JPH08510635A - Hepatitis B escape mutant specific binding molecule - Google Patents

Abstract

  (57) [Summary] Molecules that specifically bind to the antigenic determinants of hepatitis B escape mutants include cell lines SMH HBs 145 / G / R / I (ECACC 92212312), SMH HBs 145 / R / I (ECACC 93052626), SMH HBs 145. / G / R / II (ECACC 93033109) or SMH HBs 145 / R / II (ECACC 93033110) secreted monoclonal antibodies and other specific binding molecules that cross-compete with these antibodies. The antibodies secreted by the cell lines SMH HBs 145 / G / R / I and SMH HBs 145 / G / R / II bind to mutant (escape mutant) HBsAG and wild type HBsAG. The antibodies secreted by the cell lines SMH HBs 145 / R / I and SMH HBs 145 / R / II bind the mutant but not the wild type.

Description

Detailed Description of the Invention       Hepatitis B escape mutant specific binding molecule   The present invention provides antibodies and other binding molecules specific for particular hepatitis B virus antigens. It is related. The present invention also relates to the diagnosis and treatment of molecules as described above. It is about use.   Infection with the hepatitis B virus (HBV) is a serious problem in various parts of the world. However, only a few vaccines are currently available. City against HBV Commercially available vaccines are usually hepatitis B virus in intact or recombinant form. Surface antigen (HBsAG). SmithKline Becom's product Ngerix-Be ™ (SmithKline Beecham product ENGERIX-B^TM) Is after It is an example of a person. The HBV antigen subtypes are serologically defined, and HBsAG Was found to be due to a single nucleotide change in the genomic region that encodes There is. However, until recently, all known subtypes of the antigen were α-antigens. It contains the determinant and is composed of amino acids 124 to 147 of HBsAg. Antibodies to α-antigenic determinants provide protection against all known subtypes Was obtained. In recent years, hepatitis B virus vaccine-guided escape mutants (va A ccine-induced escape mutant was reported. Carman et al., The Lancet, Volume 336, page 325. ~ 329 (1990)). This escape mutant is in the α-antigenic determinant Substitution mutation (substituti) from glycine to arginine at position 145 in It was shown to have a mutant protein of HBsAG including on mutant). W OA-9114703 relates to a variant HBsAg-based vaccine. And also discloses an antibody preparation comprising an anti-mutant HBsAg antibody. It   However, WO-A-9114703 describes how a mutant α-antibody is used. To get the specificity needed to bind only the original determinant region or what is the wild type And no suggestion that it binds to both the mutant α-antigenic determinant region Absent. The present invention addresses these twin problems.   According to the first concept of the present invention, it is possible to specifically bind to a hepatitis B antigenic determinant. Cell lines SMH HBs 145 / G / R / I (ECACC 921223 12), SMH HBs 145 / R / I (ECACC 93052626), SMH HBs 145 / G / R / II (ECACC 93033109) According to SMH HBs 145 / R / II (ECACC 93033110) Secreted monoclonal antibody or cross-compete with the above antibody (cross- compete) molecule.   A specific binding molecule, such as an antibody, may be conformationally linked as the same or otherwise. When connecting to a conformally linked location exactly, Cross to compete. Conformationally linked positions are adjacent to each other on the polypeptide chain of the antigen. At an adjacent location or due to the secondary structure of the polypeptide chain. So that non-adjacent regions fold adjacent to each other (adja cent folding). Cross-competition experiments can be performed relatively easily (water (Waters) et al., Virus Research, Vol. 22, page. 1-12 (1991)), the target antibody or other specific binding molecule Determine whether specific cross-competition with monoclonal antibodies occurs as described above It is a clear method of   At least a portion cross-competes with a particular monoclonal antibody (ie, from 0% Specific binding molecules (having a significantly greater degree of cross-competition) are used in the present invention . All cross-compete (ie, have a degree of cross-competition that is not significantly less than 100%) Specific binding molecules are preferred, at least in some cases.   The specific binding molecule that can be used in the present invention is often an antibody itself. Yes. Polyclonal antibodies can also be used, but because the specificity is much more accurate, Monoclonal antibodies are usually preferred. Monoclonal antibody technology is based on the hler) and Milstein (Nature (N ature), vol. 256, page 495 (1975)) It's been fairly well established, and today it routinely produces monoclonal antibodies. There are many protocols used for. For example, a technology that can be used is Gefter et al. (Somatic Cell G enetics), Volume 3, page 231 (1977)), Kehler et al. Euro J. Immunovirol., Pages 292-2. 