CN102462838A - System for rapidly, efficiently and industrially producing biological vaccine - Google Patents
System for rapidly, efficiently and industrially producing biological vaccine Download PDFInfo
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- CN102462838A CN102462838A CN2010105454657A CN201010545465A CN102462838A CN 102462838 A CN102462838 A CN 102462838A CN 2010105454657 A CN2010105454657 A CN 2010105454657A CN 201010545465 A CN201010545465 A CN 201010545465A CN 102462838 A CN102462838 A CN 102462838A
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Abstract
The invention discloses a system for rapidly and efficiently producing a biological vaccine and a biological vaccine produced by using the same. In the system for producing the biological vaccine, an antigenic protein or an antigenic protein fragment is decided by at least one microorganism cell, at least one expression vector and at least one antigenic nucleotide sequence to produce the biological vaccine. The biological vaccine generated by using the system for producing the biological vaccine can be applied to human beings or animals. The animals can include poultry, birds, livestock and other mammals. The system for producing the biological vaccine has the advantages of rapidness and high efficiency, and has wide suitability. The biological vaccine generated by the system for producing the biological vaccine has high stability and a long preserving period.
Description
Technical field
The invention belongs to biotechnology and botany field; More specifically, the present invention relates to a kind of biovaccine production system and biovaccine of being produced thereof rapidly and efficiently.
Background technology
Vaccine plays an important role in infectious diseases such as control human and animal diseases, particularly human and animal's influenza.Present existing industrialization production of vaccine system has long, limitation such as the vaccine kind is limited of production cycle.Simultaneously, the vaccine that existing industrialization production of vaccine system produces mostly must be difficult to carry out extensive human and animal's vaccine and throw in via injecting pathway.
Therefore, be sought after a kind of industrial biological production of vaccine system rapidly and efficiently at present.
Summary of the invention
The object of the present invention is to provide a kind of biovaccine production system and biovaccine of being produced thereof rapidly and efficiently.
In first aspect of the present invention, a biovaccine production system is provided, described system comprises:
(A) at least a microbial cell; Described microbial cell is anthrax bacillus cell (Bacillus anthracis) or nontoxic anthrax bacillus cell (Bacillus anthracis Sterne strain); Its defective or disappearance are selected from following gene: anthrax bacillus lethal factor gene (LF); One of anthrax bacillus virulence factor gene (PI-PLC), anthrax bacillus edema factor (EF), or its combination;
(B) at least a construction, described construction comprises: at least a antigen nucleotide sequence, described antigen nucleotide sequence is expressed at least a antigen protein or antigen protein fragment.
In a preference, (B) in, described construction is an expression vector, this expression vector can exist in described microbial cell or duplicate;
And described antigen nucleotide sequence is integrated in this expression vector and can in described microbial cell, expresses and produces described antigen protein or antigen protein fragment.
In another preference, described antigen protein or antigen protein fragment are present in (being expressed in) described microbial cell, cell surface or extracellular.
In another preference, in described anthrax bacillus cell or the nontoxic anthrax bacillus cell:
(a) comprise the nucleotide sequence of anthrax bacillus lethal factor gene (LF), but the 331st to 418 disappearance of the aminoacid sequence of the anthrax bacillus lethal factor of its coding;
(b) do not comprise the nucleotide sequence of anthrax bacillus lethal factor gene (LF)
(c) do not comprise the nucleotide sequence of anthrax bacillus virulence factor gene (PI-PLC);
(d) comprise the nucleotide sequence of anthrax bacillus anthrax bacillus edema factor gene (EF), but the 359th to 601 disappearance of the aminoacid sequence of the anthrax bacillus edema factor of its coding; Or
(e) above (a)-(d) binomial or above combination of binomial arbitrarily.
In another preference, described construction can be integrated on described anthrax bacillus cell (Bacillus anthracis) or nontoxic anthrax bacillus cell (Bacillus anthracis Sterne strain) genome or the plasmid.
In another preference, described construction (like expression vector) can be integrated in the lef gene.
