CN101899416A - Recombinant rat myeloma cell NS0-F10 and application thereof - Google Patents
Recombinant rat myeloma cell NS0-F10 and application thereof Download PDFInfo
- Publication number
- CN101899416A CN101899416A CN2010101847286A CN201010184728A CN101899416A CN 101899416 A CN101899416 A CN 101899416A CN 2010101847286 A CN2010101847286 A CN 2010101847286A CN 201010184728 A CN201010184728 A CN 201010184728A CN 101899416 A CN101899416 A CN 101899416A
- Authority
- CN
- China
- Prior art keywords
- sequence
- monoclonal antibody
- antibody
- derivative
- amino acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Abstract
The invention discloses a recombinant rat myeloma cell NS0-F10 and application thereof. A heavy chain variable region of an antibody has an amino acid residue sequence shown by a sequence 1 in a sequence list, and a light chain variable region of the antibody has an amino acid residue sequence shown by a sequence 3 in the sequence list. Experiments prove that the dissociation constants of a monoclonal antibody secreted by the recombinant rat myeloma cell NS0-F10 and a human tumor necrosis factor alpha (TNFa) are 3nM respectively, and bioactivity EC50 is 0.6nM. The monoclonal antibody and a derivative thereof can specifically recognize a tumor necrosis factor alpha with high affinity, neutralize the biologic activity of the tumor necrosis factor alpha and can be used for treating autoimmune diseases such as rheumatoid arthritis, ankylosing spondylitis and the like.
Description
Technical field
The present invention relates to the monoclonal antibody and the application thereof of anti-tumor necrosis factor.
Background technology
Autoimmune disorder is the chronic disease that threatens human health, because the number that diseases such as rheumatoid arthritis and ankylosing spondylitis disable exceeds traffic accident.
The medicine that is used for the treatment of symptoms of rheumatoid arthritis in early days and keeps patients ' life quality is a non-steroidal anti-inflammatory drugs, and such as acetylsalicylic acid, naproxen and cox 2 inhibitor, these medicines can alleviating pain and inflammatory reaction.Steroid hormone is also united the use reaction that further reduces inflammation with non-steroidal anti-inflammatory drugs, and this combination therapy can obtain stronger short-term antiphlogistic effects, but this effect reduces as time passes and gradually until disappearance.
The medicine of another kind of treatment rheumatoid arthritis is called DMARDS (disease-modifying antirheumatic drugs), and this class medicine is not only relief of symptoms, and can block the advancing of disease process.The most normal medicine that uses is Rheumatrex, and other drug comprises that golden salt, antimalarial drug, Salazosulfamide give a tongue-lashing pyridine (Sulphasalazine), tsiklomitsin and ciclosporin.
Although Rheumatrex is all effective to most of rheumatoid arthritis people, many patients can't tolerate the side effect of this medicine, and about 50% patient finally can stop using this medicine.These patients can use some newer biological response modifier, as leflunomide (Leflunomide, Arava), but its mechanism of action and untoward reaction and Rheumatrex are similar, are used for Rheumatrex as the second line treatment medicine and can't tolerate or treat the patient of failure.
Above-mentioned mention all belong to use old medicine for many years, determined curative effect, usage quantity is big, but owing to be generic drugs, and is cheap, so sales volume is not high.Succeeded in developing a series of immunomodulators in recent years, these immunomodulators belong to patent drugs, the price height, and producer has brought enormous profits for exploitation.The most successful tumour necrosis factor (TNF) inhibitor that surely belongs to can be accepted the tnf inhibitor treatment to the severe rheumatoid arthritis patient in about 30%, and will permeate to moderate patient group toward light.
The present invention is the antigen immune mouse with people TNF recombinant protein, successfully obtains many strains hybridoma, and with the bioactive monoclonal antibody of TNF, wherein a strain avidity reaches 3nM in can secreting, and biological activity EC50 is 0.6nM.This antibody has been enough to the precursor as our antibody drug, and the humanization step is finished at present.
Summary of the invention
An object of the present invention is to provide the monoclonal antibody of secretion specificity in conjunction with human tumor necrosis factor.
