CN106866823A - A kind of nano antibody of anti-Her2 - Google Patents

A kind of nano antibody of anti-Her2 Download PDF

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CN106866823A
CN106866823A CN201710136524.7A CN201710136524A CN106866823A CN 106866823 A CN106866823 A CN 106866823A CN 201710136524 A CN201710136524 A CN 201710136524A CN 106866823 A CN106866823 A CN 106866823A
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李洪利
凌志
庄文超
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Health (beijing) Biotechnology Co Ltd
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    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/569—Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
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    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
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Abstract

The invention discloses a kind of nano antibody of anti-Her2, the amino acid coding of the antibody：SEQ ID NO：8 arrive SEQ ID NO：14, encode the DNA sequence dna of above-mentioned amino acid sequence：SEQ ID NO：1 to SEQ ID NO：7.

Description

A kind of nano antibody of anti-Her2
Technical field
The invention belongs to biomedical or biological pharmacy technical field, it is related to a kind of nano antibody for being directed to Her2.
Background technology
Human epidermal growth factor receptor-2 (Her2) is to study one of more thorough gene in breast cancer so far, in The eighties in 20th century is independently found by three research groups respectively.The overexpression of Her2 genes not only develops phase with the generation of tumour And be a weight of neoplasm targeted therapy medicament selection outside the Pass, or an important clinical treatment monitoring and prognostic indicator, Want target spot.Serum Her2 has with histology Her2, the tumor load of patient with breast cancer, lymph node status and necessarily associates, and can Certain influence can be produced on chemotherapy or endocrine therapy curative effect, it may be possible to an independent Prognostic Factors.
Human epidermal growth factor acceptor (human epidermal growth factor receptor) family has 4 Member：EGFR, Her2 (ErbB-2), ErbB3 and ErbB4, wide expression is in epithelium, interstitial and nerve Tissue, they participate in the cellular signal transduction pathways of activation a series of complex under physiological status, regulate and control the life of normal tissue cell The important physiology courses such as long, division, differentiation.However, studies have shown that the generation of some tumours, development and malignant behaviors with The unconventionality expression and activation (Albanell J, CodonyJ, Rovira A, Mellado B, Gasc ó n in close relations of EGFR families P.Mechanism of action of anti-Her2monoclonal antibodies:scientific update on trastuzumab and2C4.Adv Exp Med Biol.2003；532:253-68).Her2 does not have native ligand, in monomer It is inactive under state, tyrosine kinase activation can be made when dimer is formed, MAPK and PI3K/Akt paths are activated, ultimately result in Tumor cell proliferation and survival (Hudis CA.Trastuzumab-mechanism of action and use in clinical practice.N Engl J Med.2007Jul5；357(1):39-51).
Belgian scientist reports in Nature first within 1993：Antibody in camel blood, has half not have light chain, And more making people pleasantly surprised, " heavy chain antibody " of these missing light chains can closely be tied as normal antibody with the target such as antigen Close, the mutual adhesion unlike scFv (artificial reconstructed single chain antibody fragments) in addition, or even assemble blocking.This antibody is only wrapped CH2 and CH3 area containing a weight chain variable district and two routines, it is often more important that the VHH areas (weight individually cloned and express Chain variable region) there is good structural stability and antigen-binding activity, molecular weight is the 1/10 of common antibody, so VHH Also referred to as Nanobody (nano antibody)；At the same time nano antibody chemical property is also more flexible, good stability, soluble high, Expression is easy and is readily available, and is easily coupled other molecules, therefore have using nano antibody technical research Her2 detection reagents Wide prospect.
