulin's preparation
1, the freeze-dried live attenuated varicella vaccine that props up of the 0.5ml/ that uses Chinese biological goods head office Changchun Biological Products Institute to produce carries out inoculation to the blood donor.Once more the blood donor is carried out the booster immunization injection after 3 months.
2, collect the blood plasma of blood donor's booster immunization injection after 3 months, and measure chickenpox virus antibody titer in the blood plasma, the blood plasma of chickenpox virus antibody titer 〉=8U/ml is qualified blood plasma.
3, produce before every bag of blood plasma adopt the diagnostic reagent of state approval to carry out infectious pathogenic microorganisms mark such as hbs antigen (HBsAg), HCV antibody, HIV (1+2) antibody, syphilis, ALT etc. to check inspection, the result should be negative.Reject undesirable blood plasma, strictness prevents undesirable plasma bags and normal plasma cross-contamination in melting the slurry operating process.
4, plasma bags surface sterilization, broken bag, blood plasma melting temperature to 0～4 ℃, carry out component by cold ethanol Protein Separation method and separate:
With 9g/L sodium chloride solution diluting plasma protein content to 45～55g/L, regulate pH value 7.0 ± 0.4, adding 95% ethanol to reactant liquor concentration of alcohol is 8%.Reacting liquid temperature is controlled at-2.5 ± 0.5 ℃.
5, the FI reactant liquor is regulated pH value 6.80 ± 0.10, adding 95% ethanol to reactant liquor concentration of alcohol is 20%.Reacting liquid temperature is controlled at 5.0 ± 0.5 ℃, adds kieselguhr, stir after-filtration, the precipitation of collection is immediately in depositing below-30 ℃.
6, after the dissolving of FI+II+III precipitate, regulate pH value 5.10 ± 0.05, adding 95% ethanol to reactant liquor concentration of alcohol is 17%, and the control reacting liquid temperature is at-4.5 ± 0.5 ℃.Stir, leave standstill after-filtration.
7, every liter of filtered solution adds 1mol/L sodium chloride solution 0.04L, conditioned reaction liquid pH value 6.90 ± 0.05, adding 95% ethanol to reactant liquor concentration of alcohol is 25%, every liter of ethanol is added 1mol/L sodium chloride solution 0.05L end reaction liquid temp and is controlled at-7.5 ± 0.5 ℃, after-filtration is left standstill in stirring, collecting precipitation.
8, the FII precipitate dissolves after-filtration with water for injection, and filtered solution is with the water for injection dealcoholysis of dialysing.Adopt the anion exchange gel chromatography,, adjust protein concentration 15～45g/L, pH6.7 ± 0.3 with pH6.7 ± 0.3 phosphate buffer balance chromatographic column, last sample, flow velocity 40～120cm/h makes the FII flow of solution pass chromatographic column, collects IgG.Behind the end of the sample,, wear the liquid merging with stream and change down operation over to the remaining IgG on the phosphate buffer washing chromatographic column.Be adsorbed on foreign protein on the chromatographic column with 1～2mol/L sodium chloride solution eluting, regenerate with 0.2～1mol/L sodium hydroxide solution.Inserts full load, regeneration use and are no more than 400 circulations.
9, ultrafiltration behind the chromatography, adjustment protein concentration 15 ± 5g/L, pH4.8 ± 0.1, electrical conductivity are imported Pasteur's deactivation jar after being no more than 350uS/cm.
10, goods were heated 10 hours in 60 ± 0.5 ℃ of water-baths continuously, with the residual Virus Pollution of deactivation possibility.
11, ultrafiltration once more after the deactivation is adjusted pH3.8～4.4, the chickenpox antibody titer is not less than 50IU/ml, and aseptic filtration in 23～25 ℃, is placed goods 21 days, hangs down pH and incubates and put inactivation of viruses.Deactivation finishes to change goods over to 2～8 ℃ of freezers and deposits.
Stock solution is purified, be chickenpox human normal immunoglobulin stock solution after the ultrafiltration, aseptic filtration.
Embodiment 2 used for intravenous injection chickenpox human normal immunoglobulin's preparation
With chickenpox human normal immunoglobulin's stock solution preparation intravenous injection of embodiment 1 preparation, make that IgG content is not less than 50g/L in the intravenous injection, the chickenpox antibody titer is not less than 50IU/ml, and add into maltose to 10%.
100 grades of clean rooms carry out packing ten thousand grades of parts, and every bottle of packing 2000IU (50IU/ml, 40ml)/bottle, contain human blood source chickenpox antibody 2000IU, tiring is not less than 50IU/ml, loading amount 40ml.