Antibody provided by the present invention can be following 1), 2) described antibody:
1) a kind of antibody, its variable region of heavy chain have the amino acid residue sequence shown in the sequence 1 in the sequence table, and its variable region of light chain has the amino acid residue sequence shown in the sequence 3 in the sequence table.
2) monoclonal antibody that obtains by the recombinant rat myeloma cell NS 0-F 10 secretion.
Above-mentioned 1) encoding gene of the encoding gene of the variable region of heavy chain of antibody and variable region of light chain also belongs to protection scope of the present invention described in.
Another object of the present invention provides a kind of clone of secreting specificity in conjunction with the monoclonal antibody of huamn tumor necrosis factory alpha.
The dissociation constant that experimental results show that recombinant rat myeloma cell NS 0-F 10 excretory monoclonal antibody human TNF α is 3nM, and biological activity EC50 is 0.6nM.
Said monoclonal antibody can high-affinity specific recognition tumour necrosis factor, and its biologic activity that neutralizes, and can be used for the treatment of autoimmune disorders such as rheumatoid arthritis and ankylosing spondylitis.
Description of drawings
Fig. 1 detects the dissociation curve (ordinate zou is the absorbance value after the ELISA colour developing) of hybridoma cell strain 3f8 excretory monoclonal antibody and human TNF alpha for ELISA
Fig. 2 is the bioactive mensuration of hybridoma cell strain 3f8.
Embodiment
Following experimental technique if no special instructions, is ordinary method.
Employed reagent etc. if no special instructions, all can be bought from commercial channels among the following embodiment.
MONOCLONAL ANTIBODIES SPECIFIC FOR and the evaluation of embodiment 1, murine hybridoma 3f8 and excretory anti-tumor necrosis factor α (TNF α) thereof
One, the preparation of hybridoma cell strain murine hybridoma 3f8
1, antigenic preparation:
Human TNF alpha and tubercule bacillus (Mycobacterium tuberculosis, MT) fusion rotein of the little peptide of Psts-1 Argine Monohydrochloride 326-344 (DQVHFQPLPPAVVKLSDAL) (acquisition of TNF α-MT): recombinate by PCR method, obtains fusion gene TNF α-MT by the cDNA and the little peptide of the human TNF alpha of will encoding; (Novagen, USA), clone's plasmid increases in DH5 α, again via expressing among the BL21, obtains fusion rotein TNF α to the PET-41a carrier of having deleted the GST coding region with fusion gene cloning.Have 6 * his Tag in the fusion rotein, by the Ni-NTA affinitive layer purification.
2, immunity:
The fusion rotein that obtains with step 1 is as antigen immune NZB/W F1 mouse, immunity is three times altogether: 10 microgram TNF α-MT add the subcutaneous immunity of complete freund's adjuvant (immunity for the first time), and 5 microgram HMGB1-MT add the subcutaneous immunity of incomplete Freund's adjuvant (immunity for the second time) after 14 days.5 microgram HMGB1-MT add the subcutaneous immunity for the third time of incomplete Freund's adjuvant after 14 days; NZB/W F1 mouse after 3 days is got immunocyte in the spleen and rat bone marrow tumour SP2/0 cell with 5: 1 mixed, merges with polyoxyethylene glycol, obtains hybridoma after the HAT screening.
3, hybridoma screening
The preparation of GST-TNF α recombinant protein: the coding region (sequence is shown in sequence in the sequence table 5) of human TNF alpha is cloned into PET-41a carrier (Novagen, USA) EcoR I and HindIII restriction enzyme site, be converted into DH5 α, resistance screening obtains positive colony, extract the plasmid of positive colony, order-checking, the sequence of TNF α and direction of insertion are correct in the plasmid as a result; Change positive plasmid over to e. coli bl21,1mM IPTG abduction delivering; Purifying: have GST Tag in the fusion rotein, by the gsh affinitive layer purification, concrete steps are, with the centrifugal collection of bacterium liquid 5000rpm 10min after inducing, abandon supernatant, with PBS that thalline is resuspended, ultrasonication, the centrifugal 10min of 5000rpm, abandon precipitation, slowly by gsh chromatography column (Sigma), the fusion rotein that will hang on the gsh chromatography column with the gsh elutriant elutes, and obtains GST-TNF α recombinant protein with supernatant.
