CN1214367A - Monoclonal antibody for human anti-hantaan virus glycoprotein neutralization gene engineering - Google Patents

Abstract

The present invention includes the Fab fragment antibody gene of the monoclonal antibody, gene product and its application, and it features that recombined antibody is specific recognition Hantaan virus glycoprotein, which is determined by the specific gene sequence existing in the hypervariable regions (CDRs) of the variable antibody gene light chain and heavy chain region, has obtained effective expression and has anti-virus neutralization activity. Its possible application is utilizing obtained neutralized Fab antibody gene and its expression product in the clinical treatment of hemorrhagic fever as kidney syndrome.

Description

People source anti-hantavirus glycoprotein neutrality gene engineering monoclonal antibody

The present invention relates to treat preparation and application, the especially specificity of personnel selection source gene engineering monoclonal antibody neutrality gene engineering monoclonal antibody at Hantaan virus glycoprotein.The utilization phage surface is expressed (PhageDisplay) technology, extracted total cell RNA in the periphery lymphocyte that from the convalescent's anticoagulation of popular hemorrhagic fever epidemic-stricken area, is separated to, by the RT-PCR method, with lineup IgGFab gene-specific primer, from synthetic cDNA pcr amplification one group of light chain and heavy chain Fab fragment gene, light chain and heavy chain are successively inserted phage vector pComb3, successfully set up anti-hantavirus antibody gene storehouse, and with the method that the Hantaan virus particle and the anti-hantavirus glycoprotein mouse monoclonal antibody of purifying are caught glycoprotein antigen this antibody library has been carried out rich long-pending screening and expressed, obtain people's source neutrality anti-hantavirus hantaan type gene engineering monoclonal antibody Fab fragment gene and expression thereof.Nucleotide sequence analysis confirms that the gene that obtains is the humanized IgG Fab gene, with specificity radioimmunoprecipitation (IP), IFAT and ELISA and PRNT are identified and are shown that human Fab's antibody recognition Hantaan virus glycoprotein g1 of expression has extracorporeal neutralizing activity.Use antibody drug, this neutralizing antibody to be expected to be used to the hemorrhagic fever with renal syndrome that Hantaan virus causes in the future as a kind of potential treatment.

Since B lymphocyte hybridoma cell-fusion techniques in 1975 comes out, monoclonal antibody is as clinical diagnosis, the researchdevelopment that the novel product of clinical treatment and prevention, its applying value and DEVELOPMENT PROSPECT have obtained general sure molecular biology and molecular immunology has caused the generation and the development of gene engineering monoclonal antibody.Reorganization by the antibody molecule gene level can obtain diversified specific murine source, rabbit source and human antibody, making has had breakthrough also more and more to demonstrate its significance and practice prospect both at home and abroad to gene engineering monoclonal antibody to studies on Monoclonal Antibody, especially year surplus the research of humanization or human antibody existing 10.And the phage antibody gene pool technology side that rise the beginning of the nineties at the end of the eighties has become the focus of world today's human source gene engineering monoclonal antibody research.The form that has developed at present successful gene engineering monoclonal antibody has: 1) people-mouse chimeric antibody, it is light to be about to the mouse monoclonal antibody, heavy chain variable region gene VL and VH are light with people's antibody respectively, weight chain constant area gene CL and CH are connected to form people-mouse chimeric light chain and heavy chain gene, and, obtain in insect cell and the Chinese hamster ovary celI to express the myeloma cell.2, reshaped antibody or humanized antibody promptly substitute the CDR district in people's monoclonal antibody Fab section or in people-mouse monoclonal antibody the decision of the surface in the FR district in mouse Fab section family base are replaced the corresponding gene order of adult's antibody with the CDR district, hypervariable region in the specific murine monoclonal antibody Fab section.China also has successfully report.3) small molecular antibody, comprise Fab (complete light chain and Fd), sFv (VH links to each other through a little connection peptides with VL), Fv (VH and VL form) VH and MRU (atom) obtain Fab or sFv and express in prokaryotic cell prokaryocyte by the phage antibody gene pool of hybridoma or foundation.4, the reorganization whole antibody, promptly whole immunoglobulin molecules comprises IgG, IgA is respectively at lymphoma cell, the expression in non-lymphoid cell such as Chinese hamster ovary celI and the rhabdovirus system.

