AU636145B2 - New human parvovirus peptides with disulfide bridge for immunization or diagnosis - Google Patents
New human parvovirus peptides with disulfide bridge for immunization or diagnosis Download PDFInfo
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- AU636145B2 AU636145B2 AU55596/90A AU5559690A AU636145B2 AU 636145 B2 AU636145 B2 AU 636145B2 AU 55596/90 A AU55596/90 A AU 55596/90A AU 5559690 A AU5559690 A AU 5559690A AU 636145 B2 AU636145 B2 AU 636145B2
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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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
-
- 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
- C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
- C12N2750/00011—Details
- C12N2750/14011—Parvoviridae
- C12N2750/14211—Erythrovirus, e.g. B19 virus
- C12N2750/14222—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Virology (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Description
OPI DATE 29/11/90
P
AOJP DATE 10/01/91
INTERNATI'
APPLN. ID 55596 REATY (PCT) PCT NUMBER PCT/SE90/00276 (51) International Patent Classification 5 C07K 7/10, G01N 33/569 A61K 39/23 Al (11) International Publication Number: WO 90/13567 (43) International Publication Date: 15 November 1990(15.11.90) (21) International Application Number: (22) International Filing Date: Priority data: 8901566-3 28 April PCT/SE90/00276 25 April 1990(25.04.90) 1989 (28.04.89) 71) Applicant (for all designated States except US): FERRING DIAGNOSTICA AB [SE/SE]; Soldattorpsvagen 5, S- 216 13 Malm6 (SE).
(72) Inventors; and Inventors/Applicants (for US only) TROJNAR, Jerzy [SE/ SE]; Stendregatan 36, S-230 44 Vintrie WAHREN, Britta [SE/SE]; Fritiofsvagen 10, S-182 64 Djursholm FRIDELL, Eva [SE/SE]; Slanbarsstigen 10, S-125 31 Alvsj6 (SE).
(74) Agent: AWAPATENT AB; Box 5117, S-200 71 Malmb
(SE).
(81) Designated States: AT (European patent), AU, BE (European patent), CA, CH (European patent), DE (European patent), DK (European patent), ES (European patent), FI, FR (European patent), GB (European patent), IT (European patent), JP, KR, LU (European patent).
NL (European patent), NO, SE (European patent), US.
Published With international search report.
636145 (54)Title: NEW HUMAN PARVOVIRUS PEPTIDES WITH DISULFIDE BRIDGE FOR IMMUNIZATION OR DIAG-
NOSIS
(57) Abstract An artificial peptide having an amino acid sequence which corresponds to a naturally occurring amino acid sequence of a human parvovirus comprising an epitope and which further has two cysteine residues located on each side of said epitope, and further having a sulphur bridge between said two cysteine residues, which has been formed by a chemical oxidation step, is described. Furthermore, an artificial antigen which reacts with antibodies induced by a human parvovirus is described. Said antigen mainly consists of an artificial peptide according to the invention. Additionally, a method of detecting antibodies induced by a human parvovirus in a sample of body fluid, wherein said sample is subjected to an immunoassay, especially ELISA, and wherein an artificial antigen according to the invention is used as a diagnostic antigen, is described. A diagnostic immunoassay kit for said method is also described. Finally, a vaccine composition comprising, as an immunizing component, at least one antigen according to the invention, is described.
See back of page WO 0/13567 PCY/SE90/00276 .v 1 NEW HUMAN PARVOVIRUS PEPTIDES WITH DISULFIDE BRIDGE FOR IMMUNIZATION OR DIAGNOSIS The present invention relates to artificial peptides having an amino acid sequence which corresponds to a naturally occurring amino acid sequence of a human parvovirus comprising an epitope and which further has two cysteine residues located on each side of said epitope, and further having a sulphur bridge between said two cysteine residues which has been formed by a chemical oxidation step. The invention also relates to artificial antigens, which react with antibodies induced by a human parvovirus, a method of detecting antibodies induced by a human parvovirus in a sample of body fluid, a diagnostic immunoassay kit for said method, and a vaccine composition comprising, as an immunizing component, at least one antigen of the invention.