The secretion specificity detects in conjunction with the hybridoma of TNF Alpha antibodies: as the coating antigen wrapper sheet, hybridoma supernatant (carrying out gradient dilution) is one anti-, HRP link coupled goat anti-mouse igg polyclonal antibody (R﹠amp with GST-TNF α recombinant protein (10 mcg/ml); D Systems) be that the two anti-ELISA that carry out detect.
Subclone: repeatedly clone the hybridoma of secreting specificity antibody with restricted dilution method, obtain the anti-human TNF alpha hybridoma cell strain of hybridoma cell strain murine hybridoma 3f8 at last.
4, MONOCLONAL ANTIBODIES SPECIFIC FOR
(1) preparation method of ascites: Balb/C mouse peritoneal injection pristane 0.5ml/ only after ten days, in mouse peritoneal, treats the hybridoma suspension inoculation to collect ascites after mouse peritoneal has obvious swelling, the centrifuging and taking supernatant.By albumin A/G affinity column (Pierce) antibody purification, 0.1MGlycine/HCl (pH2.5) wash-out
(2) extracorporeal culturing method:
The hybridoma 3f8 that has set up is placed cell culture medium, place 37 ℃ and 5%CO
2Cultivate in the incubator, change cell culture fluid one time, treat that cell concn is greater than 10 every 2d
5Individual/as to stop to change liquid during ml, it is all dead to continue to cultivate cell.1500rpm, centrifugal 10 minutes, collect culture supernatant, supernatant contains high-caliber monoclonal antibody, and-20 ℃ of preservations are standby.Described cell culture medium is for to add foetal calf serum in the DMEM substratum, making the final concentration of foetal calf serum in cell culture medium is 20% (volumn concentration), and the pH of described cell culture medium is 7.4.
Purifying: the supernatant that vitro culture is obtained carries out following purifying, above-mentioned supernatant is slowly passed through albumin A/G affinity column (Pierce), the target protein of using 0.1M Glycine/HCl (pH2.5) will hang on the chromatography column again elutes, and obtains the albumen behind the purifying.
5, the Performance Detection of hybridoma 3f8 excretory antibody
(1) ELISA detects the dissociation curve of hybridoma cell strain 3f8 excretory monoclonal antibody and HMGB1.
Experimental technique: (preparation method sees that step 1) (2ug/ml) is as envelope antigen with TNF α-MT, anti-(with 160ug/ml is initial concentration to the hybridoma 3f8 excretory monoclonal antibody of step 4 preparation purifying as one, two times two times are diluted), goat anti-mouse igg-HRP (R﹠amp; D) (1: 2000) is that the two anti-ELISA that carry out measure.
3 repetitions are established in experiment, the result as shown in Figure 1,50% corresponding antibody concentration of ELISA full-scale reading is the dissociation constant of antibody.The dissociation constant that shows hybridoma cell strain 3f8 excretory monoclonal antibody and TNF α is 3nM.The X-coordinate of Fig. 1 is the concentration of hybridoma cell strain 3f8 excretory monoclonal antibody.
(2) antibody subtype identification experiment: with GST-TNF α recombinant protein (10 mcg/ml) wrapper sheet, the hybridoma supernatant is one anti-, HRP link coupled rat anti-mouse IgG1, IgG2a, or IgG2b monoclonal antibody (BD Pharmingen) is that the two anti-ELISA that carry out detect.Experimental results show that hybridoma cell strain 3f8 excretory monoclonal antibody is the IgG2b hypotype.
(3) the bioactive mensuration of antibody:
3 repetitions are established in experiment, the result as shown in Figure 2, biological activity EC50 is 0.6nM.
The preparation of embodiment 2, anti-TNF alpha humanized antibody pCI-DHFRr-gpt-hu3f8
Murine antibody can produce immunological rejection in human body, therefore can only be used for the treatment of acute disease, in case produce the anti-antibody at murine antibody in the human body, will lose efficacy as the murine antibody of medicine.In order to overcome this shortcoming, the present invention has carried out humanization to murine antibody, has prepared humanized antibody ch-TNF α.The variable region sequences of this antibody is from murine antibody-hybridoma cell strain 3f8 excretory monoclonal antibody, and the constant region sequence is from the human IgG1.This antibody has kept the antigen-binding specificity of mouse monoclonal antibody, has reduced inductive immunological rejection in human body simultaneously.