The content of the invention
Goal of the invention：The technical problems to be solved by the invention are to provide a kind of nano antibody for Her2 epitopes, together When provide the nano antibody coding amino acid sequence and nucleotide sequence.
Technical scheme：To achieve the above object, the first aspect of the present invention, there is provided a kind of nano antibody of Her2 VHH chains, the amino acid sequence of the FR being selected from the group：SEQ ID NO：VHH1 shown in 1, SEQ ID NO：VHH2 shown in 2, SEQ ID NO：VHH3 shown in 3, SEQ ID NO：VHH4 shown in 4；Or SEQ ID NO：VHH5 shown in 5, SEQ ID NO：VHH6 shown in 6, SEQ ID NO：VHH7 shown in 7.
A kind of aspect of the present invention the 2nd, there is provided DNA molecular, it encodes the protein being selected from the group：It is of the present invention The VHH chains of the nano antibody of Her2, or Her2 nano antibodies of the present invention.Preferably, described DNA molecular, its feature It is that it has the DNA sequence dna being selected from the group：SEQ ID NO：8 to SEQ ID NO：14
3rd aspect of the invention, there is provided a kind of expression vector, its NO of ID containing SEQ：8 to SEQ ID NO：Shown in 14 Nucleotide sequence.
4th aspect of the invention, there is provided Her2 nano antibodies of the present invention are used to detect the purposes of Her2.
Beneficial effect：Compared with prior art, advantages of the present invention is as follows：The present invention by Her2 antigen immune alpacas, with Fluidic cell screening is carried out using the camel PBLC afterwards, obtains extracting RNA after positive findings, carry out reverse transcription Obtain cDNA.So as to obtain for the specific nano antibody genes of Her2, during this gene gone into Escherichia coli, so as to build Having stood can be in the strain of the nano antibody of E. coli.
Brief description of the drawings
Fig. 1 is the gene electrophoretogram of nano antibody；Wherein swimming lane 1 is DL2000DNA molecular criterias, DL2000 (2000bp, 1000bp, 750bp, 500bp, 250bp, 100bp) swimming lane 2-8 be PCR amplification antibody heavy chain variable region fragment
Fig. 2 is the Her2 nano antibodies of expression, through the electrophoresis of the SDS-PAGE after nickel post resin gel affinitive layer purification Figure；Wherein swimming lane 5 is protein molecular standard, and swimming lane 1-4 and 6-8 are broken the total crude extract sample of albumen after bacterium.Protein molecular Standard is：
116.0KD, 66.2KD, 45.0KD, 35.0KD, 25.0KD, 18.4KD, 14.4KD. destination protein are about 20KD.
Specific embodiment
Her2 antigen immune alpacas are carried out fluidic cell sieve by the present invention followed by the camel PBLC Choosing, obtains extracting RNA after positive findings, carries out reverse transcription and obtains cDNA.So as to obtain for the specific nanometers of Her2 Antibody gene, during this gene gone into Escherichia coli, can be in the strain of the nano antibody of E. coli so as to establish.
With reference to specific embodiment, the present invention is expanded on further.
Embodiment 1：It is directed to the structure in the nano antibody library of Her2：
(1) by purchased from the Her2 antigens of ACROBiosystems companies, its sequence is GenBank:Aaa75493, concentration is 100 micrograms per millilitres, it is immune every time to mix 100 microgram Her2 in equal volume with Freund's adjuvant, an alpaca is immunized, on every Mondays It is secondary, it is immunized 4 times altogether, except first time uses complete Freund's adjuvant, residue all uses not formula incomplete adjuvant, immunologic process several times Moderate stimulation B cell expresses the nano antibody of antigentic specificity.