In another preference, described antigen protein or antigen protein fragment derive from one or more microorganisms that are selected from down group: adenovirus (Adenovirus), cytomegalovirus (Cytomegalovirus); Dengue virus (Dengue virus), nerpes vinrus hominis's the 4th type 10 (Epstein Barr virus), HAV (Hepatitis A); Herpes simplex virus (Herpes virus), influenza A virus (Influenza A virus), influenza A virus (Influenza A H1N1); Influenza B virus (Influenza B virus), Measles virus (Measles virus), mumps virus (Mumps virus); Parainfluenza virus 1 (Parainfluenza1), parainfluenza virus 2 (Parainfluenza 2), parainfluenza virus 3 (Parainfluenza 3); Rotavirus (Rotavirus), foot and mouth disease virus, respiratory syncytial virus (Respiratory Syncytialvirus); Rubella virus KOS (Rubella KOS), rubella virus K1S (Rubella K1S), rubella virus K2S (Rubella K2S); Chickenpox one varicella zoster virus (Varicella-Zoster virus), mycoplasma pneumoniae (Mycoplasma pneumoniae), chlamydia trachomatis (Chlamydia trachomatis); CPN (Chlamydia pneumoniae), or its combination.
In another preference, described antigen nucleic acid sequence encoding part or total length influenza virus HA (influenza hemagglutinin).
In another aspect of this invention, a kind of biovaccine is provided, produces by described biovaccine production system.
In a preference, described biovaccine is:
Described anthrax bacillus cell or nontoxic anthrax bacillus cell, it contains any one or a plurality of above-mentioned antigen protein or antigen protein fragment; Or
The spore of described anthrax bacillus cell or nontoxic anthrax bacillus cell, it contains any one or a plurality of above-mentioned antigen protein or antigen protein fragment.
In another aspect of this invention, the purposes of a kind of biovaccine in preparing the medicine that prevents, controls or treat following disease, disease or situation is provided: flu, fluidity flu, foot and mouth disease, pneumonia, hepatitis, tracheitis, diarrhoea, herpes, endophthalmitis, keratitis, measles, parotitis, German measles and varicella-zoster.
In a preference, the route of administration of said medicine is following any: oral, suck, and the Sublingual absorbs, buccal absorption, nasal absorption, urethra absorbs and anal absorbs.
Others of the present invention are because the disclosure of this paper is conspicuous to those skilled in the art.
Description of drawings
A sample of Fig. 1, biovaccine production system of the present invention.Express vector integration in this sample on cellular genome.
Wherein, 1 expression cell, 2 expression expression vectors, 3 expression antigen nucleotide sequences, 4 expression cellular genome, 5 expression antigen protein or antigen protein fragments, the spore of 10 expression anthrax bacillus cells or nontoxic anthrax bacillus cell.
Another sample of Fig. 2, biovaccine production system of the present invention.Expression vector is that plasmid is present in the cell in this sample.
Wherein, 1 expression cell, 2 expression expression vectors, 3 expression antigen nucleotide sequences, 4 expression cellular genome, 5 expression antigen protein or antigen protein fragments, the spore of 10 expression anthrax bacillus cells or nontoxic anthrax bacillus cell.
Fig. 3, can be used for a sample of the expression vector of biovaccine production system of the present invention.
(A) plasmid pSK108 has the whole anthrax bacillus lef gene that comes from the pXO1 plasmid and comprises～nucleotide sequence that the 900bp upstream and downstream comes from the pXO1 plasmid.Cloning vehicle is pBluescript II.
(B) plasmid pSB1 comprises whole anthrax bacillus lef gene nucleic acid sequence of coming from the pXO1 plasmid.Cloning vehicle is pUC19.(C) plasmid plefHa1 has influenza virus A (influenza A/Puerto Rico/8/34) hemagglutinin (HA) (influenza HA) nucleotide sequence that obtains through PCR from plasmid pT3PR8.
The specific embodiment
The inventor provides a biovaccine production system through deep research, and this system comprises:
(A) at least a microbial cell; Described microbial cell is anthrax bacillus cell (Bacillus anthracis) or nontoxic anthrax bacillus cell (Bacillus anthracis Sterne strain); Its defective or disappearance are selected from following gene: anthrax bacillus lethal factor gene (LF); One of anthrax bacillus virulence factor gene (PI-PLC), anthrax bacillus edema factor (EF), or its combination;
(B) at least a construction, described construction comprises: at least a antigen nucleotide sequence, described antigen nucleotide sequence is expressed at least a antigen protein or antigen protein fragment.
The present invention also can be the biovaccine that a kind of production system of biovaccine from the above mentioned produces.