The step of preparation humanized antibody is as follows:
Separation and purification mRNA from hybridoma cell strain 3f8 is with the synthetic first chain cDNA of oligo-dT.With PCR method increase respectively light chain of antibody and variable region of heavy chain, the light chain primer is 5 ' GAY ATT GTGMTS ACM CAR WCT MCA 3 ' and 5 ' CTC CAG ATG TTA ACT GCT CAC 3 '; The heavy chain primer is: 5 ' ATG SAR GTN MAGCTG SAG SAG TC 3 ' and 5 ' GGT CAA GGTCAC TGG CTC AGG3 '.Wherein, R=G or A, Y=T or C, M=A or C, S=G or C, W=T or A, N=G or A or C or T.The PCR product is cloned into respectively in the T carrier checks order.The result shows the nucleotide sequence that the encoding gene of the variable region of heavy chain of hybridoma cell strain 3f8 excretory monoclonal antibody has sequence 6 in the sequence table, and coding has the variable region of heavy chain of the amino acid residue sequence of sequence 7 in the sequence table; The nucleotide sequence that the encoding gene of the variable region of light chain of hybridoma cell strain 3f8 excretory monoclonal antibody has sequence 8 in the sequence table, coding has the variable region of light chain of the amino acid residue sequence of sequence 9 in the sequence table.
With the antibody heavy chain variable region gene and the chain variable region gene humanization that obtain, the encoding gene of antibody heavy chain variable region has the nucleotide sequence of sequence 2 in the sequence table, the amino acid residue sequence that coding has sequence 1 in the sequence table.The encoding gene of antibody chain variable region has the nucleotide sequence of sequence 4 in the sequence table, the amino acid residue sequence that coding has sequence 3 in the sequence table.
The chain variable region gene of humanized monoclonal antibody 3f8 is connected with light chain carrier pCI-gpt's: with PCR method the humanization variable region of light chain is introduced Sal I and two restriction enzyme sites of Xba I, primer is 5 ' ccc agg gtc gac cgg aga cat tgt gct cac cca gtc tcc agcttc 3 ' and 5 ' gaa ttt cta gaa gac aat agt gaa aaa tta ctt tgc agc atc3 '.The chain variable region gene of humanized monoclonal antibody 3f8 can be inserted among the carrier pCI-gpt by Sal I and two restriction enzyme sites of Xba I, obtains pCI-gpt-hu3f8-VL
The heavy chain variable region gene of humanized monoclonal antibody 3f8 is connected with light chain carrier pCI-DHFRr's: with PCR method the humanization variable region of light chain is introduced Not I and two restriction enzyme sites of Xba I, primer is 5 ' ggt ggc ggc cgc aac agg tgc cca ctc cca ggt cca act ggtcca g 3 ' and 5 ' aag ctc tag aag aca ata gtg aaa aat tac tta ccg cttgag gag ac3 '.The heavy chain chain variable region gene of humanized monoclonal antibody 3f8 can be inserted among the carrier pCI-DHFRr by SalI and two restriction enzyme sites of Xba I, obtains pCI-DHFRr-hu3f8-VH.
Variable region part among the pCI-DHFRr-hu3f8-VH and constant region are partly downcut with restriction endonuclease NotI/Xho I, be inserted in two restriction enzyme sites of Not I/Xho I of pCI-gpt-hu3f8-VL, thereby construct humanized antibody pCI-DHFRr-gpt-hu3f8.
To contain humanized antibody expression carrier pCI-DHFRr-gpt-hu3f8 with the method for electrotransfection imports among the mammalian cell NS/0 (ECACC purchase).(Mycophenolate) screens transformant in the substratum that contains xanthine (Xanthine) with mycophenlate mofetil, obtains the cell strain of stable transfection, and note is made NS/3f8.
The cultural method of cell NS/3f8: with RPMI 1640 culture medium culturing that contain 10% foetal calf serum.