Background A human parvovirus, B19, gives rise to erythema infectiosum in children. It is also associated with aplastic crisis in patients with chronic hemolytic sickle cell) anemia and with spontaneous abortion intrauterine death or hydrops fetalis when the woman aquires the infection during pregnancy. The fetus infection might be cured if the diagnosis is made early. (J Virol 58: 921 936, 1986; Shade et al).
The virus can only be cultivated in human bonemarrow, which hampers isolation for serology. Peptide-based serology would therefore be a means of diagnosis.
One common way to establish a diagnosis through antibody detection is to screen serum samples by enzyme- -linked immunosorbent assay (ELISA) (Sarngadharan, M.G., Popovic, Bruch, Schtpbach, J. and Gallo, R.C.) Wells of microplates coated with viral antigens are reacted with the serum samoles under investigation, washed, and antihuman Ig added. The latter reagent WO 90/13567 PCT/SE90/00276 2 is labelled with an enzyme. After washing, the enzyme labelled antihuman Ig remains only if specific antiviral Ig wa' present in the serum sample. It is visualized by addition of a substrate for the enzyme and the color reaction quantified in a spectrophotometer.
No assay has so far been developed for detection of antibodies induced by human parvovirus in a sample of body fluid.
Description of the invention The present invention provides i.a. a rapid, sensitive and specific assay for detection of antibodies induced by a human parvovirus.
In one aspect of the invention there is provided an artificial peptide having an amino acid sequence which corresponds to a naturally occurring amino acid sequence of a human parvovirus comprising an epitope and which further has two cysteine residues located on each side of said epitope, and further having a sulphur bridge between said two cysteine residues which has been formed by a chemical oxidation step.
It is believed that this stabilization of the peptide by a sulphur bridge between two cysteine residues is responsible for the useful properties of the peptide, such as an enhancement of the antibody binding activity, as well as the chemical stability of the final product.
The artificial peptide includes at least two cysteine residues, which are cyclized to form a sulphur bridge. The two cysteine residues which are linked together may have one or more amino acid residues comprising an epitope between themselves, such as 2 to 10 residues. If the artificial peptide according to the invention includes more than two cysteine residues, still only one sulphur bridge between two cysteine residues is formed by a chemical oxidation step.
In a preferred embodiment of this aspect of the invention there is provided an artificial peptide having an amino acid sequence which corresponds to a naturally occurring amino acid sequence of a human parvovirus comprising an epitope and which further has two cysteine residues located on each side of said epitope, characterised in that it 2a WO 90/13567 PCT/SE90/0076 is chosen from the group consisting of the peptide having the modified amino acid sequence H-Phe-Ser-Pro-Ala-Ala-Ser-Ser-Cys-His-Asn-Ala-Ser- -Gly-Lys-Glu-Ala-Lys-Val-Cys-Thr-Ile-Ser-Pro-Ile-NH 2 and peptides having a shorter sequence thereof including the modified sequence -Cys-His-Asn-Ala-Ser-Gly-Lys-Glu-Ala-Lys-Val-Cys-.
and the peptide having the modified amino acid sequence H-Gly-Lys-Glu-Ala-Lys-Val-Cys-Thr-Ile-Ser-Pro-Ile- -Cys-Gly-Tyr-Ser-Thr-Pro-Trp-Arg-Tyr-Leu-NH 2 -j and peptides having a shorter sequence thereof including the modified sequence -Cys-Thr-Ile-Ser-Pro-Ile-Cys-; I j In another aspect of the invention there is provided an artificial antigen which reacts with antibodies induced by a human parvovirus, which antigen mainly consists of an artificial peptide having an amino acid sequence which corresponds to a naturally occurring amino acid sequence of a human parvovirus comprising an epitope and which further has two cysteine residues located on each side of said epitope, and further having a sulphur bridge between said two cysteine
I
WO 90/135 67 PCJT/SE90/00276 4 residues which has been formed by a chemical oxidation step.
In the specification and claims the expression "antigen mainly consists of an artificial peptide" indicates that the ability of the antigen to react with antibodies derives from the artificial peptide.
In a preferred embodiment of this aspect of the invention there is provided an artificial antigen which reacts with antibodies induced by a human parvovirus, which antigen mainly consists of a preferred artificial peptide according to the invention, exemplified above.