Phage surface express polypeptide or albumen technology were originated in 1985.With this technique construction phage antibody gene pool is the focus of nearest 5-6 development.From the antibody expression form of antibody library screening mainly based on Fab section antibody and scFv.Made up at present and developed a lot in order to the phage vector system that makes up the antibody gene storehouse, wherein more representational expression system is the pComb system that is formed by the research of U.S. Scripps institute, its ultimate principle is that the repertoire antibody light chain gene group of pcr amplification and a complete set of heavy chain gene group are inserted among the phage vector pComb3, wherein Fd with the form of albumen III or albumen VIII fusion rotein, light chain is then with the independent form of expressing, be secreted into the bacterial cell gap and be assembled into Fab by leader pelB, after helper phage infects, form phage antibody library.Obtain the specific antibody gene by using specific antigens antagonist storehouse to carry out the long-pending screening of richness.Nineteen ninety Scripps institute obtains the human immunoglobulin gene storehouse first and obtains people's anti-tetanus toxin monoclone antibody from the donor peripheral blood B cell of tetanus toxin immunity.So far this technology is extensively with making up mouse and human immunoglobulin gene storehouse and make important progress, thereby is that the pCom3 expression system is set up antibody library and obtained antiviral human monoclonal antibodies and have successful report abroad by the phage vector expression system by it.The antiviral people's monoclonal antibody of having reported in the world at present that screens from phage human immunoglobulin gene storehouse has: the resisiting influenza virus monoclonal antibody, anti-HBsAg antibody, anti respiratory syncytial virus (RSV) F protein antibodies, anti-herpes simplex virus antibody, anti-HIVgp120 antibody and anti-hantavirus nucleoprotein antibody etc.Above-mentioned antibody all with Fab section form at expression in escherichia coli, and obtain the specific antibody gene order simultaneously.

Hantaan virus is the pathogenic agent that causes serious transmissible disease hemorrhagic fever with renal syndrome.Hemorrhagic fever with renal syndrome is widely current in China, and the patient about 100,000 is arranged every year, and its clinical case fatality rate is higher, serious harm people's life health, and causing the present does not have specific drugs treatment.The Hantaan virus gene is formed the nucleoprotein of encoding respectively, glycoprotein g1 and G2 and virus polymerase by S, M and three fragments of L.In the Hantaan virus and antigen mainly be present on the glycoprotein g1 and G2 by the M fragment coding.The neutralizing monoclonal antibody that experimental results show that anti-G1 and G2 has the passive protection effect that the passive protection suslik avoids the Hantaan virus attack.With epidemic hemorrhagic fever patient decubation anticoagulation or the clinical hemorrhagic fever patient of serum treatment, its curative effect obtains certainly in China epidemic-stricken area and external clinical application.Therefore the antibody preparation that the anti-hantavirus of a kind of standard of development infects under the situation of no specificity antivirus medicine has very important clinical meaning and good prospect.

Specific antiviral antibody can be used for the treatment of clinical disease viral disease and has obtained generally generally acknowledging.Weak points such as classical pathway exists difficulty big by cytogamy screening preparation people monoclonal antibody, and success ratio is low, and range of choice is little, and express cell system is unstable, and antibody titers is low.Set up human immunoglobulin gene year storehouse with molecular biology method, therefrom obtain stable specific antibody gene, can in protokaryon or eukaryotic cell, obtain the expression of efficient stable.And the gene that can improve the CDR zone on gene year level arbitrarily is to obtain the neutralizing antibody of high-affinity.Heavy chain, light chain make up the diversity that has increased the antibody that obtains greatly at random, can obtain people's monoclonal antibody that some are difficult to obtain with ordinary method.This is an important breakthrough in the human specific monoclonal antibody development technology.This investigative technique can promote the use the development in multiple antiviral monoclonal antibody, has far-reaching practice and be worth in the viral diseases treatment.

People-mouse chimeric antibody can be used for many-sided fundamental research, but as the clinical preparation of using, there is significant disadvantages in it.Though the Fc section of personnel selection has been replaced the Fc section of mouse, but the Fab section FR district of mouse still has heterology, can induce and produce anti-mouse body, lose some conformation of natural antibody and cause antibody activity decline or humanized antibody or reprovision antibody that specific murine monoclonal antibody CDR zone is substituted into the CDR district of people's antibody though be not suitable for using repeatedly some chimeric antibody, the CDR sequence in mouse source often and the FR sequence between it incompatible and cause the change of antibody conformation.Obviously, human source gene engineering monoclonal antibody is the optimal selection of development antiviral antibody.