Sequence table
Qvqlvqsgselkkpgasvkisckasgytftnygmnwvrqapgkglewmgwintytgeptyadd
fkgrfvfsldtsvstaylqisslkaedtavyycarrrsydydvamdywgqgtlvtvss
Sequence 2
Caggtccaactggtccagtctggatctgagctgaagaagcctggagcttctgtcaagatctcc
tgcaaggcttctgggtataccttcacaaactatggaatgaactgggtgcggcaggctccagga
aagggtttagagtggatgggctggataaacacctacactggagagccaacatatgctgatgac
ttcaagggacggtttgtcttctctttggatacctctgtcagcactgcctatttgcagatctcg
tcgctcaaagctgaggacacagctgtctattactgtgcaagaagaagaagctatgattacgac
gtggctatggactactggggtcaaggaaccctagtcaccgtctcctca
Divmtqspdslavslgqratincrasesvdsygnyfmhwyqqkpgqppklliyrasnlesgvp
drfsgsgsgtdftltisslqaedvatyycqqsneepltfgqgtkleikradaa
Sequence 4
Gacattgtgatgacccagtctccagattctttggctgtgtctctagggcagagggccaccata
aactgcagagccagtgaaagtgttgatagttatggcaattattttatgcactggtatcagcag
aaaccaggacagccacccaaactcctcatctatcgtgcatccaacctagaatctggggtccct
gacaggttcagtggcagtgggtctgggacagacttcaccctcaccattagtagtctgcaggct
gaagatgttgcaacctattactgtcaacaaagtaatgaggagcctctcacgttcggccagggg
acaaagttggaaataaaacgggctgatgctgca
ctccctcagcaaggacagcagaggaccagctaagagggagagaagcaactacagaccccccct
gaaaacaaccctcagacgccacatcccctgacaagctgccaggcaggttctcttcctctcaca
tactgacccacggctccaccctctctcccctggaaaggacaccatgagcactgaaagcatgat
ccgggacgtggagctggccgaggaggcgctccccaagaagacaggggggccccagggctccag
gcggtgcttgttcctcagcctcttctccttcctgatcgtggcaggcgccaccacgctcttctg
cctgctgcactttggagtgatcggcccccagagggaagagttccccagggacctctctctaat
cagccctctggcccaggcagtcagatcatcttctcgaaccccgagtgacaagcctgtagccca
tgttgtagcaaaccctcaagctgaggggcagctccagtggctgaaccgccgggccaatgccct
cctggccaatggcgtggagctgagagataaccagctggtggtgccatcagagggcctgtacct
catctactcccaggtcctcttcaagggccaaggctgcccctccacccatgtgctcctcaccca
caccatcagccgcatcgccgtctcctaccagaccaaggtcaacctcctctctgccatcaagag
cccctgccagagggagaccccagagggggctgaggccaagccctggtatgagcccatctatct
gggaggggtcttccagctggagaagggtgaccgactcagcgctgagatcaatcggcccgacta
tctcgactttgccgagtctgggcaggtctactttgggatcattgccctgtgaggaggacgaac
atccaaccttcccaaacgcctcccctgccccaatccctttattaccccctccttcagacaccc
tcaacctcttctggctcaaaaagagaattgggggcttagggtcggaacccaagcttagaactt
taagcaacaagaccaccacttcgaaacctgggattcaggaatgtgtggcctgcacagtgaagt
gctggcaaccactaagaattcaaactggggcctccagaactcactggggcctacagctttgat
ccctgacatctggaatctggagaccagggagcctttggttctggccagaatgctgcaggactt
gagaagacctcacctagaaattgacacaagtggaccttaggccttcctctctccagatgtttc
cagacttccttgagacacggagcccagccctccccatggagccagctccctctatttatgttt
gcacttgtgattatttattatttatttattatttatttatttacagatgaatgtatttatttg
ggagaccggggtatcctgggggacccaatgtaggagctgccttggctcagacatgttttccgt
gaaaacggagctgaacaataggctgttcccatgtagccccctggcctctgtgccttcttttga
ttatgttttttaaaatatttatctgattaagttgtctaaacaatgctgatttggtgaccaact
gtcactcattgctgagcctctgctccccaggggagttgtgtctgtaatcgccctactattcag
tggcgagaaataaagtttgcttagaaaagaa
Sequence 6
Cagtctggacctgagctgaagaagcctggagagacagtcaagatctcctgcaaggcttctggg
tataccttcacaaactatggaatgaactgggtgaagcaggctccaggaaagggtttaaagtgg
atgggctggataaacacctacactggagagccaacatatgctgatgacttcaagggacggttt
gccttctctttggaaacctctgccagcactgcctatttgcagatcaacaacctcaaaaatgag
gactcggctacatatttctgtgcaggaagaagaagctatgattacgacgtggctatggactac
tggggtcaaggaacctcagtcaccatctcctca