Test example 1 used for intravenous injection chickenpox human normal immunoglobulin's quality testing and result
Quiet water filling pox human normal immunoglobulin, and specification: 2000IU (50IU/ml, 40ml)/bottle, represent quantity: 2000 bottles, sampling observation quantity: 20 bottles.Test basis: with reference to quiet notes hepatitis b human immunoglobulin national registration standard YBS00102008.Quiet water filling pox human normal immunoglobulin's quality testing and the results are shown in Table 1.
Table 1 used for intravenous injection chickenpox human normal immunoglobulin's quality testing and result
Test example 2 chickenpox human normal immunoglobulin are that the specific antibody titres of target antigen is measured with the cytomegalovirus
1, test objective: because among the amount of specific antibody titres and the human normal immunoglobulin and the cell plaques behind the correlated virus count and be negative correlativing relation, therefore, if contained cytomegalovirus antibody titer can reach quiet notes cytomegalovirus human normal immunoglobulin specific antibody titres level substantially among the chickenpox human normal immunoglobulin of the present invention, can determine that chickenpox human normal immunoglobulin of the present invention can prevent and treat cytomegalovirus disease.
Measure chickenpox human normal immunoglobulin of the present invention and contain the cytomegalovirus antibody titer, with the positive contrast of quiet notes cytomegalovirus human normal immunoglobulin.
2, materials and methods
2.1 immunoglobulin:
2.1.1 chickenpox human normal immunoglobulin of the present invention (Human varicella immunoglobulin, HVIG), preproduction, lot number: 200605101, IgG content: 125.9g/L, specification: the 2ml/ bottle, detection is tired: 13.28IU/ml.
2.1.2 quiet notes cytomegalovirus human normal immunoglobulin (Human Cytomegalovirus Immunoglobulin forIntravenous Administration, HCIG), Lot:MIVCMV-198, IgG content: 52g/L, specification: 10g/ bottle expiration date: 6-19-2006, detection is tired: 76.82IU/ml.Massachusetts Public HealthBiologicLaboratories USA produces.
2.2 IgG enzyme linked immunological kit:
2.2.1 varicella zoster virus IgG enzyme linked immunological kit (Varicella Zoster Virus IgG ELISA), German IBL-Hamburg GmbH company product, lot number: VG-173.
2.2.2 cytomegalovirus IgG enzyme linked immunological kit (Cytomegalovirus IgG ELISA), Italian DIESSE company product, lot number: 142.
2.3 method:
2.3.1 test condition and operation sequence: undertaken by the description of IgG enzyme linked immunological kit respectively.
2.3.2 be subjected to the dilution factor of test product: use the diluted of IgG enzyme linked immunological kit configuration to be subjected to test product.By the primary dcreening operation test of tiring, obtain each IgG respectively and be subjected to a suitable dilution factor of test product.
2.3.3 be subjected to the OD average of test product: each identical sample joins 8 quantitative enzyme mark holes, the average of removing the highest, minimum 2 back 6 OD values of value that statistics obtains respectively.
2.3.4 standard curve logarithm regression coefficient: use the labelled amount of 5 quality-control products and OD to detect average is tried to achieve quality-control product by the logarithm recurrence detected value respectively, the labelled amount that re-uses 5 quality-control products of this equation substitution calculates corresponding correction OD value respectively, makes standard curve logarithm regression coefficient 〉=99.9%.
2.3.5 correction coefficient and corrected value: quality-control product antibody titer sign value/detected value=correction coefficient (F coefficient), differ bigger when interval if be subjected to the antibody titer detected value of test product to be positioned at two F coefficients, adopt the adjacent sign valence value sum of two adjacent detected value correction coefficient sums of tiring/two * this be subjected to test product the detection valence value=this is subjected to the F coefficient of test product.Each is subjected to the detected value * correction coefficient of the tiring=corrected value (F value) of test product.
3, result of the test
Result of the test sees Table 2.
The antigenic specific antibody titres measurement result of table 2 chickenpox, cytomegalovirus immune globulin and respective target
As can be seen from Table 2, the contained cytomegalovirus antibody titer of chickenpox human normal immunoglobulin of the present invention and cytomegalovirus human normal immunoglobulin close (be respectively 877,1477mIU/mg); In addition, the contained chickenpox virus antibody titer of chickenpox human normal immunoglobulin and cytomegalovirus human normal immunoglobulin is (be respectively 105.5,100mIU/mg) quite, so determine that chickenpox vaccine can bring out human body and produce cytomegalovirus antibody.Infer that in view of the above chickenpox human normal immunoglobulin of the present invention can prevent and treat the cytomegalovirus infection disease.