The objective of the invention is by genetic engineering means and phage display technique in conjunction with utilization, directly from the human immunoglobulin gene storehouse, filter out the gene engineering monoclonal antibody of neutrality anti-hantavirus, obtain its antibody gene, for possible clinical antiviral therapy in the future provides the specific antibody medicine of feasibility.

The people source anti-hantavirus glycoprotein neutrality gene engineering monoclonal antibody Fab antibody of the present invention's statement comprises:

(1) is the reorganization IgG Fab antibody that in prokaryotic cell prokaryocyte, obtains to stablize effective expression in a kind of people source, called after AH23.

(2) its antibody protein function is by complementary region (the Complementarity-Dertemining Regions of decision family that is present in antibody gene light chain and variable region of heavy chain, what CDRs) the specificity nucleotide sequence determined among CDR1, CDR2 and the CDR3, its amino acid sequence corresponding has constituted the specific antigens calmodulin binding domain CaM (accompanying drawing 1) of antibody

(3) specific light chain and heavy chain gene derive from the rich long-pending screening expression of the specificity in anti-hantavirus antibody gene storehouse, people source, and corresponding three the CDR region sequences of light chain and heavy chain are the distinctive brand-new sequence of this antibody.

(4) specific recognition Hantaan virus envelope glycoprotein, and have the neutralization activity that anti-hantavirus infects

(5) utilize the neutrality Fab antibody gene of above-mentioned acquisition, contain any other gene of this antibody gene after can in prokaryotic cell prokaryocyte, yeast cell, eukaryotic cell and any recombinant virus system, expressing this antibody gene or reconstruction based on this, the antibody product that infects with Hantaan virus during acquisition has can be used for the treatment of the hemorrhagic fever with renal syndrome that is caused by Hantaan virus clinically.

Traditional hybridoma cell technology of utilizing obtains quite difficulty of human monoclonal antibody, and the clone of setting up is unstable usually, and gene is easily lost.The people source anti-hantavirus glycoprotein g1 neutrality gene engineering monoclonal antibody Fab antibody of the present invention's statement, it is the expression that on the basis that obtains antibody gene, obtains gene product, this antibody gene can be along with plasmid DNA duplicating and stable duplicating in bacterium, and can reconstruct antibody gene arbitrarily, thereby obtain a kind of clinical treatment antibody preparation of feasibility according to different needs.

Following preferential embodiment elaborates to the present invention, load does not mean that restriction content of the present invention in these embodiments, is explanation the present invention, and the employing phage expression vector is pComb3 (Barbas C. III etal, Proc.Natl.Acas.Sci USA 1991,89:10164-10168).Used main bacterial strain is commercial prod XLI-Blu (U.S. Strategene company product).Used phage is VCSM13 (U.S. Strategene company product).Used hantaan virus strain is international standard hantaan virus strain 76-118 (SschmaljohnC.S.et al, Science 1985,227:1041-1044) and the hantaan virus strain 84F-Li. of isolated in China (Liang M.F et al, Virus Research 1994,31:219-223).

Embodiment 1-4 is the screening preparation method of people source anti-hantavirus glycoprotein neutrality gene engineering monoclonal antibody Fab antibody A H23; Embodiment 6 is the gene expression characteristics of people source anti-hantavirus glycoprotein neutrality gene engineering monoclonal antibody FabAH23; Example 7-11 is albumen and the functional character of people source anti-hantavirus glycoprotein neutrality gene engineering monoclonal antibody Fab antibody A H23.

Example 1: the pcr amplification of humanized IgG Fab fragment gene: with lymphocyte separation medium (Shanghai chemical reagent factory) isolated lymphocytes from the patient's EHF decubation anticoagulation of epidemic-stricken area, Anhui; with Tril-Zon (U.S. Gibco; BRL) extract total cell RNA; with the Olig-dT primer RNA that extracts is passed through reversed transcriptive enzyme (U.S. Gibco; BRL) reverse transcription becomes cDNA; with a group-specific IgGFabGamma chain, Kampa chain and Lamda strand primer (table 1), people's endogenous light chain and heavy chain Fab gene are carried out pcr amplification.The PCR condition is: 94 ℃ 1 minute, 54 ℃ 1 minute, 72 ℃ 2 minutes, 35 circulations (PE 480), above-mentioned PCR product reclaims through sepharose respectively, through DNA purification column Spin-X (U.S. Gibco, BRL) behind the purifying, obtain Kamba, Lamda and Fd chain PCR product (accompanying drawing 2) about 650-700bp.