After (2) 4 immune end, separating periphery blood monocytic cell, airflow classification positive cell simultaneously extracts total serum IgE, reference The RNA extracts kits that QIAGEN companies provide.(3) tried according to Super-Script III FIRST STRANDSUPERMIX Agent box specification, the RNA reverse transcriptions that will be extracted expand VHH chains into cDNA and using sleeve type PCR,
First round PCR：
Sense primer：CAGGTGAAGGTCATCGARTC
Downstream:GATGCTCTTGTGACTCAGGAATC
Amplification obtains single-stranded and double-strand the weight chain variable district-constant region (VH-CH) of alpaca, 53 DEG C of annealing, 35 circulations；
Second wheel PCR：
Template is made with first round PCR primer,
Sense primer：
GGAATTCCATATGGATTATAAAGATGATGATAAACGCAGAGACAGTGACCAGAGT
Anti-sense primer：CCACGATTCTGCGGCCGCTTAATCCAGCTGACTCAGCC
Amplification obtains the single-stranded VHH of alpaca, and purpose fragment is reclaimed in 53 DEG C of annealing, 35 circulations, as a result as Fig. 1 shows.From a left side It is respectively to right DNA bands：First is purpose gene for molecule Marker, 2-8 the swimming band of DL2000.
Embodiment 2：Nano antibody is in Host Strains expression in escherichia coli, purifying：
1. construction of expression vector
The nano antibody DNA fragmentation double digestion of synthesis is connected into expression vector pET28a, in expression place after sequencing is correct Expressed in master.The digestion condition of expression vector is carrier (1ug) 35 μ L, 10 times of fast μ L of enzyme cutting buffer solution 10 of concentration, two kinds of enzymes Each 2 μ L of NdeI, NotI, water 51 μ L, 37 DEG C of water-baths 20min, 80 DEG C of inactivation 10min, agarose gel electrophoresis reclaims quantitative.DNA Fragment digestion condition be DNA fragmentation 1ug 20 μ L, 10 times of fast enzyme cutting buffer solution 10 μ L, two kinds of each 2 μ L of enzyme NdeI, NotI of concentration, Water 66 μ L, 37 DEG C of water-baths 20min, 80 DEG C of inactivation 10min, agarose gel electrophoresis reclaims quantitative.Expression vector connects with DNA fragmentation Narrow bars part：Expression vector (50ng/ μ L) 1 μ L, the μ L of DNA fragmentation (40ng/ μ L) 1.5,10 times of T4 DNAs of concentration delay The μ L of 1 μ L, T4 deoxyribonucleic acid ligase of fliud flushing 1, water 5.5 μ L, room temperature 1h connection, are transferred to BL21 E. coli competents.
2. induced expression
The correct sample of sequence is carried out into induced expression, expression induction step is：Correct sequence samples bacterium solution and 2YT are trained Support base and press 1:100 dilution, 37 DEG C, 180~200rpm carries out shaking bacterium, shake to bacterium solution OD600 values be 0.6~1.0 when, take out on a small quantity Bacterium solution, adding the IPTG of final concentration of 1mmol/L carries out 3~4h of induced expression, collects bacterium solution, empty carrier pet28a bacterium solutions according to The identical expression induction step operation of above-mentioned correct sample bacterium solution；With the empty carrier pet28a bacterium solutions, the empty carrier of induction that do not induce Pet28a bacterium solutions and the correct sample bacterium solution not induced carry out polyacrylamide gel electrophoresis experiment as control, as a result show IPTG can induce expression of the AntiCD3 McAb e nano antibodies in pet28a bacterium solutions.
3. purifying protein
By IPTG induction, expression after bacterium solution in centrifuge 7000rpm, be centrifuged 10min collects thallines, use buffer solution PBS (PH7.4) is cleaned once, and thalline is collected by centrifugation again；Bacterium solution is blown and beaten with the PBS (PH7.4) of 10mL is mixed, be placed in ice On, it is put into Ultrasonic cell smash, shake time 5s, off time 5s are sent out by ultrasound, protect temperature 60 C, ultrasonic power 10% program ultrasonication 10min, the bacterium solution after crushing is put in 7000rpm in centrifuge, 10min is centrifuged, by supernatant Affinitive layer purification is carried out, is saved backup.
The Her2 antibody ELISAs immunocompetence of embodiment 3 is detected