Claims (10)
- [claim 1] a kind of medicine for the treatment of chickenpox is characterized in that: it is to be main active with the chickenpox human normal immunoglobulin, adds the preparation that acceptable accessories is prepared from; Wherein, described chickenpox human normal immunoglobulin adopts human body to carry out that gained blood plasma is that feedstock production gets after the chickenpox vaccine immunity.
- The medicine of [claim 2] treatment chickenpox according to claim 1 is characterized in that: described human body carries out after the chickenpox vaccine immunity chickenpox virus antibody titer 〉=8IU/ml in the gained blood plasma.
- The medicine of [claim 3] treatment chickenpox according to claim 2 is characterized in that: described chickenpox human normal immunoglobulin's preparation method comprises the steps:A, blood plasma melting: with the raw blood plasma of chickenpox virus antibody titer 〉=8IU/ml in 0～4 ℃ of thawing;B, component separate: be that 0.9% sodium chloride solution diluting plasma to protein content is 45～55g/L with the quality percentage composition, regulate pH value to 7.0 ± 0.4, adding 95% ethanol to reactant liquor concentration of alcohol is 8%, the course of reaction temperature is controlled at-2.5 ± 0.5 ℃, the centrifugalize supernatant promptly gets the FI reactant liquor;C, filtration: the FI reactant liquor is regulated pH value to 6.80 ± 0.10, adding 95% ethanol to ethanol final concentration is 20%, and reacting liquid temperature is controlled at 5.0 ± 0.5 ℃, adds diatomite adsorption impurity, stir, leave standstill, filter, the gained precipitation is the FII+III precipitation;D, resolution of precipitate: with of the sodium chloride solution dissolving of FII+III precipitation with 0.01mol/L, regulator solution pH value to 5.10 ± 0.05, the regulator solution ionic strength is 0.01, adding 95% ethanol concentration of alcohol to the solution is 17%, the control solution temperature is-4.5 ± 0.5 ℃, stir, leave standstill, filter, gained filtrate is FII filtrate;E, filtrate precipitation: add sodium chloride solution in the FII filtrate, the regulator solution ionic strength is 0.05, regulator solution pH value to 6.90 ± 0.05, adding 95% ethanol concentration of alcohol to the solution is 25%, every liter of ethanol is added 1mol/L sodium chloride solution 0.05L, control end reaction liquid temp is-7.5 ± 0.5 ℃, stirs, leaves standstill, filters, and promptly gets the FII precipitation;F, chromatography: FII precipitation is dissolved after-filtration with water for injection, and filtrate is used the anion exchange gel chromatography then with the water for injection dealcoholysis of dialysing, and obtains IgG solution;G, ultrafiltration: with the ultrafiltration of IgG solution, 4 times of gained ultrafiltrates concentrate, and regulate with water for injection then that protein concentration is 15 ± 5g/L in the ultrafiltrate, are no more than 350uS/cm with hydrochloric acid adjusting ultrafiltrate pH value to 4.8 ± 0.1, the electrical conductivity of 0.5mol/L;H, Pasteur's inactivation of viruses: g step gained solution was heated 10 hours in 60 ± 0.5 ℃ of water-baths continuously;I: low pH is incubated and is put inactivation of viruses: with the protein solution behind the h step inactivation of viruses filter, clarification, ultrafiltration once more, concentrate or lyophilizing promptly.
- The medicine of [claim 4] treatment chickenpox according to claim 1 is characterized in that: the dosage form of described preparation is an intravenous injection.
- The medicine of [claim 5] treatment chickenpox according to claim 4 is characterized in that: the specification of described intravenous injection is 50IU/ml, 40ml.