The foundation of example 2 phage antibody gene pools: different primer synthetic Kamba and Lamda chain PCR product are mixed, different primer synthetic Fd chains are mixed, be cloned into phage vector pComb3 with SacI/XbaI and XhoI/SpeI respectively, to be cloned into light, the pComb3 carrier DNA of heavy chain gene connects product behind ethanol sedimentation, hang with the 10ul aquae destillata, the 200ul electricity that it is good that prepared beforehand is gone in the electricity transduction changes bacterium XLI-Blu, (electric commentaries on classics condition is: Bio-Red electroporation, 0.2cm electricity revolving cup, 2.5K volt), electricity adds the 10mlSOC nutrient solution after changeing, 37 ℃ 1 hour, add the SB nutrient solution (3) that 10ml has penbritin and tsiklomitsin, 37 ℃ l hour, add the aforementioned SB of 80-100ml, 37 ℃ add helper phage M13GCM 1 * 10 12 after 2 hours, add kantlex (70ug/ml) after 1 hour, 37 ℃ of shaking table overnight incubation.With 4%PEG8000 and 3%NaCl precipitation phage supernatant, through 9000rpm, 20 minutes, 4 ℃ centrifugal after, with 2ml 0.02M PBS PH 7.4 resuspended precipitations, set up phage antibody library, packing is stored in-20 ℃ of refrigerator-freezers.

Example 3: be used for the antigenic preparation of Hantaan virus of antibody library enrichment screening: 1) Hantaan virus particulate purifying: according to literature method (10), the metainfective cell conditioned medium liquid of Hantaan virus 76-118 strain is precipitated with PEG, through the 10-60% sucrose density gradient centrifugation, receive the virion antigen of oyster white virus band as purifying.2) preparation of cells infected lysate: press literature method, use Hantaan virus 76-118,84-FLi infected behind the E-6 cell 7 days, used 1%Trito-100, and the lysate lysing cell is made Hantaan virus antigen.

Example 4: the rich long-pending screening of the specificity of phage antibody library: the Hantaan virus particle skimmed milk-PBS that uses above-mentioned purifying respectively, 37 ℃ of sealings add the every hole 60ul of above-mentioned phage antibody library after 2 hours, 37C was hatched 2 hours, use 5%Tween-20,-TBS washes 20 times repeatedly, use every hole 60ul at last, pH2.2 glycine one hydrochloric acid elutriant wash-out, the Tris liquid neutralization of pH9.6.Phage behind the wash-out continues to infect the XL1-Blu bacterium about the fresh OD600=1.0 of 2ml, carries out the next round screening after helper phage VCSM13 (U.S. Stratagene) infects.So repeated screening is 4-5 time, with every phage-infect XL1-Blu bacterium of taking turns after the specificity richness is amassed screening, gets an amount of bacterium (10-50ul) and is coated with 37 ℃ of ammonia Bian plates, after 4-5 hour, adds 5mMIPTG wetted nitrocellulose filter, and 30 ℃ are spent the night.This film with the sealing of 5% skimmed milk, adds the anti-human IgG Fab antibody staining of HRP mark after chloroform is fixing.Obtain long-pending enhancing of richness of specific antibody positive colony.As Fig. 3, be the long-pending screening of richness (nitrocellulose filter immunoblotting) in human immunoglobulin gene storehouse.