95 (1976) and Goding (“monoclonal antibodies”). Diseases: Principles and Practices (Monoclonal Antibodies: Pri nciple and Practice) "(2nd edition, 1986), Academic Press (Aca demic Press), New York). In particular, the following protocols Use le:   (A) A test animal (mouse, etc.) is placed at the antibody (in this case, the 145th position Immunization with an antigen that produces HBsAg) containing a substitution mutation from syn to arginine Place (immunologically challenge);   (B) Next, the splenocytes of the animal were fused with cells of the myeloma cell line, and the resulting hive Spread the fused cells as a redoma in selective medium;   (C) screening for specific antibodies by any suitable technique, eg, from another species By using an anti-immunoglobulin antibody from   As a general rule, the use of human monoclonal antibodies is also intended, especially for the treatment of humans. And in vivo diagnosis, but it is technically difficult. Lidoma technology is not suitable for producing large numbers of human monoclonal antibodies . Therefore, non-human monoclonal antibodies such as those of mouse origin are actually used. Often.   Chimeric antibodies, especially chimeric monoclonal antibodies, are also included in the concept of the present invention. . Such chimeric antibodies include SMH HBs 145 / G / R / I, SM H HBs 145 / R / I, SMH HBs 145 / G / R / II or S Sufficient to have its characteristic specificity from MH HBs 145 / R / II An amino acid sequence is mentioned. At a minimum, the complementarity-determining regions of the antibody of interest should be It is present to a sufficient extent to maintain the opposite sex. All V_HAnd V_LDomain exists , Or Fab or F (ab ') that is enzymatically induced₂Complete anti-fragment It means that a body-binding fragment is present.   There are various techniques for preparing chimeric antibodies. For example, rodent V A chimeric antibody composed of a human C region fused to the region has been reported (Mori). Morrison et al., PNAS, Volume 81, pages 6851-6855 ( 1984), Boulianne et al., Nature, Volume 312 , Pages 643-646 (1984) and Neuberger et al., Nature ure), volume 314, pages 268-270 (1985)). Or chimeric anti Body technology is also described in WO-A-9004413 and WO-A-9116354. Listed, these are antibody conjugates with two or more covalently linked Fc regions ( conjugate).   Fully humanised antibodies, especially monoclonal antibodies, are also available. Included in the concept of invention. It is common for non-human (especially mouse) antibodies to be adapted to the human body. There are three methods. Reichmann et al. (Nature, 332 Vol. Pp. 323-327 (1988)) suddenly reveals a specific site on ssDNA. It used mutagenesis (site-directed mutagenesis). As an alternative Jones, et al. (Nature, Volume 321, page 52). 2-525 (1986)) and Queen et al. (PNAS), 8 Volume 6, pages 10029-10033 (1989)) is a human framework. The complementarity-determining region of the rodent (C The entire V region was constructed using overlapping oligonucleotides with (DR) insertion. further Recently, Lewis and Crowe (Gene, Volume 101, Pages 297-302 (1991)) describes the polymerase chain reaction (PCR). Used to insert rodent CDRs onto the framework of human immunoglobulin ( graft). WO-A-9316192 is generally applicable to the human body for hepatitis. Was The present invention relates to an antibody and has been adapted to the human body according to the present invention by the suggestion of the above publication. Antibodies can be produced.   Cloning of heavy and light chain variable region amino acid sequences of monoclonal antibody Determined from the complemented DNA, the hypervariable region (or the complementarity determining region ... C DR) Kabat et al. (“Sequences of Proteins of   Immunological Interest (Sequences of Proteins of Immunological I nterest) ”, US Department of Health and Human   Services (US Department of Health and Human Services), USGA Valent Printing Office (US Government Printing Office), 1 987). Using any of the above methods, Inserted in human framework.   Ward et al. (Nature, Volume 341, pages 544-546) (1989)) single domain antibody (dAb ) Is a different class of specific binding molecule (whether it is appropriate to consider it as an “antibody”) Are shown separately) and are also included in the concept of the present invention. According to the above approach Then, using PCR or other suitable technique, V_HOr V_LClonin the gene And express it in a heterologous host such as E. coli.   