The present invention comprises that also said biovaccine prevents, alleviates, controls or treat the purposes in following disease, disease or the situation medicine in preparation: flu, fluidity flu, pneumonia, hepatitis, tracheitis, diarrhoea, herpes, endophthalmitis, keratitis, measles, parotitis, German measles and varicella-zoster.
One of this microbial cell also can be Bacillus coli cells, bacillus subtilis cell, or its combination.
Like Fig. 1 or Fig. 2, biovaccine production system of the present invention, this microbial cell 1 can comprise the gene of at least one defective or disappearance.The gene of this defective or disappearance can be in cellular genome 4 or on the plasmid.When this microbial cell is anthrax bacillus cell (Bacillus anthracis) or nontoxic anthrax bacillus cell (Bacillus anthracis Sterne strain); The gene of this defective or disappearance can be anthrax bacillus lethal factor lef gene (LF, lethal factor), anthrax bacillus phosphatidylinositols esterase gene (PI-PLC; Phosphatidylinositol-specific phospholipase C); One of anthrax bacillus edema factor (EF, edma factor), or its combination.Comprise the anthrax bacillus cell or the existing play-by-play (WO 2009/073330, and international filing date is on November 12nd, 2008) in document of nontoxic anthrax bacillus cell of at least one defective or missing gene.
This anthrax bacillus lethal factor gene (LF) can contain the nucleotide sequence of the 331st to 418 of disappearance encoding amino acid sequence.This anthrax bacillus lethal factor gene (LF) can lack the nucleotide sequence of whole this anthrax bacillus lethal factor gene (LF) of coding.
This anthrax bacillus virulence factor gene (PI-PLC) can contain the nucleotide sequence of this anthrax bacillus virulence factor gene (PI-PLC) of whole this anthrax bacillus of disappearance coding.
This anthrax bacillus edema factor (EF) can contain the nucleotide sequence that lacks encoding amino acid sequence the 359th to 601.
Above-mentioned biovaccine production system, this expression vector can be integrated on cellular genome or the plasmid.If use anthrax bacillus cell or nontoxic anthrax bacillus cell, this expression vector can be integrated on this anthrax bacillus cell or this nontoxic anthrax bacillus cellular genome (Fig. 1) or the plasmid (Fig. 2).This expression vector can be integrated in lef (anthrax bacillus lethal factor, the lethal factor) gene on this anthrax bacillus cell or this nontoxic anthrax bacillus cellular genome or the plasmid.
Above-mentioned biovaccine production system, the one or more antigen genes of this antigen nucleotide sequence codified.Any antigen gene that can in microbial cell, express is all applicable to the present invention.This one or more allos of antigen nucleotide sequence codified or isogeneic gene.The one or more heterozygosis antigen genes of this antigen nucleotide sequence codified.The antigen gene of the one or more synthetic of this antigen nucleotide sequence codified.The one or more complete antigen genes of this antigen nucleotide sequence codified, the also fragment of the one or more antigen genes of codified.In a sample, the one or more complete antigen genes of this antigen nucleotide sequence codified.In another sample, the one or more antigen gene fragments of this antigen nucleotide sequence codified.The antigen gene that this is complete or the fragment of antigen gene can be controlled by one or more expression and regulation mechanisms, such as one of promoter, enhancer, inhibition or its combination.
The one or more genes of this antigen nucleotide sequence codified, said gene are selected from down one or more microorganisms of group: adenovirus (Adenovirus), cytomegalovirus (Cytomegalovirus); Dengue virus (Dengue virus), nerpes vinrus hominis's the 4th type (Epstein Barr virus), HAV (Hepatitis A); Herpes simplex virus (Herpes virus), influenza A virus (Influenza A virus), influenza A virus (Influenza A H1N1); Influenza B virus (Influenza B virus), Measles virus (Measles virus), mumps virus (Mumps virus); Parainfluenza virus 1 (Parainfluenza 1), parainfluenza virus 2 (Parainfluenza 2), parainfluenza virus 3 (Parainfluenza 3); Rotavirus (Rotavirus), foot and mouth disease virus, respiratory syncytial virus (Respiratory Syncytialvirus); Rubella virus KOS (Rubella KOS), rubella virus K1S (Rubella K1S), rubella virus K2S (Rubella K2S); Chickenpox one varicella zoster virus (Varicella-Zoster virus), mycoplasma pneumoniae (Mycoplasma pneumoniae), chlamydia trachomatis (Chlamydia trachomatis); One of CPN (Chlamydia pneumoniae), or its combination.