Her2 antigen coating buffers PH (9.6) is diluted to 2ng/ μ L, is added in ELISA Plate by the μ L of every hole 100, is positioned over 4 DEG C refrigerator overnight, carries out ELISA experiments for second day, and ELISA steps are as follows：
Step 1, by the μ L of every hole 200 PBS-T add coating after ELISA Plate cleaned, clean 5 times, each 3min, Cleaning fluid is got rid of, ensures that washing lotion is all blotted in clappers on paper handkerchief；
Step 2, by the μ L of every hole 200 in ELISA Plate add 5% skimmed milk power, room temperature closing 1h；
Step 3, cleaned by cleaning step in step 1, then, successively by the blank liquid of 100 μ L, negative right It is added in ELISA Plate according to, Her2 antibody after purification, is incubated at room temperature 2h；
Step 4, cleaned by cleaning step in step 1, then, added what HRP was marked to ELISA Plate by the μ L of every hole 100 Tag antibody, is incubated at room temperature 1h；
Step 5 is cleaned by cleaning step in step 1, and the TMB of 100 μ L, avoid light place are then added to every hole 10min；
Step 6, added to every hole 50 μ L 2mol/L sulfuric acid solution, be put in ELISA Plate in ELIASA by terminating reaction, The reading under the conditions of 450nm.
Result is as follows：
Wherein, negative control is：The empty carrier nutrient solution of not connected antibody gene
Blank is：PBS washing lotions
MCF-7 SK-br-3 cells are purchased from National Cell resource sharing platform
Finally it should be noted that：Obviously, above-described embodiment is only intended to clearly illustrate the application example, is excellent The embodiment of choosing.And not to the restriction of implementation method.For those of ordinary skill in the field, in described above On the basis of can also make other changes in different forms.There is no need and unable to give thoroughly all of implementation method Lift.And the obvious change thus amplified out or among changing still in the protection domain of the application type.
SEQUENCE LISTING
<110>Kang Zhong（Beijing）Bio tech ltd
<120>A kind of nano antibody of anti-HER2
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<170> PatentIn version 3.3
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atgggtcagg tgcagctggt ggagtctggg ggaggcttgg tgcaggctgg ggactctctg 60
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caggctatag ggaaggagcg tgaatttgta ggcgctgtta gctggacgag cacagaaaca 180
tcctatgcgg acgctgtgaa ggggcgattc accatctcca gagacaacgc caagaacacg 240
gtgtatctgc aaatgaacag cctgaagcct gaggacacgg ccgtttatac ttgtgcagca 300
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gtgtatctgc aaatgaacag cctgaagcct gaggacacgg ccgtttatac ttgtgcagca 300
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caggctccag ggaaggagcg tgagtttgta gcagctatta gctggtgtgg tggtgtcaca 180
ttctatgcaa actccgtgaa gggccgattc accatctcca gagacaacgc caagaacacg 240
gtgtatctgc aaatgaacag cctgaaacca gaggacacgg ccgtctatta ctgtaatgtg 300
aagggcctta cgaacgtagc tggaacgtgg gtttcgggtg actactgggg ccaggggacc 360
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caggctgtag ggaagcagcg cgagttggtc gcggttgtta ctaggagtgc tggcgcccgc 180
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tatctgcaaa tgaacagcct gaaacctgag gacacggccg tttattactg tgcactgact 300
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gtatatctgc aaatgaacag cctgaaacct gaggacacgg ccgtttatta ctgtgcagct 300
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gtgtatctgc aaatgaacag cctgaagcct gaggacacgg ccgtttatac ttgtgcagca 300
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Met Gly Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala
1 5 10 15
Gly Asp Ser Leu Arg Leu Ser Ala Ala Ser Gly Pro Ser Phe Ser Asp
20 25 30
Tyr Ala Val Gly Trp Phe Arg Gln Ala Ile Gly Lys Glu Arg Glu Phe
35 40 45
Val Gly Ala Val Ser Trp Thr Ser Thr Glu Thr Ser Tyr Ala Asp Ala
50 55 60
Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val
65 70 75 80
Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Thr
85 90 95
Cys Ala Ala Thr Arg Thr Tyr Gly Phe Thr Ser Arg Tyr Gln Thr Asn
100 105 110