The variable regions of the heavy and light chains are hybridized using the polymerase chain reaction (PCR). Expression vector amplified from dorma Clone in. The isolated variable region is screened by binding to the antigen. And its affinity is determined. Other single domain antibodies are immunized Amplified by the rearranged variable region gene in the spleen DNA of the isolated mouse Obtained directly by. After the amplified DNA has been cloned into the vector , Screened for antigen binding activity. Bacterio as an expression vector It has a variable gene because it is expressed on the cell surface by purification using phage. Phage can be directly selected by antigen (McCafferty et al. , Nature, 348, pages 552-554 (1990)).   dAb technology is a method by which recombinant DNA methods have specific binding ability. To completely change the occurrence of. If there is no other reason, As is generally understood, the present invention provides (polyclonal or monoclonal It should not be understood to be limited to antibodies (whether or not). Usually in the present invention For engineered engineered antibodies that can be used, see Winter and Milstein, Nature, Volume 349, pages 293- 299 (1991) and Marks et al., J. Baiorchem (J ． Biol. Chem.) 267, pp. 16007-16010, and 19 Successful X-Prostitution, London, March 7-8, 1994 Research of Biomedical Research (S Medical Lisa at Successful Exploitation of Biomedical Research Dr. Greg at the Medical Research Counsil Conference See the presentation by Dr. Greg Winter.   Specific binding molecules included in the concept of the present invention are related to mutant HBsAg binding ability. As far as I am doing, I can roughly divide it into two classes. First, mutant HBs It is a molecule that binds only to Ag and does not bind to wild-type HBsAg. Cell line SMH   Secreted by HBs 145 / R / I and SMH HBs145 / R / II These monoclonal antibodies, as well as specific binding molecules that cross-compete with them, Belongs to the category of. Secondly, the mutated HBsAg and wild type HBsAg It is a specific binding molecule that binds to both. Sometimes these specific binding molecules Is preferred. Specifically, SMH HBs 145 / G / R / I and SMH H Monoclonal antibodies secreted by Bs 145 / G / R / II and these Specific binding molecules capable of cross-competing with   According to a second concept of the invention, expressing a specific binding molecule as described above, preferably Provides a cell line or cell culture that can be secreted.   Hybridoma cell lines as detailed above are included within the concept of the invention. These cell lines are based on the European Collection of Animal Cell Chars (Eur opean Collection of Animal Cell Cultures) (ECACC) S Center for Applied Microbiology and Resar Chi (PHLS Center for Applied Microbiology and Research), Division Division of Biologics, Porton Down, Sari Subarry, Wiltshire SP 40 Jaze (Porton Down, Sal isbury, Wiltshire SP4 0JG), United Kingdom) Deposited on date of deposit:   These deposits were made under the Budapest Treaty.   General methods for making other monoclonal antibodies have been described above. These monochrome To confirm that null antibodies are included in the concept of the present invention, a simple cross-competition experiment was performed. The test was deposited The reaction may be performed using the antibody secreted by the cell line. Included in the concept of the invention Other specific binding molecules have been suggested in the literature and / or patent publications cited above. Can be manufactured.   Specific binding molecules according to the invention are useful in diagnosis and in therapy .   When used for diagnosis, the present invention, as one embodiment, is different from conventional hepatitis B. It can be used for the purpose of identifiable diagnosis of re-mutant hepatitis B. In this case, SM H HBs 145 / R / I or SMH HBs 145 / R / II Or use a specific binding molecule with this specificity as it binds only to the variant. Can be The specific binding molecule of the present invention binds only to a mutant hepatitis B antigen. See above with antibodies or other specific binding molecules that do not bind to wild type. EP-A- is a preferred monoclonal antibody. There is RF HBs 1 disclosed in 0038642.   In another embodiment, by using the present invention, wild type or The presence of mutant hepatitis B can be detected. In this case, SMH HBs 145 / G / R / I or SMH HBs 145 / G / R / II or this Use because specific binding molecules with specificity bind both wild type and mutant Can be   The diagnostic method according to the present invention is performed in vitro. According to a third concept of the invention, it is suspected that it contains hepatitis B particles or antigens. Of hepatitis B, which comprises contacting the sample in contact with the specific binding molecule described above. It provides a diagnostic method.   