Above-mentioned biovaccine production system, this antigen nucleotide sequence codified part or whole influenza hemagglutinin (HA) gene.
The biovaccine that the present invention also provides a kind of production system of biovaccine from the above mentioned to produce.This biovaccine can be this anthrax bacillus cell (Bacillus anthracis) or this nontoxic anthrax bacillus cell (Bacillus anthracis Sterne strain) and contains any one or a plurality of above-mentioned antigen protein or antigen protein fragment; Also can be the spore 10 of this anthrax bacillus cell or this nontoxic anthrax bacillus cell, said spore contains any one or a plurality of above-mentioned antigen protein or antigen protein fragment (Fig. 1 and Fig. 2).
The biovaccine that biovaccine production system according to the invention produces is used in prevention, controls or treats the purposes in following disease, disease or the situation medicine: flu, fluidity flu, foot and mouth disease (Aphtae epizooticae or foot-and-mouth disease (FMD) or hoof-and-mouth disease), pneumonia, hepatitis, tracheitis, diarrhoea, herpes, endophthalmitis, keratitis, measles, parotitis, German measles and varicella-zoster.
The biovaccine that biovaccine production system according to the invention produces is when being used for such use, and its route of administration is following any: oral, suck, and the Sublingual absorbs, buccal absorption, nasal absorption, urethra absorbs and anal absorbs.Biovaccine according to the invention can mix oral with food.
The biovaccine that biovaccine production system according to the invention produces can be used for the mankind or animal.Animal can comprise poultry and birds or other economic animal, such as chicken, duck, goose, Carnis Coturnicis japonicae, ornamental bird etc.; Domestic animal or other economic animal are such as pig, cattle, horse, sheep, mule etc.; Other mammal is such as monkey, camel, Canis familiaris L., cat, leopard cat etc.
Because microbial cell has the characteristic of quick growth and breeding, biovaccine production system according to the invention has fast high-efficient advantage.
Owing to utilize clone technology, biovaccine production system according to the invention can have extensive applicability, can produce specific biovaccine as required.
Because microbial spore capable of using is as the form of biovaccine, the biovaccine of biovaccine production system according to the invention has the stability of height and permanent preservation effect duration.But long preservation behind the biovaccine microbial spore vacuum drying according to the invention.
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in the restriction scope of the present invention.The experimental technique of unreceipted actual conditions in the following example; Usually according to people such as normal condition such as Sambrook; Molecular cloning: lab guide (New York:Cold Spring Harbor Laboratory Press; 1989) condition described in, or the condition of advising according to manufacturer.Unless otherwise indicated, otherwise percentage ratio and umber calculate by weight.
Only if definition separately, the same meaning that employed all specialties and scientific words and one skilled in the art are familiar with in the literary composition.In addition, any with the institute similar content of putting down in writing or the equalization method and material all can be applicable among the present invention.The usefulness that preferable implementation method described in the literary composition and material only present a demonstration.
Plasmid pSK108 (Fig. 3 A) has whole anthrax bacillus lef gene from the pXO1 plasmid, comprises～nucleotide sequence of 900bp upstream and downstream from the pXO1 plasmid.The cloning vehicle of plasmid pSK108 is pBluescript II.
The restriction endonuclease HincII/SacI fragment of plasmid pSK108 comprises whole anthrax bacillus lef gene nucleic acid sequence of coming from the pXO1 plasmid is cloned into the carrier pUC19 again, constitutes plasmid pSB1 (Fig. 3 B).
Influenza virus A (influenza A/Puerto Rico/8/34) hemagglutinin (HA) (influenzaHA) nucleotide sequence can obtain through PCR from plasmid pT3PR8.This influenza virus A (influenza HA) nucleotide sequence is cloned in the NcoI/BsaAI site of plasmid pSB1, constitutes plasmid plefHa1 (Fig. 3 C).The restriction endonuclease HindIII endonuclease bamhi that contains this influenza virus A (influenza HA) and lef genetic fragment nucleotide sequence of plasmid plefHa1 can be cloned into and be constituted plasmid pSB2 in the HindIII restriction enzyme site of anthrax bacillus plasmid pAT113.Maybe this restriction endonuclease HindIII fragment directly is used to transform anthrax bacillus.The visible following document of this anthrax bacillus plasmid pAT113: Trieu-Cuot P; Carlier C; Poyart-Salmeron C; Courvalin P:An integrative vector exploiting the transposition properties of Tn1545for insertional mutagenesis and cloning of genes from gram-positive bacteria.Gene 106 (1): 21-7,1991.