qsgpelkkpgetvkisckasgytftnygmnwvkqapgkglkwmgwintytgeptyaddfkgrf
afsletsastaylqinnlknedsatyfcagrrsydydvamdywgqgtsvtiss
Sequence 8
gacattgtgctcacccagtctccagcttctttggctgtgtctctagggcagagggccaccata
tcctgcagagccagtgaaagtgttgatagttatggcaattattttatgcactggtatcagcag
aaaccaggacagccacccaaactcctcatctatcgtgcatccaacctagaatctgggatccct
gccaggttcagtggcagtgggtctgggacagacttcaccctcaccattaatcctgtggaggct
gatgatgttgcaacctattactgtcaacaaagtaatgaggagcctctcacgttcggctcgggg
acaaagttggaaataaaacgggctgatgctgca
divltqspaslavslgqratiscrasesvdsygnyfmhwyqqkpgqppklliyrasnlesgip
arfsgsgsgtdftltinpveaddvatyycqqsneepltfgsgtkleikradaa
Claims (10)
1. recombinant rat myeloma cell NSO-F10 CGMCC No.3846.
2. recombinant rat myeloma cell NSO-F10 CGMCC No.3846 excretory monoclonal antibody and derivative thereof, the derivative of described monoclonal antibody is single-chain antibody, Fab fragment or people-mouse chimeric antibody.
3. monoclonal antibody according to claim 2 and derivative thereof, it is characterized in that: described recombinant rat myeloma cell NSO-F10 CGMCC No.3846 excretory monoclonal antibody, by the amino acid residue sequence that the variable region of heavy chain of described monoclonal antibody deutero-Fab fragment or people-mouse chimeric antibody has sequence 1 in the sequence table, variable region of light chain has the amino acid residue sequence of sequence 2 in the sequence table.
4. monoclonal antibody derivative according to claim 3, it is characterized in that: by the amino acid residue sequence that the light chain of described monoclonal antibody deutero-people-mouse chimeric antibody has sequence 8 in the sequence table, heavy chain has the amino acid residue sequence of sequence 9 in the sequence table.
5. monoclonal antibody according to claim 2 and derivative thereof is characterized in that: the amino acid residue sequence that has sequence 3 in the sequence table by described monoclonal antibody deutero-single-chain antibody.
6. the encoding gene of each described monoclonal antibody and derivative thereof among the claim 2-5.
7. encoding gene according to claim 6, it is characterized in that: by the nucleotide sequence that the light chain encoding gene of described monoclonal antibody deutero-people-mouse chimeric antibody has sequence 6 in the sequence table, the heavy chain encoding gene has the nucleotide sequence of sequence 7 in the sequence table.
8. each described monoclonal antibody and derivative thereof the application in the preparation medicine among the claim 2-5, described medicine is for treating by the medicine of the inflammatory reaction of people TNF-alpha participation, by the autoimmune disorder of people TNF-alpha participation or the medicine of organ damage disease.
9. application according to claim 8 is characterized in that: described inflammatory reaction is a septicemia, systemic inflammatory response syndrome or acute lung injury; Described autoimmune disorder is a rheumatoid arthritis, diabetes or lupus erythematosus; Described organ damage disease is a myocardial infarction, cerebral infarction, medicine or viral liver injury.