- The preparation method of the described intravenous injection of [claim 6] claim 4 is characterized in that: comprise the steps:A, blood plasma melting: with the raw blood plasma of chickenpox virus antibody titer 〉=8IU/ml in 0～4 ℃ of thawing;B, component separate: be that 0.9% sodium chloride solution diluting plasma to protein content is 45～55g/L with the quality percentage composition, regulate pH value to 7.0 ± 0.4, adding 95% ethanol to reactant liquor concentration of alcohol is 8%, the course of reaction temperature is controlled at-2.5 ± 0.5 ℃, the centrifugalize supernatant promptly gets the FI reactant liquor;C, filtration: the FI reactant liquor is regulated pH value to 6.80 ± 0.10, adding 95% ethanol to ethanol final concentration is 20%, and reacting liquid temperature is controlled at 5.0 ± 0.5 ℃, adds diatomite adsorption impurity, stir, leave standstill, filter, the gained precipitation is the FII+III precipitation;D, resolution of precipitate: with of the sodium chloride solution dissolving of FII+III precipitation with 0.01mol/L, regulator solution pH value to 5.10 ± 0.05, the regulator solution ionic strength is 0.01, adding 95% ethanol concentration of alcohol to the solution is 17%, the control solution temperature is-4.5 ± 0.5 ℃, stir, leave standstill, filter, gained filtrate is FII filtrate;E, filtrate precipitation: add sodium chloride solution in the FII filtrate, the regulator solution ionic strength is 0.05, regulator solution pH value to 6.90 ± 0.05, adding 95% ethanol concentration of alcohol to the solution is 25%, every liter of ethanol is added 1mol/L sodium chloride solution 0.05L, control end reaction liquid temp is-7.5 ± 0.5 ℃, stirs, leaves standstill, filters, and promptly gets the FII precipitation;F, chromatography: FII precipitation is dissolved after-filtration with water for injection, and filtrate is used the anion exchange gel chromatography then with the water for injection dealcoholysis of dialysing, and obtains IgG solution;G, ultrafiltration: with the ultrafiltration of IgG solution, 4 times of gained ultrafiltrates concentrate, and regulate with water for injection then that protein concentration is 15 ± 5g/L in the ultrafiltrate, are no more than 350uS/cm with hydrochloric acid adjusting ultrafiltrate pH value to 4.8 ± 0.1, the electrical conductivity of 0.5mol/L;H, Pasteur's inactivation of viruses: g step gained solution was heated 10 hours in 60 ± 0.5 ℃ of water-baths continuously;I: low pH is incubated and is put inactivation of viruses: with the filtration of the protein solution behind the h step inactivation of viruses, clarification, ultrafiltration once more, adjust with the hydrochloric acid of 0.5mol/L then that the protein solution pH value is 3.8～4.4, chickenpox virus antibody titer 〉=50IU/ml, adding maltose to maltose mass concentration is 10%, filtration sterilization was placed 21 days in 23～25 ℃ then;J: finished product packing: by required specification packing, promptly.
- The preparation method of [claim 7] intravenous injection according to claim 6 is characterized in that: regulating the used pH value regulator of pH value in b, c, the d step is the acetic acid sodium acetate buffer of pH4.0; Regulating the used pH value regulator of pH value in the e step is the NaHCO of 1mol/L 3Solution.
- The preparation method of [claim 8] intravenous injection according to claim 6 is characterized in that: diatomaceous addition is every liter of solution 18g in the c step.
- The preparation method of [claim 9] intravenous injection according to claim 6 is characterized in that: the molecular cut off of the used bag filter of f step dialysis dealcoholysis is 30KD; The used ultrafilter membrane molecular cut off of ultrafiltration once more described in the i step is 10KD.
- The purposes of each described medicine of [claim 10] claim 1～5 in the medicine of preparation treatment or prevention cytomegalovirus disease.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008103046444A CN101361973A (en) | 2008-09-24 | 2008-09-24 | Medicine for treating chicken pox and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008103046444A CN101361973A (en) | 2008-09-24 | 2008-09-24 | Medicine for treating chicken pox and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101361973A true CN101361973A (en) | 2009-02-11 |
Family
ID=40388698
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2008103046444A Pending CN101361973A (en) | 2008-09-24 | 2008-09-24 | Medicine for treating chicken pox and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101361973A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103554252A (en) * | 2013-11-15 | 2014-02-05 | 同路生物制药股份有限公司 | Giant cell human immunoglobulin and preparation method thereof |