The nucleic acid sequence analysis of the variable region gene of example 5 human IgG Fab antibody A H23: the positive colony DNA with QiagenMiniprepKit (U.S. Qiagen) preparation is picked out, carry out nucleic acid sequence analysis with dideoxy method to specific people's anti-hantavirus IgGFab antibody variable gene.The order-checking used kit is Sequenase Version 2.0 Sequencing Kit (U.S. USB/Amacia).(5 ' CTAACTAGCTAGTCGCC) are used to measure chain variable region gene to derive from the primer of carrier pComb3; Heavy chain gene then needs to be gone into PCR II carrier (American I nvitro Gene) by subclone, measures with M13 or T7 primer then.At least 3 clones are used to determine same identical sequence.The sequence that obtains all uses DNAStrider (MS) sequence analysis software to carry out analyzing and processing, and compares the IgG sequence in the gene pool on the Internet network.The Fab gene of confirmer source anti-hantavirus glycoprotein neutrality gene engineering monoclonal antibody AH23 is made up of human IgG chain Fd and λ chain, its gene expression characteristics is made of the specificity nucleotide sequence and the amino acid in 6 CDR districts in VH and the VL structural domain, be VH-CDR1, VH-CDR2, VH-CDR3 and VL-CDR1, VL-CDR2 and VL-CDR3.As Fig. 1, be the Nucleotide and the aminoacid sequence of people source anti-hantavirus glycoprotein gene engineering monoclonal antibody Fab section antibody A H23 light chain and heavy chain gene variable region.

Example 6 will extract plasmid DNA after will having the positive colony amplification that AH23Fab antibody weight chain gene inserts according to a conventional method, with the g III in Spe I and the Npe I excision carrier, become Fdg III fusion rotein and be independent expressed proteins, connect the back and transform the XL1-Blu bacterium, the single bacterium colony of picking from the ammonia benzyl plate of overnight growth, inoculation SB or TB inoculum are when bacterium grows to OD=0.2-0.3, add 1mMIPTG, at 30 ℃ of abduction delivering 10-12 hours.Results bacterium, centrifugal back add 10 times of PBS (0.02M PH7.4) that concentrate amount of original fluid hangs, multigelation 3 times, and its supernatant is the Fab antibody of expression behind the high speed centrifugation.Hantaan virus is made the antigen sheet after infecting the Vero-E6 cell, add the Fab that expresses, the human source gene engineering monoclonal antibody AH23 that the anti-human IgG Fab antibody that adds fluorescein IFTC mark after 37Cf is hatched, specific immunity fluorescent reaction promptly confirm to obtain is at Hantaan virus.As Fig. 4, be the Immunofluorescence Reactions of the people source anti-hantavirus G1Fab antibody A H23 of expression to Hantaan virus.

Example 7 enzyme linked immunosorbent assay (ELISA): 1) use the Hantaan virus antigen 0.5ug of the purifying of preparation in the example 3 and anti-human IgG Fab antibody 0.1ug (U.S. Sigma) to wrap respectively by 96 orifice plates, the human Fab's expression product bacterial lysate that adds different dilution above-mentioned preparations, 37 ℃ hatch 1.5 hours after, add the goat anti-human igg Fab antibody of HRP mark. (Sigma).2) use anti-hantavirus nucleoprotein and glycoprotein monoclonal antibody bag by 96 orifice plates respectively, add the cell pyrolysis liquid that the Hantaan virus described in the aforesaid method 4 infects, further add anti-human Fab's antibody of human Fab's antibody and HRP mark.Chromogenic substrate is TMB, after the adding 4N sulfuric acid termination reaction, detects every hole OD value at OD450nm.The result confirms that the human source gene engineering monoclonal antibody AH23 of this acquisition is at Hantaan virus glycoprotein.As Fig. 5, be that enzyme linked immunoassay (ELISA) detects people source anti-hantavirus gene engineering monoclonal antibody Fab antibody.

Example 8 radioimmunoprecipitation specificity Hantaan virus structural protein: infect the Vero-E6 cell with excessive Hantaan virus 76-118, with 35S mark methionine(Met) and/or 3H labelled leucine labeled virus albumen, after the cells infected cracking of cell pyrolysis liquid after with mark, add the Fab solubility cracking supernatant liquor of expressing, the albumin A agarose that adds anti-human IgG Fab antibody coupling after 4 ℃ of night incubation, after washing, in precipitation, add the SDS-PAGE sample buffer, carry out the SDS-PAGE electrophoresis, specificity radioimmunity signal shows that the human source gene engineering monoclonal antibody AH23 that obtains is at the Hantaan virus glycoprotein g1.As Fig. 6, be that radioimmunoprecipitation is identified the specificity combination of the Fab of expression to the Hantaan virus structural protein.