10. each described monoclonal antibody and the application of derivative in surveyor TNF-alpha thereof among the claim 2-5.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010101847286A CN101899416A (en) | 2010-05-27 | 2010-05-27 | Recombinant rat myeloma cell NS0-F10 and application thereof |
| CN201110140392.8A CN102260648B (en) | 2010-05-27 | 2011-05-27 | Recombinant rat myeloma cell NS 0-F 10 and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010101847286A CN101899416A (en) | 2010-05-27 | 2010-05-27 | Recombinant rat myeloma cell NS0-F10 and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101899416A true CN101899416A (en) | 2010-12-01 |
Family
ID=43225338
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010101847286A Pending CN101899416A (en) | 2010-05-27 | 2010-05-27 | Recombinant rat myeloma cell NS0-F10 and application thereof |
| CN201110140392.8A Expired - Fee Related CN102260648B (en) | 2010-05-27 | 2011-05-27 | Recombinant rat myeloma cell NS 0-F 10 and application thereof |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110140392.8A Expired - Fee Related CN102260648B (en) | 2010-05-27 | 2011-05-27 | Recombinant rat myeloma cell NS 0-F 10 and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN101899416A (en) |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1388135A (en) * | 2001-11-07 | 2003-01-01 | 林琳 | Monoclonal antibody capable of priming cancer cell death |
| CN100427505C (en) * | 2004-08-19 | 2008-10-22 | 中国医学科学院基础医学研究所 | Receptor DR5 monoclonal antibody (AD5-10) resisting human tumor necrosin related apoptosis inducing ligand and its prepn and use |
| CN101560500B (en) * | 2008-04-17 | 2010-12-15 | 中国人民解放军军事医学科学院基础医学研究所 | Cell strain LaDR5 for secreting tumor necrosis factor receptor induced antibody |
| CN101684156B (en) * | 2008-09-27 | 2011-12-28 | 苏州工业园区晨健抗体组药物开发有限公司 | Human TNF alpha monoclonal antibody, PEGylation nanoparticle and application thereof |
-
2010
- 2010-05-27 CN CN2010101847286A patent/CN101899416A/en active Pending
-
2011
- 2011-05-27 CN CN201110140392.8A patent/CN102260648B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN102260648A (en) | 2011-11-30 |
| CN102260648B (en) | 2016-06-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111051347B (en) | TIGIT antibody, antigen-binding fragment thereof, and medical use thereof | |
| EP3753951A1 (en) | Anti-b7-h4 antibody, antigen-binding fragment thereof and pharmaceutical use thereof | |
| CN110914304A (en) | CD96 antibody, antigen binding fragment thereof and medical application | |
| EP3819313A1 (en) | Bispecific antibody and use thereof | |
| TWI635097B (en) | Novel anti-human tslp receptor antibody | |
| CN112041347A (en) | Antibodies that bind human IL-4R, methods of making, and uses thereof | |
| CN113993903A (en) | Tetravalent bispecific antibody for resisting PD-1 and PD-L1 | |
| CN114591434B (en) | anti-Siglec 15 antibody and preparation method and application thereof | |
| ES2903412T3 (en) | Anti-IL-22R antibodies | |
| CN111615519A (en) | Monoclonal antibody binding to human IL-5, preparation method and application thereof | |
| CN112513088B (en) | anti-OX 40 antibodies, antigen binding fragments thereof, and medical uses thereof | |
| CN110461874B (en) | anti-GITR antibodies, antigen binding fragments thereof, and medical uses thereof | |
| EP3683234A1 (en) | Il-6r antibody and antigen binding fragment thereof and medical use | |
| CN115298216A (en) | Antibody or antigen binding fragment thereof, preparation method and medical application thereof | |
| JP7256266B2 (en) | ANTIBODY BINDING HUMAN IL-1β, PREPARATION METHOD AND USE THEREOF | |
| WO2015113494A1 (en) | Bifunctional fusion protein, preparation method therefor, and use thereof | |
| EP1972640A1 (en) | Apoptosis inducing antibodies | |
| CN101899416A (en) | Recombinant rat myeloma cell NS0-F10 and application thereof | |
| WO2021218574A1 (en) | Antibody binding to human ngf, and preparation method therefor and use thereof | |
| CN114380906B (en) | anti-IL-17A single-domain antibody and application thereof | |
| WO2021241616A1 (en) | Anti-human nr1 antibody derivative | |
| CA3221903A1 (en) | Anti-upar antibodies and uses thereof | |
| CN114591432A (en) | anti-TNF alpha single domain antibodies and uses thereof | |
| KR20220022226A (en) | Il-33 binding antibodies or antigen binding fragments thereof | |
| CN116063480A (en) | Anti-cytokeratin 18 antibody, preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Open date: 20101201 |