The derivative strain DMXH of nontoxic anthrax bacillus can be used as F-strain.One section 798bp fragment of the lef gene of this derivative strain DMXH is caused spectinomycin resistance (sper) by the displacement of a spectinomycin (spectinomycin) resistant gene, causes anthrax bacillus lethal factor (lethal factor) lef gene inactivation simultaneously.
Above-mentioned plasmid pSB2 can be used for transforming anthrax bacillus derivative strain DMXH; Described restriction endonuclease HindIII fragment also can directly be used to transform anthrax bacillus derivative strain DMXH.Anthrax bacillus derivative strain DMXH after the conversion is after two reorganization; This spectinomycin (spectinomycin) resistant gene is obtained responsive (spes) derivative strain DMXH 1 of spectinomycin by above-mentioned plasmid pSB2 or the displacement of this restriction endonuclease HindIII fragment resistant gene.Responsive (spes) derivative strain DMXH 1 of this spectinomycin after amplification culture be biological production of vaccine system.
The production of embodiment 3, biovaccine
Responsive (spes) derivative strain DMXH 1 of this spectinomycin can process a large amount of bacillus cultures by a large amount of cultivation of conventional method.
Then; This bacillus culture can place and produce (Nicholson WL under the spore condition of culture; And P.Setlow; Molecular biological methods for B acillus.Sporulation, germination and out-growth ed.I.C.R.H.a.S.M.C. (ed.) .1990, Chichester:John Wiley & Sons Ltd.p.391-450.) formation anthrax bacillus spore.This anthrax bacillus spore drying in vacuum is as the biovaccine of influenza.
The nasal cavity that this anthrax bacillus spore can be inserted the C57BL/6 mice carries out immunity.This anthrax bacillus spore also can be sneaked in the food of C57BL/6 mice mice is carried out immunity.
(avian influenza AI) is caused by A type influenza virus avian influenza A.The HA genetic fragment of A type influenza virus (avian influenza HA) obtains rear clone through PCR and in plasmid pSB1, constitutes plasmid plefHa2.The derivative strain DMXH of nontoxic anthrax bacillus can be used as F-strain.Can cultivate in a large number then and process a large amount of bacillus cultures.Then, this bacillus culture can place and produce spore condition of culture formation anthrax bacillus spore down.This anthrax bacillus spore drying in vacuum is as the biovaccine of influenza.
This anthrax bacillus spore can be sneaked in the food of chicken the chicken crowd is carried out immunity.
All documents in that the present invention mentions are all quoted as a reference in this application, are just quoted such as a reference separately as each piece document.Should be understood that in addition after having read above-mentioned teachings of the present invention, those skilled in the art can do various changes or modification to the present invention, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
Claims (10)
1. biovaccine production system, described system comprises:
(A) at least a microbial cell; Described microbial cell is anthrax bacillus cell (Bacillus anthracis) or nontoxic anthrax bacillus cell (Bacillus anthracis Sterne strain); Its defective or disappearance are selected from following gene: anthrax bacillus lethal factor gene (LF); One of anthrax bacillus virulence factor gene (PI-PLC), anthrax bacillus edema factor (EF), or its combination;
(B) at least a construction, described construction comprises: at least a antigen nucleotide sequence, described antigen nucleotide sequence is expressed at least a antigen protein or antigen protein fragment.
2. biovaccine production system as claimed in claim 1 is characterized in that, (B) in, described construction is an expression vector, this expression vector can exist in described microbial cell or duplicate;
And described antigen nucleotide sequence is integrated in this expression vector and can in described microbial cell, expresses and produces described antigen protein or antigen protein fragment.
3. according to the described biovaccine production system of claim 1, in described anthrax bacillus cell or the nontoxic anthrax bacillus cell:
(a) comprise the nucleotide sequence of anthrax bacillus lethal factor gene (LF), but the 331st to 418 disappearance of the aminoacid sequence of the anthrax bacillus lethal factor of its coding;
(b) do not comprise the nucleotide sequence of anthrax bacillus lethal factor gene (LF)
(c) do not comprise the nucleotide sequence of anthrax bacillus virulence factor gene (PI-PLC);
(d) comprise the nucleotide sequence of anthrax bacillus anthrax bacillus edema factor gene (EF), but the 359th to 601 disappearance of the aminoacid sequence of the anthrax bacillus edema factor of its coding; Or
(e) above (a)-(d) binomial or above combination of binomial arbitrarily.