The artificial antigens according to the invention can be immobilized or coupled to a carrier, such as mineral carriers, e.g. aluminium hydroxide, calcium phosphate, etc., plastic surfaces, e.g. microplates, beads, etc., proteins, such as bovine serum albumin or an immunizing component, such as keyhole limpet haemocyanin.
Even though the artificial antigens according to the invention so far only have been used as diagnostic antigens to detect antibodies induced by a human parvovirus, in a sample of body fluid, it is believed that they can be used as immunizing components in vaccine compositions against a human parvovirus.
Thus, a further aspect of the invention provides a vaccine composition, which comprises as an immunizing component, at least one antigen selected from artificial antigens according to the invention, together with a nontoxic pharmaceutically acceptable carrier and/or diluent.
In yet another aspect of the invention there is provided a method of detecting antibodies induced by a human parvovirus in a sample of body fluid, wherein said sample is subjected to an immunoassay and an artificial antigen according to the invention is used as a diagnostic antigen. Examples of useful body fluids are urine, saliva, tear fluid, milk, serum, blood and cerebrospinal fluid.
WO 90/13567 PP/SE90/00276 The immunoassay in which the artificial antigens according to the invention can be used as diagnostic antigens is any immunoassay of choice, such as enzymelinked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunodiffusion or immunoelectrophoreses (IE).
In a preferred embodiment of this aspect of the invention ELISA is used as the immunoassay of choice.
In a further aspect of the invention the-re is :rovided a diagnostic immunoassay kit for the detection of antibodies induced by a human parvoviru in a sample of body fluid, wherein an artificial antigen according to the invention is included as a diagnostic antigen.
Depending on the immunoassay to be used, the kit will comprise other items, such as a positive standard serum sample, a negative standard serum sample, in the case of ELISA an enzyme conjugate and optionally a substrate for the enzyme conjugate, and also optionally buffer solution(s) and/or washing solution(s). Optionally all the reagents in the kit are contained in separate sealed test tubes or vials marked with specific labels.
Synthesis of the artificial peptides of the invention The artificial peptides of the invention can be produced by any known method of producing a linear peptide sequence, such as cloning, degradation, coupling of one amino acid residue to the next one in liquid phase or coupling the amino acids to one another starting with a solid phase (resin) to which the C-terminal of the first amino acid is coupled, whereupon the C-terminal of the next amino acid is coupled to the N-terminal of the first amino acid, etc., finally releasing the built-up peptide from the solid phase (so-called Merrifield synthesis). Once the appropriate linear peptide sequence is ready, it is subjected to a chemical oxidation step in order to cyclize two cysteine residues thereof, whereby a sulphur bridge is formed between the cysteine residues.
General description of synthesis In the examples below, all peptides were synthesized on an Applied Biosystems 430A Peptide Synthesizer WO 90/13567 PCTSE90/0076 6 using a double coupling program with termination step after the second coupling. The resin used was of 4-methylbenzhydrylamine type with theoretical loading of 0.66 meq/g (Peptides International, Louisville, KY, USA).
The final product of the synthesis was dried in vacuo over night. After drying the peptide-resin was suspended in methanol (70 ml) and saturated with ammonia. The mixture was placed in pressurized steel vessel and left over night with magnetic stirring. The resin was then separated by filtration, washed several times with methanol and thoroughly dried in vacuo. The peptide was then cleaved from the resin by treatment with liquid hydrogen fluorid in the presence of anisole and ethyl-methyl-sulfide as scavangers (HF:anisole:EMS 10:05:05). After removal of hydrogen fluoride by evaporation the residue was suspended in ethyl acetate (100 ml) and filtered. The solid was washed on filter with additional ethyl acetate (3x100 ml) and the cleaved peptide extracted with acetic acid (100 ml). The extract was promptly diluted to the volume of 2 dm 3 with acetic acid in methanol and treated with 0.1 M solution of iodine in methanol until the faint brown colour persisted. Then the Dowex 1x8 ion exchanger in acetate form was added (3 g) (Bio-Rad, Richmond, CA and the mixture was filtered. The filtrate was evaporated and the residue freeze-dried from 1% acetic acid in water. The product was then purified by reversed phase liquid chromatography on a column filled with Vydac 20-25 p (Separation Group, CA) in a suitable system containing acetonitrile in 0.1% trifluoroacetic acid water solution. The samples collected from the column were analyzed by analytical HPLC Varian 5500 (Sunnyvale, CA) equipped with Bondapak C18 column (Millipore, Milford, Mass.). Fractions containing pure substance were pooled, the solvent was evaporated and the product freeze-dried from 1% acetic acid in water.