Tyr Glu Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser Glu Pro
115 120 125
Lys Thr Pro Lys Pro Gln
130
<210> 9
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<212> PRT
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Met Gly Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala
1 5 10 15
Gly Asp Ser Leu Arg Leu Ser Ala Ala Ser Gly Pro Ser Phe Ser Asp
20 25 30
Tyr Ala Val Gly Trp Phe Arg Gln Ala Ile Gly Lys Glu Arg Glu Phe
35 40 45
Val Gly Ala Val Ser Trp Thr Ser Thr Glu Thr Ser Tyr Ala Asp Ala
50 55 60
Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val
65 70 75 80
Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Thr
85 90 95
Cys Ala Ala Thr Arg Thr Tyr Gly Phe Thr Ser Arg Tyr Gln Thr Asn
100 105 110
Tyr Glu Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser Glu Pro
115 120 125
Lys Thr Pro Lys Pro Gln
130
<210> 10
<211> 134
<212> PRT
<213> Vicugna pacos
<400> 10
Met Gly Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro
1 5 10 15
Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser
20 25 30
Ser Tyr Arg Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu
35 40 45
Phe Val Ala Ala Ile Ser Trp Cys Gly Gly Val Thr Phe Tyr Ala Asn
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr
65 70 75 80
Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Asn Val Lys Gly Leu Thr Asn Val Ala Gly Thr Trp Val Ser
100 105 110
Gly Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser Ala His
115 120 125
His Ser Glu Asp Pro Ser
130
<210> 11
<211> 132
<212> PRT
<213> Vicugna pacos
<400> 11
Met Gly Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro
1 5 10 15
Gly Gly Ser Leu Arg Leu Ser Ala Ala Ser Gly Ser Ile Ser Thr Val
20 25 30
Asp Ile Met Gly Trp Tyr Arg Gln Ala Val Gly Lys Gln Arg Glu Leu
35 40 45
Val Ala Val Val Thr Arg Ser Ala Gly Ala Arg Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Val Lys Ser Thr Val Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Leu Thr Asn Arg Gly Tyr Asp Leu Gly Arg Pro Ala Glu Tyr Pro
100 105 110
Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Pro Ser Glu Pro Lys Thr
115 120 125
Pro Lys Pro Gln
130
<210> 12
<211> 135
<212> PRT
<213> Vicugna pacos
<400> 12
Met Gly Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala
1 5 10 15
Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser
20 25 30
Ala Tyr Asp Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu
35 40 45
Phe Val Ala Ala Ile Ser Trp Ser Gly Gly Arg Thr Tyr Tyr Gly Asp
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Leu
65 70 75 80
Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Ala Ala Gly Arg Asn Ile Tyr Phe Ser Asn Ser Pro Leu Ala
100 105 110
Gly Ser Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser Glu
115 120 125
Pro Lys Thr Pro Lys Pro Gln
130 135
<210> 13
<211> 135
<212> PRT
<213> VICUGNA PACOS
<400> 13
Met Gly Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala
1 5 10 15
Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser
20 25 30
Ala Tyr Asp Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu
35 40 45
Phe Val Ala Ala Ile Ser Trp Ser Gly Gly Arg Thr Tyr Tyr Gly Asp
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Leu
65 70 75 80
Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Ala Ala Gly Arg Asn Ile Tyr Phe Ser Asn Ser Pro Leu Ala
100 105 110
Gly Ser Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser Glu
115 120 125
Pro Lys Thr Pro Lys Pro Gln
130 135
<210> 14
<211> 135
<212> PRT
<213> Vicugna pacos
<400> 14
Met Gly Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala
1 5 10 15
Gly Asp Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Pro Ser Phe Ser
20 25 30
Asp Tyr Ala Val Gly Trp Phe Arg Gln Ala Ile Gly Lys Glu Arg Glu
35 40 45
Phe Val Gly Ala Val Ser Trp Thr Ser Thr Glu Thr Ser Tyr Ala Asp
50 55 60
Ala Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr
65 70 75 80
Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr
85 90 95
Thr Cys Ala Ala Thr Arg Thr Tyr Gly Phe Thr Ser Arg Tyr Gln Thr
100 105 110
Asn Tyr Glu Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser Glu
115 120 125
Pro Lys Thr Pro Lys Pro Gln
130 135

