JPS61148127A - ＨＢｃ抗原の精製方法 - Google Patents
ＨＢｃ抗原の精製方法Info
- Publication number
- JPS61148127A JPS61148127A JP59271432A JP27143284A JPS61148127A JP S61148127 A JPS61148127 A JP S61148127A JP 59271432 A JP59271432 A JP 59271432A JP 27143284 A JP27143284 A JP 27143284A JP S61148127 A JPS61148127 A JP S61148127A
- Authority
- JP
- Japan
- Prior art keywords
- antigen
- acid
- hbc
- item
- recombinant
- 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.)
- Granted
Links
- 239000000427 antigen Substances 0.000 title claims abstract description 104
- 102000036639 antigens Human genes 0.000 title claims abstract description 104
- 108091007433 antigens Proteins 0.000 title claims abstract description 104
- 238000000746 purification Methods 0.000 title claims abstract description 21
- 238000000034 method Methods 0.000 claims abstract description 57
- 239000002253 acid Substances 0.000 claims abstract description 20
- 238000004255 ion exchange chromatography Methods 0.000 claims abstract description 15
- 239000002244 precipitate Substances 0.000 claims abstract description 14
- 239000002994 raw material Substances 0.000 claims abstract description 14
- 239000000356 contaminant Substances 0.000 claims abstract description 11
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 11
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 8
- 108020004511 Recombinant DNA Proteins 0.000 claims abstract description 8
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims abstract description 7
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims abstract description 7
- 235000011130 ammonium sulphate Nutrition 0.000 claims abstract description 7
- 150000002632 lipids Chemical class 0.000 claims abstract description 6
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 6
- 238000005185 salting out Methods 0.000 claims abstract description 5
- 125000000524 functional group Chemical group 0.000 claims abstract description 4
- 238000011282 treatment Methods 0.000 claims abstract description 3
- -1 diethylaminoethyl group Chemical group 0.000 claims abstract 2
- 150000001450 anions Chemical class 0.000 claims description 18
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 12
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 claims description 9
- 238000011276 addition treatment Methods 0.000 claims description 9
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 6
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 6
- 239000000287 crude extract Substances 0.000 claims description 6
- 239000004094 surface-active agent Substances 0.000 claims description 4
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 3
- 239000011324 bead Substances 0.000 claims description 3
- 239000011521 glass Substances 0.000 claims description 3
- 235000006408 oxalic acid Nutrition 0.000 claims description 3
- 239000000463 material Substances 0.000 abstract description 11
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 5
- 239000000047 product Substances 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 4
- 230000002378 acidificating effect Effects 0.000 abstract description 3
- 208000002672 hepatitis B Diseases 0.000 abstract description 3
- 238000005349 anion exchange Methods 0.000 abstract 2
- 230000002401 inhibitory effect Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 14
- 239000000872 buffer Substances 0.000 description 12
- 238000005119 centrifugation Methods 0.000 description 11
- 239000008363 phosphate buffer Substances 0.000 description 11
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 9
- 230000001580 bacterial effect Effects 0.000 description 9
- 238000004587 chromatography analysis Methods 0.000 description 9
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 8
- 238000010828 elution Methods 0.000 description 8
- 229910052588 hydroxylapatite Inorganic materials 0.000 description 8
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 6
- 229930006000 Sucrose Natural products 0.000 description 6
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 6
- 238000000432 density-gradient centrifugation Methods 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 239000005720 sucrose Substances 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- 238000012258 culturing Methods 0.000 description 5
- 239000000499 gel Substances 0.000 description 5
- 239000013612 plasmid Substances 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 241000700605 Viruses Species 0.000 description 4
- 239000007853 buffer solution Substances 0.000 description 4
- 239000006167 equilibration buffer Substances 0.000 description 4
- 208000015181 infectious disease Diseases 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000001103 potassium chloride Substances 0.000 description 4
- 235000011164 potassium chloride Nutrition 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 108091008146 restriction endonucleases Proteins 0.000 description 4
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- 108091028043 Nucleic acid sequence Proteins 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 239000012141 concentrate Substances 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 238000000502 dialysis Methods 0.000 description 3
- 210000004185 liver Anatomy 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000008057 potassium phosphate buffer Substances 0.000 description 3
- 238000001179 sorption measurement Methods 0.000 description 3
- 241000700721 Hepatitis B virus Species 0.000 description 2
- AFCARXCZXQIEQB-UHFFFAOYSA-N N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CCNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 AFCARXCZXQIEQB-UHFFFAOYSA-N 0.000 description 2
- 229920005654 Sephadex Polymers 0.000 description 2
- 239000012507 Sephadex™ Substances 0.000 description 2