The final HPLC analysis was performed on ready product and the structure of the peptide was confirmed by amino acid analysis and FAD-MS (Fast atom bombardment spectrometry).
All amino acids used during the synthesis were protected with tert-butoxycarbonyl group at ax-amino-function.
The side chain protections used were as follows: Ser(BZL), Thr(BZL), Tyr(2-BrCbz), Lys(2-ClCbz), Glu(BZL), Arg(Tos), Cys(Mob), wherein BZL =ben/,yl io2-BrCbz =2-bromocarbobenzoxy 2-ClCbz =2-chlorocarbobenzoxy Tos =tosyl Mob =4-methoxybenzyl 6a WO 90/13567 PCT/SE90/00276 7 The final HPLC analysis was performed on ready produ< and the structure of the peptide was confirmed y amino acid analysis and FAB-MS (Fast atom bombardment spectrometry).
All amino acids used during the synthesis were protected with tert-butoxycarbonyl group at a-aminofunction. The side chain protections used were as follows: Ser(BZL), Thr(BZL), Tyr(2-BrCbz), Lys(2-ClCbz), Qlu(BZL), Arg(Tos), Cys(Mob).
Amino acid derivatives were delivered by Bachem AG, Switzerland.
The genome of human parvovirus B19 encodes two structural proteins. h.ie sequences corresponding to VP2 were first synthesized as linear peptides and then a sulphur bridge between two cysteine residues were formed by a chemical oxidation step.
EXAMPLE I (The sequence corresponds to a sequence found in B19's (parvovirus) VP2 region).
H-Phe-Ser-Pro-Ala-Ala-Ser-Ser-Cys-His-Asn-Ala-Ser-Gly- -Lys-Glu-Ala-Lys-Val-Cys-Thr-Ile-Ser-Pro-Ile-NH 2 The peptide was prepared according to the general description of synthesis. The structure was confirmed by aminoacid analysis and by FAB-MS.
EXAMPLE II (The sequence corresponds to a sequence found in B19's (parvovirus) VP2-region).
H-Gly-Lys-Glu-Ala-Lys-Val-Cys-Thr-Ile-Ser-Pro-Ile-Cys- \j -Gly-Tyr-Ser-Thr-Pro-Trp-Arg-Tyr-Leu-Ni 2 WO 90/13567 PC/SE90/00276 8 The peptide was prepared according to the general description of synthesis. The structure was confirmed by aminoacid analysis and by FAB-MS.
Detection of antibodies induced by human parvovirus in blood samples For the detection of antibodies induced by human parvovirus B19 in blood samples use was made of ELISA.
(Engvall, E. and Perlmann, Enzyme-linked immunosorbent assay (ELISA). III Quantification of specific antiboides by enzyme labelled anti-immunoglobulin in antigen-coated tubes. J. Immunol. 109.129-135,1972).
Materials: Plates: Nunc 96 F, Roskilde, Denmark Conjugate: Alkaline phosphatase labelled anti-human Ig, Sigma Buffers: Carbonate buffer: 0.05 M sodium carbonate biffer, pH approx. Buffer A: Trypsinization buffer without Ca and Mg and with bovine serum albumin (BSA) and 0.05% Tween Diethanolamine, Sigma.
Washing soltion: 0.9% NaC1 with 0.05% Tween Methods: Coating: A solution of the coating antigen (test peptide), 5 pg to 20 pg per ml, is made in carbonate buffer. 100 pl of the solution is added to each well of stripes or of 96-well microplates. The adsorption takes place over night, 18 hrs, at room temperature.
The coated plates can be stored with their contents in 4°C until use.
Serum assay: 1. Empty and wash the plate 4 times with washing solution.
2. Add 100 pl of serum diluted 1:100 in buffer A.
3. Add 100 pl of just buffer A to one well; this well serves as a blank.
WO 90/13567 PC'F/SE90/00276 9 4. Incubate 2 h at 37 0 C. (Dilute the conjugate during this period, see 6).