Example 9 PRNTs (PRNT): in 24 porocyte culture plates, grow up to individual layer VeroE6 cell, to begin to carry out doubling dilution from stoste from the Fab solubility cracking supernatant liquor and the negative control of difference clone's expression, or the Fab and the bovine serum albumin (BSA) of purifying be normalized into 200ug/ml, mix with the Hantaan virus liquid of 30 PFU respectively then, hatched 1.5 hours for 37 ℃, every hole 100ul adds in the entering plate, each extent of dilution 2 hole, (100ml contains: 0.6 gram agarose to add the first layer 6% agarose then, 10% bovine serum, 1%PS, 1% non-essential amino acid, 4ml 7.5% glutamine and 2ml 1M Hepes).37 ℃, 5%CO ₂Hatched under the condition 7 days, and added second layer agarose (the same, 4% toluylene red) to the 8th day.With the high dilution of the antibody that reduces by 50% plaque number is NAT.The human source gene engineering monoclonal antibody AH23 that obtains have external in and the function that infects of Hantaan virus, be neutrality antibody.As Fig. 7, be that PRNT detects the neutralization activity of human Fab's antibody of purifying to Hantaan virus 76-118 and 84-Fli strain.

Example 10: the affinitive layer purification of human Fab's expression product: will resist the human IgG antibody to be coupled to ProteinA-Sepharose4B (U.S. Sigma), and carry out affinitive layer purification then.Obtain the electrophoresis pure protein of human source gene engineering monoclonal antibody AH23.As Fig. 8, be the SDS-PAGE of people source anti-hantavirus monoclonal antibody Fab section antibody A H23 expression product.Swimming lane is from left to right: 1.Mr: low molecular weight protein (LMWP) standard substance: 2. the AH23Fab 3. unpurified bacterial lysates that contain the AH23Fab expression product of purifying.

Claims (5)

1, people source anti-hantavirus glycoprotein neutrality gene engineering monoclonal antibody, it is characterized in that: (1) for its antibody protein function of people source anti-hantavirus glycoprotein neutrality gene engineering monoclonal antibody Fab antibody (2) of a kind of gene recombination that in prokaryotic cell prokaryocyte, obtains to stablize effective expression by the decision family complementary region that is present in antibody gene light chain and variable region of heavy chain (Complementarity-Dertemining Regions, CDRs) CDR1, the specificity nucleotide sequence determines among CDR2 and the CDR3, specific antigens calmodulin binding domain CaM (accompanying drawing 1) (3) specific light chain and heavy chain gene that its amino acid sequence corresponding has constituted antibody derive from the rich long-pending screening expression of the specificity in anti-hantavirus antibody gene storehouse, people source, and its corresponding three CDR region sequences are the distinctive brand-new sequence of this antibody.(4) specific recognition Hantaan virus envelope glycoprotein, and have the neutralization activity that anti-hantavirus infects

2, the purposes of people source anti-hantavirus glycoprotein neutrality gene engineering monoclonal antibody, it is characterized in that: the neutrality Fab antibody gene that utilizes above-mentioned acquisition, contain any other gene of this antibody gene after can in prokaryotic cell prokaryocyte, yeast cell, eukaryotic cell and any recombinant virus system, expressing this antibody gene or reconstruction based on this, the antibody product that infects with Hantaan virus during acquisition has can be used for the treatment of the renal syndrome-hemorrhagic fever that is caused by Hantaan virus clinically.

3, according to claim 1 people source anti-hantavirus glycoprotein neutrality gene engineering monoclonal antibody Fab antibody, it is characterized in that: the gene behaviour source anti-hantavirus neutrality antibody gene of the described specificity Fab antibody protein of encoding can be used to any expression system and express the anti-hantavirus neutralizing antibody.

4, according to claim 1 people source anti-hantavirus glycoprotein neutrality gene engineering monoclonal antibody Fab antibody, it is characterized in that: the gene behaviour source neutrality anti-hantavirus antibody gene of the described specificity Fab antibody protein of encoding, its nucleotide sequence or consequent aminoacid sequence can be used to the transformation of genes involved, and express the neutrality antibody or the polypeptide product of anti-hantavirus glycoprotein.

5, according to claim 1 people source anti-hantavirus glycoprotein neutrality gene engineering monoclonal antibody Fab antibody, it is characterized in that: during described antibody has on the identification Hantaan virus glycoprotein and antigen, thereby the function that the smooth poison of the blocking-up Chinese infects, its gene expression product is hoped and is used for the treatment of the hemorrhagic fever with renal syndrome that is caused by Hantaan virus clinically.

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