- 229920002684 Sepharose Polymers 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- AIYUHDOJVYHVIT-UHFFFAOYSA-M caesium chloride Chemical compound [Cl-].[Cs+] AIYUHDOJVYHVIT-UHFFFAOYSA-M 0.000 description 2
- 210000002421 cell wall Anatomy 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 230000002458 infectious effect Effects 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 238000005199 ultracentrifugation Methods 0.000 description 2
- 229960005486 vaccine Drugs 0.000 description 2
- 230000002747 voluntary effect Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- VZSRBBMJRBPUNF-UHFFFAOYSA-N 2-(2,3-dihydro-1H-inden-2-ylamino)-N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]pyrimidine-5-carboxamide Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C(=O)NCCC(N1CC2=C(CC1)NN=N2)=O VZSRBBMJRBPUNF-UHFFFAOYSA-N 0.000 description 1
- BFSVOASYOCHEOV-UHFFFAOYSA-N 2-diethylaminoethanol Chemical compound CCN(CC)CCO BFSVOASYOCHEOV-UHFFFAOYSA-N 0.000 description 1
- 101710145634 Antigen 1 Proteins 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- GUBGYTABKSRVRQ-WFVLMXAXSA-N DEAE-cellulose Chemical compound OC1C(O)C(O)C(CO)O[C@H]1O[C@@H]1C(CO)OC(O)C(O)C1O GUBGYTABKSRVRQ-WFVLMXAXSA-N 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 101710181478 Envelope glycoprotein GP350 Proteins 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- MZVQCMJNVPIDEA-UHFFFAOYSA-N [CH2]CN(CC)CC Chemical group [CH2]CN(CC)CC MZVQCMJNVPIDEA-UHFFFAOYSA-N 0.000 description 1
- 238000010306 acid treatment Methods 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 238000012870 ammonium sulfate precipitation Methods 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 238000007374 clinical diagnostic method Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000011067 equilibration Methods 0.000 description 1
- 239000013613 expression plasmid Substances 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012847 fine chemical Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000001738 isopycnic centrifugation Methods 0.000 description 1
- 210000003127 knee Anatomy 0.000 description 1
- 238000000464 low-speed centrifugation Methods 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 229940080469 phosphocellulose Drugs 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229940093429 polyethylene glycol 6000 Drugs 0.000 description 1
- 238000011176 pooling Methods 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 238000002333 serotherapy Methods 0.000 description 1
- 239000013605 shuttle vector Substances 0.000 description 1
- 238000000527 sonication Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical group [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 108010082737 zymolyase Proteins 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/576—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis
- G01N33/5761—Hepatitis B
- G01N33/5762—Hepatitis B core antigen
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2730/00—Reverse transcribing DNA viruses
- C12N2730/00011—Details
- C12N2730/10011—Hepadnaviridae
- C12N2730/10111—Orthohepadnavirus, e.g. hepatitis B virus
- C12N2730/10122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Urology & Nephrology (AREA)
- Biochemistry (AREA)
- Hematology (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Cell Biology (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Virology (AREA)
- Communicable Diseases (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Peptides Or Proteins (AREA)
Priority Applications (7)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP59271432A JPS61148127A (ja) | 1984-12-21 | 1984-12-21 | ＨＢｃ抗原の精製方法 |
| CA000498178A CA1268437A (en) | 1984-12-21 | 1985-12-19 | Method for purification of hbc antigen and method for measurement of hbc antibody by using said purified hbc antigen |
| KR1019850009659A KR930012113B1 (ko) | 1984-12-21 | 1985-12-20 | HBc 항원의 정재방법 및 정제된 HBc 항원을 이용한 HBc 항체의 측정방법 |
| EP85116388A EP0185391B1 (en) | 1984-12-21 | 1985-12-20 | Method for the purification of the hbc antigen and method for the measurement of hbc antibodies by using said purified hbc antigen |
| US06/811,399 US4839277A (en) | 1984-12-21 | 1985-12-20 | Method for purification of HBc antigen and method for measurement of HBc antibody by using said purified HBc antigen |
| AT85116388T ATE66679T1 (de) | 1984-12-21 | 1985-12-20 | Verfahren zur reinigung vom hbc-antigen und verfahren zum messen von hbc-antikoerpern mit diesem gereinigten hbc-antigen. |
| DE8585116388T DE3583927D1 (de) | 1984-12-21 | 1985-12-20 | Verfahren zur reinigung vom hbc-antigen und verfahren zum messen von hbc-antikoerpern mit diesem gereinigten hbc-antigen. |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP59271432A JPS61148127A (ja) | 1984-12-21 | 1984-12-21 | ＨＢｃ抗原の精製方法 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS61148127A true JPS61148127A (ja) | 1986-07-05 |
| JPH0574599B2 JPH0574599B2 (enrdf_load_stackoverflow) | 1993-10-18 |
Family
ID=17499948
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP59271432A Granted JPS61148127A (ja) | 1984-12-21 | 1984-12-21 | ＨＢｃ抗原の精製方法 |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS61148127A (enrdf_load_stackoverflow) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1990002140A1 (en) * | 1988-08-19 | 1990-03-08 | Rikagaku Kenkyusho | Method for purifying teneicine |
| JPH08504954A (ja) * | 1993-04-28 | 1996-05-28 | ラッキー リミテッド | Ｂ型およびｃ型肝炎同時診断用診断キットおよび方法 |
-
1984
- 1984-12-21 JP JP59271432A patent/JPS61148127A/ja active Granted
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1990002140A1 (en) * | 1988-08-19 | 1990-03-08 | Rikagaku Kenkyusho | Method for purifying teneicine |
| JPH08504954A (ja) * | 1993-04-28 | 1996-05-28 | ラッキー リミテッド | Ｂ型およびｃ型肝炎同時診断用診断キットおよび方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0574599B2 (enrdf_load_stackoverflow) | 1993-10-18 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| EXPY | Cancellation because of completion of term |