Wash 4 times with washing solution.
Empty.
6. Add 100 pl of a conjugate, e.g. Alkaline phosphatase labelled anti-human IgG, Sigma diluted 1:1500 in buffer A.
7. Incubate at 37 0 C for 2 h. (Prepare substrate solution during this period, see 9).
8. Wash 4 times with washing solution.
Empty.
9. Adi. 100 pl Paranitrophenylphosphate (2 tablets/10 ml diethanolamine).
Use a clean vessel. 5 min are needed to dissolve the tablets.
Incubate 30 min at room temperature.
Stop the reaction with 25 pl 5 M NaOH.
11. Read plate at 405 nm.
Using the above described materials and methods the following materials were tested as diagnostic antigens (coating antigens) for the detection of antibodies induced by human parvovirus B19 in serum samples from infected patients: A. A region of linear peptides corresponding to a sequence found in the VP2-region of human parvovirus B19. They gave absorbance values of 0.2-0.4 with known seropositive samples.
B. An artificial peptide according to the invention described in Example I of this specification.
This peptide gave absorbance values of 1.55 0.45 when the test was performed on serum samples from seropositive persons and absorbance values of WO 90/13567 PCT/SE90/00276 0,30 0.15 when the test was performed on serum samples from 9 seronegative persons.
Thus, when the antigen in the test was the peptide according to the invention the absorbance values of seropositive sera became significant, and the peptide showed good reactivity with antibodies induced by human parvovirus B19.
Claims (9)
1. An- artificial peptide having an amino acid se- quence 'which corresponds to a naturally occurring amino acid sequence of a human parvovirus com'prising an epitope and which further has two cysteine residues located on each side of said epitope, c h a r a c t, e r i s e d in that it is chosen from the group consisting of the peptide having the modified amino acid sequence H-Phe-Set -Pro-Al a-Ala--Ser-Ser-Cys-Hi s-Asn-Al a-Ser-Gly- I -Lys-Glu-Ala-Lys-Val -Cys-Thr-Ile-Ser-Pro-Ile-NH 2 and peptides having a shorter sequence thereof including the modified sequence -Cys-His-Asn-Ala-Ser-Gly-Lys-Glu-Ala-Lys-Val -Cys-; I I and the peptide having the modified amino acid sequence H-Gly-Lys-Glu-Ala-Lys-Val-Cys-Thr-Ile-Ser-Pro-Ile-Cys- Gly -Tyr -Se r -Th r -Pro -T r p-Arg -Ty r -Leu N H 2 and peptides having a shorter sequence thereof includi-g the modified sequence -Cys-Thr-Ile-Ser-Pro-Ile-Cys-. L I S' 3ST I TU T E 12PC71"7 1991 -08- 1
2. An artificial antigen which reacts with anti- bodies induced by a human parvovirus, ch a r act er i s ed in that the antigen mainly consists ofl an artificial peptide chosen from the group consisting of the peptide having the modified amino acid sequence H-Phe-Ser-Pro-Ala-Al a-Ser-Ser-Cys-His-Asn-AIla-Ser-Gl y- -Lys-Glu-Ala-Lys-Val-Cys-Thr-I le-Ser-Pro-Ile-NH 2 and peptides having a shorter sequence thereof including the modified sequence -Cys -His -As n-Al a-Se r-Gl y-Lys-G u-Al a-Lys-Val1-Cys-; and the peptide having the modified amino acid sequence H-G y-Lys-G u-Al a-Lys -Val -Cys-Thr-I I -Se,.-Pro- 11e-Cys -Gl y-Tyr-Ser-Thr-Pro-Trp-Arg-TyrLeuNH 2 and peptides having a shorter sequence thereof including the modified sequence -Cys-Thr-Ile-Ser-Pro-I le-Cys-. I I x 13 "Ik S I" PC 1991 -08- 1
3. An artificial antigen according to claim 2, c h a r a c t e r i z e d in that it has been immobilized or coupled to a carrier.
4. Method of detecting antibodies induced by a human parvovirus in a sample of body fluid, wherein said sample is subjected to an immunoassay, c h a r a c t e r i z e d in that an artificial antigen according to any one of claims 2 3nd 3 is used as a diagnostic antigen.