| CN105601735A (en) * | 2016-01-28 | 2016-05-25 | 哈尔滨派斯菲科生物制药股份有限公司 | Intravenously injected cytomegalovirus human immune globulin and preparation method thereof |
| CN105601736A (en) * | 2016-01-28 | 2016-05-25 | 哈尔滨派斯菲科生物制药股份有限公司 | Respiratory syncytial virus resistance human immune globulin and preparation method thereof |
| CN105669860A (en) * | 2016-01-28 | 2016-06-15 | 哈尔滨派斯菲科生物制药股份有限公司 | Human immunoglobulin resisting hand-foot-and-mouth disease and preparation method of human immunoglobulin |
-
2008
- 2008-09-24 CN CNA2008103046444A patent/CN101361973A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103554252A (en) * | 2013-11-15 | 2014-02-05 | 同路生物制药股份有限公司 | Giant cell human immunoglobulin and preparation method thereof |
| CN103554252B (en) * | 2013-11-15 | 2016-03-30 | 同路生物制药股份有限公司 | A kind of Giant cell human immunoglobulin and preparation method thereof |
| CN105601735A (en) * | 2016-01-28 | 2016-05-25 | 哈尔滨派斯菲科生物制药股份有限公司 | Intravenously injected cytomegalovirus human immune globulin and preparation method thereof |
| CN105601736A (en) * | 2016-01-28 | 2016-05-25 | 哈尔滨派斯菲科生物制药股份有限公司 | Respiratory syncytial virus resistance human immune globulin and preparation method thereof |
| CN105669860A (en) * | 2016-01-28 | 2016-06-15 | 哈尔滨派斯菲科生物制药股份有限公司 | Human immunoglobulin resisting hand-foot-and-mouth disease and preparation method of human immunoglobulin |
| CN105601735B (en) * | 2016-01-28 | 2019-04-19 | 哈尔滨派斯菲科生物制药股份有限公司 | A kind of intravenous Giant cell human immunoglobulin and preparation method thereof |
| CN105601736B (en) * | 2016-01-28 | 2019-04-23 | 哈尔滨派斯菲科生物制药股份有限公司 | A kind of anti respiratory syncytial virus human immunoglobulin(HIg) and preparation method thereof |
| CN105669860B (en) * | 2016-01-28 | 2019-11-22 | 哈尔滨派斯菲科生物制药股份有限公司 | A kind of hand-foot-and-mouth disease resistant human immunoglobulin and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101402944B (en) | EV-71 virus seed, inactivated vaccine for human and method of producing the same | |
| CN105601736B (en) | A kind of anti respiratory syncytial virus human immunoglobulin(HIg) and preparation method thereof | |
| CN111471103A (en) | Heterologous antibody of new coronavirus (2019-nCOV) and preparation method thereof | |
| CN105601735B (en) | A kind of intravenous Giant cell human immunoglobulin and preparation method thereof | |
| CN101361973A (en) | Medicine for treating chicken pox and preparation method thereof | |
| CN104231075B (en) | Preparation process of human hepatitis B immunoglobulin | |
| US20130172536A1 (en) | Intravenous Cytomegalovirus Human Immune Globulin and Manufacturing Method Thereof | |
| CN104001172B (en) | A kind of preparation technology of hepatitis b human immunoglobulin for intravenous injection | |
| CN115010804A (en) | A kind of production method and equipment for online separation of high-purity immunoglobulin | |
| US20220267419A1 (en) | Human Immunoglobulin Against Methicillin-Resistant Staphylococcus Aureus, Preparation Method Therefor, And Use Thereof | |
| CN102190725A (en) | Hand-foot-and-mouth disease resistant human immunoglobulin, and preparation and using methods and application thereof | |
| MX2009002909A (en) | Ipv-dpt vaccine. | |
| CN101591393B (en) | Hydrophobia-tetanus double titer human immunoglobulin, method for preparing same and application thereof in pharmacy | |
| CN112010968B (en) | Method for rapidly extracting blood plasma of patient with COVID-19 in convalescence stage for preparing immunoglobulin G | |
| CN101306200B (en) | Preparation method of hepatitis b human immunoglobulin for intravenous injection | |
| CN112225799B (en) | Method for rapidly extracting blood plasma of COVID-19 patient in recovery period by automatic separation system | |
| CN107174658B (en) | Varicella virus inactivated vaccine for human and preparation method thereof | |
| US10358462B2 (en) | Method for the preparation of immunoglobulins | |
| CN112375142A (en) | Preparation method of novel coronavirus human immunoglobulin for intravenous injection | |
| CN101927011A (en) | Virus inactivation method for solution of globulin | |
| CN113278067B (en) | Preparation method of novel coronavirus porcine immunoglobulin | |
| CN110339351A (en) | The stabilizer that a kind of preparation method of veterinary rabies inactivated vaccine and the vaccine include | |
| CN101497649B (en) | Production process of high fusion rate non-bacterial virus newborn bovine serum | |
| CN101352571A (en) | A kind of medicine for treating Japanese encephalitis and preparation method thereof | |
| CN109432413A (en) | A kind of russian spring-summer encephalitis virus inactivated vaccine and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Open date: 20090211 |