There are many types of in vitro assay methods. Labeled specific such as antibody While sometimes due to the use of a binding molecule, which use is within the concept of the invention, Antibodies (or other specific binding molecules) and antigens by observing the resulting precipitate In some cases, by detecting the interaction of. Qualitative and independent of labeled antibody And quantitative in vitro assays include, for example, gel precipitation Double diffusion method of Kuterronie, etc.), simple radial immunodiffusion method (SRID), rocket exemption Immunoelectrophoresis such as epidemiological electrophoresis and two-dimensional immunoelectrophoresis, and scattering of incident light source Quantification (nephelometric analysis).   In practice, it is often the case that some form of label is used to detect antigen-antibody interactions. Sometimes used to get out. The label may or may not be radioactive . Depending on the type of assay, specific binding molecules within the concept of the invention may be labeled with Or other specific binding that binds to the specific binding molecule included in the concept of the invention. The molecule may be labeled. Immunoassay (including radioimmunoassay) and immunoassay Assay (immunometric radioassay) (Including enzyme-linked immunosorbent assay) Can be used, and also the immunoblotting technique You may use. Labeling with chemiluminescent substances and labeling with fluorescent dyes are also included.   In vitro assays are often performed with kits. Fourth outline of the present invention As a reminder, the specific binding molecule described above and the specific binding molecule are hepatitis B particles. Alternatively, hepatitis B particles or a hepatitis B particle comprising means for detecting whether or not the antigen is bound. Or an assay kit for detecting an antigen.   This assay may be used, for example, in any of the above assays. Competition Competitive kit and especially the sandwich immunoassay kit (s andwitch immunoassay kit) is preferred. Specific binding molecule and detection means You can put it in another room in the room. Specific binding molecule bound to solid support It may have been done. The detection means is actually a second detectably labeled second. It preferably comprises a specific binding molecule (which itself may be an antibody). A second specific binding molecule of the above binds to the antibody or other specific binding molecule described above.   The present invention is also applicable to in vivo diagnosis. According to the fifth concept of the present invention And the specific consequences described above in the preparation of a medicament for the in vivo diagnosis of hepatitis B. It provides for the use of synthetic molecules (usually labeled). Therefore, The light indicates the specific binding component labeled above if necessary. Hepatitis B, which comprises administering the offspring to the patient, generally by parenteral means The present invention relates to a method for in vivo diagnosis. Labels that can be used in vivo include Radiolabels and paramagnetic labels are included, which can be detected by suitable external equipment. Wear.   In addition to being applicable to diagnosis, the present invention provides therapy, in particular treatment of hepatitis B infections, Can be used for passive immunization against hepatitis B.   According to a sixth aspect of the present invention, there is provided a therapeutic or prophylactic agent for hepatitis B infection. It provides the use of the above specific binding molecules in the preparation. Therefore, the book The invention is to administer the above specific binding molecules to a patient, generally parenterally. It can be used in a method for treating or preventing hepatitis B infection.   According to a seventh concept of the present invention, the above-mentioned specific binding molecule and pharmaceutically It is intended to provide a formulation of carriers which can be used.   Formulations administered parenterally to humans are usually sterile. Water for injection A carrier is present, such as an acid buffer solution. Dosage and timing n) or the specifics of the specific binding molecule to be administered and determined under the direction of the physician. Determined not only by isomerism and binding activity but also by antigenicity and other unfavorable characteristics Can be However, in general, each regimen for specific prevention is 1 to 1 a week 0 mg (for example, about 5 mg) may be mentioned as a specific therapeutic regimen. Therefore, 0.5 to 5 mg (for example, about 1 mg) is administered daily for about 1 week, and then 0 every week. ． It is envisaged to administer 5 to 5 mg (eg about 1 mg).   The preferred embodiment of each concept of the present invention is (mutatis mutand) with some modifications. is) Similar to other concepts.   