A method according.to claim 4, wherein said sample is subjected to enzyme-linked immunosorbent assay (ELISA) c h a r a c t e r i z e d in that an artificial antigen according to any one of claims 2 and 3 is used as a diagnos- tic coating antigen.
6. A diagnostic immunoassay kit for the detection of antibodies induced by a human parvovirus in a sample of body fluid, c h a r a c t e r i z e d in that it comprises as a diagnostic antigen an artificial antigen according to any one of claims 2 and 3.
7. A diagnostic immunoassay kit according to claim 6, c h a r a c t e r i z e d in that it additionall- comprises a positive standard serum sample, a negative standard serum sample, an enzyme conjugate, optionally a substrate for the enzyme conjugate, and optionally buffer solution(s) and/or washing solution(s).
8. A vaccine composition, c h a r a c t e r i z e d in that it comprises as an immunizing component, at least one antigen selected from the artificial antigens according to any one of claims 2 and 3, together with a nontoxic pharmaceutically acceptable carrier and/or diluent. $ijBLfV r E
9. An artificial peptide substantially as hereinbefore described with reference to any one of the examples. DATED: 2 February 1993 PHILLIPS ORMONDE FITZPATRICK Attorneys for: FERRING DIAGNOSTICA AB 14 INTERNATIONAL SEARCH REPORT International Application No PCT/SE 90/00276 I. CLASSIFICATION OF SUBJECT MATTER (if several classification symbols apply, indicate According to International Patent Classification (IPC) or to both National ClassificatiL,. and IPC C 07 K 7/10, G 01 N 33/569, A 61 K 39/23 :I i IELDS SEARCHED Minimum Documentatior Searched 7 Classification System Classification Symbols A 61 K, C 07 K, G 01 N Documentation Searched other than Minimum Documentation to the Extent that such Documents are Included in Fields Searched 8 SE,DK,FI,NO classes as above III. DOCUMENTS CONSIDERED TO BE RELEVANT 9 Category Citation of Document, 11 with indication, where appropriate, of the relevant passages 12 Relevant to Claim No. 13 Y EP, Al, 0117063 (AMGEN) 29 August 1984, 1 see in particular pages 7-8, 52 and 58-71 A 2-10 A EP, A2, 0238893 (ABBOTT LABORATORIES) 1-10 September 1987, see pages 11-12 A Journal of Virological Methods, Vol. 20, 1988 T.F. 1-10 Schwarz et al.: "Human parvovirus 819: ELISA and immunoblot assays see page 155 page 168 A Journal of Medical Virology, Vol. 23, 1987 J.P. 1-10 Clewley et al.: "Detection of Parvovirus 819 DNA, Antigen, and Particles in the Human Fetus see page 367 page 376 SSpecial categories of cited documents: 10 later document published after the international filing date document defining he enerai stale of the art wh'ich is not or pr"ority date and not in conflict with the application but considered tfbe ofparticular relevance aci w hte to understand the principle or theory underlying the considered be parcular relevance invention earlier document but published on or Iter the international filing date Xbu document of particular relevance, the claimed invention cannot be considered novel or cannot be considered to L document which may throw doubts pn priority claim(s) or involve an inventive step which is cited to establish the publication dale of another l citation or other special reason las specified) document of particular relevance, the claimed invention ctaion orcannot be considered to involve an inventive step when the 0 document referring to an oral disclosure, use, exhibition or document sh combin tiont bein orbvus to therson sc d ed o"ther mein. in the art. documeit published prior to the international filing date but document member of the same patent family later t'.an the priority date claimed document member of the same patent family IV. CERTIFICATION Date of the Actual Completion of the International Search Date of Mailing of this International Search Report July 1990 990 -07-2 International Searching Authority Sig 7 aure,of Aju.LrePnd Officer SWEDISH PATENT OFFICE rr iunF Form PCTIISAJ210 (second sheet) (January 1985) International