Claims (3)

1. a kind of VHH chains of the nano antibody of anti-Her2, it is characterised in that：The amino acid sequence of described VHH chains is SEQ ID NO ：8 to SEQ ID NO：One of amino acid sequence shown in 14.
2. a kind of Her2 nano antibodies, it is characterised in that:It for Her2 epitopes nano antibody, described nano antibody The amino acid sequence of VHH chains is the amino acid sequence described in claim 1.
3. a kind of DNA molecules, it is characterised in that：It encodes the protein being selected from the group：Her2 described in claim 1 Nano antibody VHH chains.
CN201710136524.7A 2017-03-08 2017-03-08 A kind of nano antibody of anti-Her2 Pending CN106866823A (en)

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Cited By (7)

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WO2018233624A1 (en) * 2017-06-20 2018-12-27 和迈生物科技有限公司 Anti-her2 nanobody and coding sequence and use thereof
CN109627319A (en) * 2019-01-11 2019-04-16 北京协同创新研究院 The heavy chain antibody of anti-HER-2 a kind of and its application
CN110950967A (en) * 2019-12-13 2020-04-03 山东民康生物科技有限公司 Anti-human serum albumin nano antibody and IL-2 fusion protein and preparation method thereof
CN114085287A (en) * 2021-11-12 2022-02-25 中国农业科学院北京畜牧兽医研究所 Canine parvovirus nano antibody CPV-VHH-F5 and application thereof
CN114409787A (en) * 2022-02-18 2022-04-29 南京英瀚斯生物科技有限公司 Separation and purification method for HER3 nano antibody
CN116574186A (en) * 2023-07-07 2023-08-11 云南大学 Nanobody capable of specifically binding HER2 and application thereof
WO2024001510A1 (en) * 2022-06-30 2024-01-04 复旦大学 Preparation method for and use of her2 nanobody and conjugate

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Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018233624A1 (en) * 2017-06-20 2018-12-27 和迈生物科技有限公司 Anti-her2 nanobody and coding sequence and use thereof
US11517632B2 (en) 2017-06-20 2022-12-06 Nanomab Technology Limited Anti-Her2 single chain antibody and coding sequence and use thereof
CN109627319B (en) * 2019-01-11 2022-06-21 北京协同创新研究院 anti-HER-2 heavy chain antibody and application thereof
CN109627319A (en) * 2019-01-11 2019-04-16 北京协同创新研究院 The heavy chain antibody of anti-HER-2 a kind of and its application
CN110950967A (en) * 2019-12-13 2020-04-03 山东民康生物科技有限公司 Anti-human serum albumin nano antibody and IL-2 fusion protein and preparation method thereof
CN110950967B (en) * 2019-12-13 2022-05-13 山东民康生物科技有限公司 Anti-human serum albumin nano antibody and IL-2 fusion protein and preparation method thereof
CN114085287A (en) * 2021-11-12 2022-02-25 中国农业科学院北京畜牧兽医研究所 Canine parvovirus nano antibody CPV-VHH-F5 and application thereof
CN114085287B (en) * 2021-11-12 2022-10-25 中国农业科学院北京畜牧兽医研究所 Canine parvovirus nano antibody CPV-VHH-F5 and application thereof
CN114409787A (en) * 2022-02-18 2022-04-29 南京英瀚斯生物科技有限公司 Separation and purification method for HER3 nano antibody
WO2024001510A1 (en) * 2022-06-30 2024-01-04 复旦大学 Preparation method for and use of her2 nanobody and conjugate
WO2024001844A1 (en) * 2022-06-30 2024-01-04 复旦大学 Method for preparing her2 nanobody and conjugate, and use thereof
CN116574186A (en) * 2023-07-07 2023-08-11 云南大学 Nanobody capable of specifically binding HER2 and application thereof
CN116574186B (en) * 2023-07-07 2023-11-28 云南大学 Nanobody capable of specifically binding HER2 and application thereof

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