The invention is illustrated by the examples below.Example 1-Preparation of hybridoma antigen   Yeast strains as described in Example 2 of WO-A-9114703. DC5 cir ° was transformed with plasmid pRIT13557 and transformed into Y1648. A strain was created. In the Y1648 strain, the 145th position was mutated from Gly to Arg. It also expresses mutant HBsAg. Mutant HBsAg was added to Aerosil (trademark) ( AEROSIL^TM) Adsorption / desorption, ultrafiltration, ion-exchange chromatography, sodium chloride Density gradient centrifugation with cesium (CsCl) and gradient fractions with cesium chloride (CsCl) By dialysis of Cl gradient fraction, the culture of Y1648 strain (with C1334 Named)). The batch of purified antigen was designated 31M5.Immunization   Inbreeding Balb / c mouse (Harlan Olac Limited) (Harlan Orac Ltd), Vicestar, Oxon (Bicester, Oxon, UK), 100 μl complete Freund's horse mackerel 40 μg recombinant mutant HBsAg from batch 31M5 emulsified in tubant Was subcutaneously administered and immunized. After 4 weeks, 100 μl of incomplete Freund's Ajuba 20 μg of a second dose of 31M5 emulsified with a detergent was administered subcutaneously. 2 weeks Then, 10 μg of 31M5 was intravenously administered to the mouse. 4 weeks later, fusion I killed this mouse for the purpose.fusion   Each splenocyte was washed and the number of cells was counted. These cells are the mouse myeloma cells System P3-NS-1 / 1-Ag4-1 (Flow Laboratories Limited (Fl ow Laboratories limited), Irvine, Scotland) and 10 Fused at a ratio of 1: 1. Polyethylene glycol 1500 as a fusion inducer Using. Fusion was continued for 7 minutes at 37 ° C. 2 x 10 fused cells⁶/ 2m One well was seeded and hybridomas were selected using HAT medium.Example 2-Screening for specific antibodies   The supernatant of the hybridoma-containing wells obtained in Example 1 was used as a recombinant wild type enzyme.   Ay sub-type HBsAG or 31M5 (mutant HBsAG) incubated with polystyrene beads coated with either sAg) . Antibodies bound to beads, horseradish peroxidase-labeled rabbit anti-mouse Immunoglobulin (Sigma Chemi Detected by Cal Company Limited (Sigma Chemical Company limited) did.Example 3-Cloning and Deposit   All wells tested by limiting dilution three times or for specific antibody Clone selected wells until positive for A body cell line was obtained. Specific for both mutant and wild type HBsAg A two-cell line secreting a monoclonal antibody European Collection of Animal Cell C ultures) (Porton Down, Salisbury, Wilt Shire (Porton Do wn, Salisbury, Wiltshire)) with the date and deposit number shown in the table below. Was:   Binding of SMH HBs 145 / G / R / I to mutant and wild type antigens Measured by enzyme-linked immunosorbent assay (ELISA) as follows: Decided The solid phase is coated with either mutant or wild type antigen and Incubated for 6 hours. Then wash this solid phase After cleaning, the horseradish peroxidase-labeled rabbit anti-maize for 2.5 hours at 37 ° C. Incubated with 1000 dilutions of Us antibody (Dako). this Wash the solid phase again and apply the enzyme substrate to the solid phase in a clean tube for 30 minutes. added. Read the OD reading of this liquid at 495 nm in a multiwell It measured with the plate reader (multi-well plate reader). This value is Control and improper IgG monoclonal antibody control Compared to other assays. The results are shown in the table below. OD reading for 2 antigens , Was significantly larger than the control reading (data not shown).   Identical binding of SMH HBs 145 / G / R / II to mutant and wild type antigens It was measured in the same manner.   Secretes a monoclonal antibody with specificity only for mutant HBsAg A two-cell system using the European Collection of Animal Cell Culture (European Collection of Animal Cell Cultures) (Portunda Eun, Salisbury, Wiltshir (Porton Down, Salisbury, Wiltshir e)) was deposited with the date and deposit number shown in the table below:   SMH HBs 145 / R / I and SMH H to variant and wild type antigens The binding of Bs 145 / R / II is described in SMH HBs 145 / G / R / I. The measurement was performed in the same manner as described above. In a similar manner, there was no significant It was shown that there was no binding.Example 4-Use of a monoclonal antibody to detect both mutant and wild type antigens   SMH HBs 145 / G / R / I or SMH HBs 145 / G / R Type or mutant type in serum sample containing antibody secreted by / II Detection of the virus in Escherichia coli using a 2-site immunoassay Goodal et al., Med. Lab. Sci., Volume 38, Pages 349-354 (1981)). That is, the antibody is captured at the capture phase. as) Coat on a working solid support. This is the time for which the optimal binding is obtained. Incubate with The sample is washed from the solid support. Antigen binding The amount is measured by incubating with labeled antibody. Protein Protein A affinity chromatography Antibodies purified from ascites by detecting radionuclide enzymes or eg biotin Labeled with another molecule that can The concentration