4. according to the described biovaccine production system of claim 1, described construction can be integrated on described anthrax bacillus cell (Bacillus anthracis) or nontoxic anthrax bacillus cell (Bacillus anthracis Sterne strain) genome or the plasmid.
5. according to the described biovaccine production system of claim 1, described antigen protein or antigen protein fragment derive from one or more microorganisms that are selected from down group: adenovirus (Adenovirus), cytomegalovirus (Cytomegalovirus); Dengue virus (Dengue virus), nerpes vinrus hominis's the 4th type 10 (Epstein Barr virus), HAV (Hepatitis A); Herpes simplex virus (Herpes virus), influenza A virus (Influenza A virus), influenza A virus (Influenza A H1N1); Influenza B virus (Influenza B virus), Measles virus (Measles virus), mumps virus (Mumpsvirus); Parainfluenza virus 1 (Parainfluenza 1), parainfluenza virus 2 (Parainfluenza 2), parainfluenza virus 3 (Parainfluenza 3); Rotavirus (Rotavirus), foot and mouth disease virus, respiratory syncytial virus (Respiratory Syncytialvirus); Rubella virus KOS (Rubella KOS), rubella virus K1S (Rubella K1S), rubella virus K2S (Rubella K2S); Chickenpox one varicella zoster virus (Varicella-Zoster virus), mycoplasma pneumoniae (Mycoplasma pneumoniae), chlamydia trachomatis (Chlamydia trachomatis); CPN (Chlamydia pneumoniae), or its combination.
6. according to the described biovaccine production system of claim 1, described antigen nucleic acid sequence encoding part or total length influenza virus HA (influenza hemagglutinin).
7. a biovaccine is produced by the arbitrary described biovaccine production system of claim 1-6.
8. according to the described biovaccine of claim 7, described biovaccine is:
Described anthrax bacillus cell or nontoxic anthrax bacillus cell, it contains any one or a plurality of above-mentioned antigen protein or antigen protein fragment; Or
The spore of described anthrax bacillus cell or nontoxic anthrax bacillus cell, it contains any one or a plurality of above-mentioned antigen protein or antigen protein fragment.
9. according to the purposes of the described biovaccine of claim 8 in preparing the medicine that prevents, controls or treat following disease, disease or situation: flu, fluidity flu, foot and mouth disease, pneumonia, hepatitis, tracheitis, diarrhoea, herpes, endophthalmitis, keratitis, measles, parotitis, German measles and varicella-zoster.
10. according to the described purposes of claim 9, the route of administration of said medicine is following any: oral, suck, and the Sublingual absorbs, buccal absorption, nasal absorption, urethra absorbs and anal absorbs.
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| CN1324803A (en) * | 2000-05-22 | 2001-12-05 | 首都医科大学 | Efficient production process and application of BPI-Fc recombinant protein |
| CN1641034A (en) * | 2004-01-14 | 2005-07-20 | 中国药品生物制品检定所 | Method for preparing tetanus toxin recombinant antigen and its use |
| KR100690230B1 (en) * | 2005-06-16 | 2007-03-12 | 학교법인 성균관대학 | Vector system for expressing soluble protein with high efficiency |
| WO2008073169A2 (en) * | 2006-08-31 | 2008-06-19 | The Trustees Of The University Of Pennsylvania | Methods, compositions and kits comprising attenuated anthrax vaccines and methods of delivery |
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2010
- 2010-11-15 CN CN2010105454657A patent/CN102462838A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1324803A (en) * | 2000-05-22 | 2001-12-05 | 首都医科大学 | Efficient production process and application of BPI-Fc recombinant protein |
| CN1641034A (en) * | 2004-01-14 | 2005-07-20 | 中国药品生物制品检定所 | Method for preparing tetanus toxin recombinant antigen and its use |
| KR100690230B1 (en) * | 2005-06-16 | 2007-03-12 | 학교법인 성균관대학 | Vector system for expressing soluble protein with high efficiency |
| WO2008073169A2 (en) * | 2006-08-31 | 2008-06-19 | The Trustees Of The University Of Pennsylvania | Methods, compositions and kits comprising attenuated anthrax vaccines and methods of delivery |
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Application publication date: 20120523 |