Application No. PCT/SE 90/00276 IIl. DOCUMENTS CONSIDERED TO BE RELEVANT (CONTINUED FROM THE SECOND SHEET) Category- Citation of Document, with indication, where appropriate, of the relevant passages Relevant to Claim No J. gen. Virol., Vol. 68, 1987 G. Winkler et 1 al.: "Characterization of a Disulphide Bridge-stabilized Antigenic Domain of Tick-borne Encephalitis Virus Structural Glycoprotein see page 2239 page 2244 see in particular the abstract and page 2244 Journal of Virology, Vol. 61, No. 8, August 1987 1 John W. Gnann, Jr. et al.: "Fine Mapping of an Immunodominant Domain in the Transmembrane Glycoprotein of Human Immunodeficiency Virus see page 2639 page 2641 Form PCT/ISA/210 (extra sheet) (January 1985) ANNEX TO THE INTERNATIONAL SEARCH REPORT ON INTERNATIONAL PATENT APPLICATION NO.PCT/SE 90/00276 'h inex lists the patent family members relating to the patent documents cited in the above-mentioned international search report, members are as contained in the Swedish Patent Office EDP file on 90-07-04 The Swedish Patent Office is in no way liable for these particulars which are merely given for the purpose of information. Patent document Publication Patent family Publication cited in search report date member(s) date cD- 0117063 84-08-29 JP-T- 60500214 85-02-21 WO-A- 84/02847 84-08-02 EP-A2- 0238893 87-09-30 JP-A- 62224296 87-10-02
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SE8901566 | 1989-04-28 | ||
| SE8901566A SE8901566D0 (en) | 1989-04-28 | 1989-04-28 | NEW PEPTIDES INCLUDING A DISULFID BOND |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| AU5559690A AU5559690A (en) | 1990-11-29 |
| AU636145B2 true AU636145B2 (en) | 1993-04-22 |
Family
ID=20375840
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| AU55596/90A Expired - Fee Related AU636145B2 (en) | 1989-04-28 | 1990-04-25 | New human parvovirus peptides with disulfide bridge for immunization or diagnosis |
Country Status (6)
| Country | Link |
|---|---|
| EP (1) | EP0470205A1 (en) |
| AU (1) | AU636145B2 (en) |
| CA (1) | CA2053240A1 (en) |
| IL (1) | IL94218A0 (en) |
| SE (1) | SE8901566D0 (en) |
| WO (1) | WO1990013567A1 (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6274307B1 (en) * | 1990-02-08 | 2001-08-14 | Mikrogen Molekularbiologische Entwicklungs-Gmbh | Immunologically active peptides or polypeptides from the parvovirus B19 |
| US5244785A (en) * | 1991-02-01 | 1993-09-14 | Microgenics Corporation | Determination of high molecular weight analytes using a β-galactosidase complementation assay |
| JPH0910000A (en) | 1995-06-26 | 1997-01-14 | Nippon Sekijiyuujishiya | Method for detecting human parvovirus and reagent therefor |
| US6238860B1 (en) | 1998-11-05 | 2001-05-29 | Dyax Corp. | Binding moieties for human parvovirus B19 |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0117063A1 (en) * | 1983-01-19 | 1984-08-29 | Amgen | Methods and materials for development of parvovirus vaccine |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE3782350D1 (en) * | 1986-03-20 | 1992-12-03 | Abbott Lab | RECOMBINANT PARVOVIRUS RA-1 PRODUCTS AND METHOD FOR USE THEREOF. |
-
1989
- 1989-04-28 SE SE8901566A patent/SE8901566D0/en unknown
-
1990
- 1990-04-25 WO PCT/SE1990/000276 patent/WO1990013567A1/en not_active Ceased
- 1990-04-25 AU AU55596/90A patent/AU636145B2/en not_active Expired - Fee Related
- 1990-04-25 EP EP90908131A patent/EP0470205A1/en not_active Withdrawn
- 1990-04-25 CA CA002053240A patent/CA2053240A1/en not_active Abandoned
- 1990-04-26 IL IL94218A patent/IL94218A0/en unknown
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0117063A1 (en) * | 1983-01-19 | 1984-08-29 | Amgen | Methods and materials for development of parvovirus vaccine |
Also Published As
| Publication number | Publication date |
|---|---|
| IL94218A0 (en) | 1991-01-31 |
| SE8901566D0 (en) | 1989-04-28 |
| AU5559690A (en) | 1990-11-29 |
| WO1990013567A1 (en) | 1990-11-15 |
| CA2053240A1 (en) | 1990-10-29 |
| EP0470205A1 (en) | 1992-02-12 |
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