of labeled antibody used in the assay and The incubation time and temperature are adjusted for optimal binding. Occlusion The shear protein is added to the antibody at this stage, for example fetal bovine serum or bovine serum albumin. Non-specific binding of proteins. Excess labeled antibody is washed away and bound label is removed. Sense is detected using an appropriate method. For example, biotin-labeled antibody can be And horseradish peroxidase complex. The binding of this complex Detected by ortho-phenylenediamine and hydrogen peroxide; light of the above color change The optical density is measured at 492 nm. Each assay included positive and negative controls. Mu.Example 5-Use of a monoclonal antibody to detect only variant antigens   Detection of mutant viruses only in serum samples containing antibodies was performed using SMH HB s 145 / G / R / I or SMH HBs 145 / G / R / II SMH HBs 145 / R / I or SM instead of secreted antibody Examples, except using antibody secreted by H HBs 145 / R / II The procedure was as described in 4.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁶ Identification code Internal reference number FI G01N 33/576 9162-4B C12N 15/00 C 33/577 9281-4B 5/00 B (81) Designated country EP (AT, BE, CH, DE, DK, ES, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OA (BF, BJ, CF, CG, CI, CM, GA , GN, ML, MR, NE, SN, TD, TG), AU, BB, BG, BR, BY, CA, CN, CZ, FI, GE, HU, JP, KG, KP, KR, KZ, LK. , LV, MD, MG, MN, MW, NO, NZ, PL, RO, RU, SD, SE, SI, SK, TJ, TT, UA, US, UZ, VN (72) Inventor Kerman, Wei Liam, Fredrick United Kingdom, Glasgow 42 42 Knu, Queens Park, Queens Drive 148 (72) Inventor Thomas, Howard, Christopher United Kingdom, London 29 NX, Beach Drive 39

Claims (1)

[Claims] 1. Cell line SMH HB capable of specifically binding to antigenic determinants of hepatitis B s 145 / G / R / I (ECACC 92122312), SMH HBs 145 / R / I (ECACC 93052626), SMH HBs 145 / G / R / II (ECACC 93033109) or SMH HBs 145 Secreted by / R / II (ECACC 93033110) Molecules that are or cross-compete with these antibodies. 2. The molecule according to claim 1, which is an antibody. 3. The molecule according to claim 2, which is a monoclonal antibody. 4. 4. The antibody according to claim 3, which is an antibody adapted to human or at least a part of human body. The molecule described in. 5. The non-human CDR is inserted on the framework of human immunoglobulin. 5. The molecule according to item 4 in the range. 6. A single domain antibody according to any one of claims 1 to 5. Molecule. 7. Binding to mutant HBsAg but not wild-type HBsAg The molecule according to any one of claims 1 to 6. 8. Cell line SMH HBs 145 / R / I (ECACC 93052626) Or SMH HBs 145 / R / II (ECACC 93033110) secreted monoclonal antibody body. 9. A method of binding to both mutant HBsAg and wild type HBsAg. The molecule according to any one of items 1 to 6. 10. Cell line SMH HBs 145 / G / R / I (ECACC 921223 12) or SMH HBs 145 / G / R / II (ECACC 93033 109) which is a secreted monoclonal antibody. 11. Expression of the specific binding molecule according to any one of claims 1 to 10. A cell line or cell culture capable of being. 12. The cell line according to claim 11, which is a hybridoma. 13. Cell line SMH HBs 145 / G / R / I (ECACC 921223 12). 14. Cell line SMH HBs 145 / G / R / II (ECACC 93033 109). 15. Cell line SMH HBs 145 / R / I (ECACC 93052626 ). 16. Cell line SMH HBs 145 / R / II (ECACC 9303311 0). 17． Use of any one of claims 1 to 10 for diagnosis and / or treatment A specific binding molecule described therein. 18. Claim a sample suspected of containing hepatitis B particles or antigen The step of contacting with the specific binding molecule according to any one of items 1 to 10 A method for diagnosing hepatitis B. 19. A specific binding molecule according to any one of claims 1 to 10 and A procedure for detecting whether or not this specific binding molecule binds to hepatitis B particles or antigen. An assay kit for detecting hepatitis B particles or antigens, which comprises a step. 20. Claim 1 in the preparation of a medicament for the in vivo diagnosis of hepatitis B. Use of the specific binding molecule according to any one of items 1 to 10. 21. A patient is provided with the specific binding molecule according to any one of claims 1 to 10. In vivo diagnostic method for hepatitis B, which comprises administering to 22. Claims in the preparation of a therapeutic or preventive drug for hepatitis B infection Use of the specific binding molecule according to any one of items 1 to 10. 23. A patient is provided with the specific binding molecule according to any one of claims 1 to 10. A method for treating or preventing hepatitis B infection, which comprises administering to 24. A specific binding molecule according to any one of claims 1 to 10 and A formulation comprising a pharmaceutically usable carrier.