CN106117359A - AntiCD3 McAb single domain antibody - Google Patents

AntiCD3 McAb single domain antibody Download PDF

Info

Publication number
CN106117359A
CN106117359A CN201610435643.8A CN201610435643A CN106117359A CN 106117359 A CN106117359 A CN 106117359A CN 201610435643 A CN201610435643 A CN 201610435643A CN 106117359 A CN106117359 A CN 106117359A
Authority
CN
China
Prior art keywords
antibody
cell
single domain
domain antibody
anticd3 mcab
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
Application number
CN201610435643.8A
Other languages
Chinese (zh)
Inventor
李庆
王�忠
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Sun Yat Sen University
Original Assignee
Sun Yat Sen University
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Sun Yat Sen University filed Critical Sun Yat Sen University
Priority to CN201610435643.8A priority Critical patent/CN106117359A/en
Publication of CN106117359A publication Critical patent/CN106117359A/en
Pending legal-status Critical Current

Links

Classifications

    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K16/00Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2809Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against the T-cell receptor (TcR)-CD3 complex
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2317/00Immunoglobulins specific features
    • C07K2317/50Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/569Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2317/00Immunoglobulins specific features
    • C07K2317/60Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/626Diabody or triabody

Landscapes

  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The present invention provides a kind of AntiCD3 McAb single domain antibody, and its aminoacid sequence is as shown in SEQ ID NO.1.The present invention further provides the coding nucleotide sequence of this AntiCD3 McAb single domain antibody, the expression vector comprising this nucleotide sequence and host cell.AntiCD3 McAb single domain antibody antagonism people's CD3 ε N end section that the present invention provides, these antibody may be used for the mensuration of such as CD3, and other need the functional protein of CD3, such as bi-specific antibody.

Description

AntiCD3 McAb single domain antibody
Technical field
The present invention relates to AntiCD3 McAb single domain antibody (sdAb) and aminoacid, nucleotide coding sequence.The invention still further relates to bag Expression vector containing this nucleotide coding sequence and host cell.
Background technology
After monoclonal antibody technique in 1975 comes out, the antibody of antigenic specificity has changed biology and medical science Many aspects.Except the research tool as importance, the availability of monoclonal antibody is that the diagnosis of development human diseases is with new Therapy opens road.But, monoclonal antibody has technical restriction and shortcoming in range of application, such as, move based on suckling Thing expression system is costly.
In the past few decades, a series of new technique and method are for improving the performance of conventional antibodies, especially swollen In tumor treatment.Such as, add poisonous compound to existing antibody, be linked to monoclonal antibody including radiosiotope.So And, even up to today, the Clinical practice monoclonal anti scale of construction is the most little.Another method improving tumorcidal efficiency is to make With the antibody of bispecific, immunocyte is guided to tumor cell, thus kills tumor cell.And it is most commonly used that use is anti- T cell is guided to tumor cell by CD3 antibody.It is by binding the surface antigen of CD3 and tumor cell (TAA) simultaneously, this Bi-specific antibody can trigger the oncolysis of T cell mediation.CD3(cluster differentiation 3) φt cell receptor helps In activating cytotoxic T cell.The protein of a kind of complexity that it is made up of four different chains.In mammal, bag Containing CD3 γ chain, CD3 δ chain and two CD3 ε chains.
A lot of bi-specific antibodys use single-chain antibody.Single-chain antibody be from traditional IgG molecule obtain complete Little Fab.Unfortunately, the scFvs yield of bacterial expression is the lowest.Camel and alpaca comprise a kind of unique types Antibody, this kind of antibody deficiency light chain.Because lacking CH1 than conventional antibody, these so-called heavy chain antibodies have relatively low molecule Amount.The heavy chain of this kind of immunoglobulin variable is called for short VHH, to be different from the variable region of heavy chain of classics.Therefore, individual domain VHH is Minimum available intact antigen combines 15 kDa fragments, is referred to as nano antibody.Nano antibody and Fab, Fv or single-chain antibody phase Ratio, has the advantage of some uniquenesses, as more stable, be easier at bacterial expression.
In prior art, anti-cd 3 antibodies form is single and can not meet actual demand, it is therefore necessary to develop more Anti-cd 3 antibodies, thus for producing and design function albumen provides more and selects.
Summary of the invention
One aspect of the present invention relates to a kind of AntiCD3 McAb single domain antibody, and its aminoacid sequence is as shown in SEQ ID NO. 1.
Another aspect of the present invention relates to the nucleotide sequence of the AntiCD3 McAb single domain antibody of a kind of code book invention.A reality Executing in mode, this nucleotide sequence is as shown in SEQ ID NO. 2.
Another aspect of the present invention relates to a kind of expression vector, and it comprises the nucleotide sequence of the present invention.The present invention also relates to And comprise the host cell of this expression vector.In one embodiment, this host cell is escherichia coli.
The invention still further relates to a kind of bi-specific antibody, it comprises arbitrary CD3 single domain antibody of the present invention.
AntiCD3 McAb single domain antibody antagonism people's CD3 ε N-end section that the present invention provides, these antibody may be used for such as The mensuration of CD3, and other need the functional protein of CD3, such as bi-specific antibody.
Accompanying drawing explanation
Fig. 1 is the skeleton drawing of sdAb phage display library.
Fig. 2 shows serum titer experimental result.
Fig. 3. the agarose gel electrophoresis figure of the total serum IgE of separation of lymphocytes.M swimming lane, DNA marker Marker III; 1-3 swimming lane, is isolatable from the total serum IgE of the lymphocyte drawn blood for the first time;4-7 swimming lane, is isolatable from the lymph that second time is drawn blood The total serum IgE of cell;8-10 swimming lane, is isolatable from the total serum IgE of the lymphocyte that third time is drawn blood.
Fig. 4. the V of purificationHThe agarose gel electrophoresis figure of H PCR primer.M swimming lane, DNA marker Marker III;The 1-8 swimming lane, uses 8 V to the purification that primer obtains from the PBMC of blood drawing for the first timeHH PCR primer;9-16 swimming lane, uses 8 V to the purification that primer obtains from the PBMC of second time blood drawingHH PCR primer;17-24 swimming lane, use 8 to primer from the 3rd The V of the purification that the PBMC of secondary blood drawing obtainsHH PCR primer.
Fig. 5. the insertion rate of sdAb phage display library.M13R (-48) and M13F (-47) primer is used to be expanded by PCR Increase 96 library clones randomly selected.Have ~ clone of 1100 bp DNA bands has VHH insert.
Detailed description of the invention
The present invention now will explain in conjunction with following experiment and accompanying drawing further, it should be noted that these experimental examples and accompanying drawing Should not be construed as limitation of the present invention.
1 strategy
The present invention have developed the single domain antibody of AntiCD3 McAb by display technique of bacteriophage, and has carried out following steps: animal is exempted from Epidemic disease and immune response test, the structure of sdAb phage display library, phage display elutriation and FASEBA screening and FACS are tested Card.
2 materials
Antigen protein: CD3-His albumen (MP-5)
Cell antigen system: Jurkat target cell, KPCN compared with control cells
TRIzol® Reagent (Ambion, Cat. No. : 15596-026)
PrimeScriptTM1st Strand cDNA synthetic agent box (Takara, Cat. No.: 6110A)
SfiI enzyme (NEB, Cat. No.: R0123S)
Host Strains:E.coli SS320
Ampicillin, 100 mg/ml
Isopropyl-β-D-thiogalactoside (IPTG), 1 M
PBS: 137 mM NaCl, 2.7 mM KCl, 4.3 mM Na2HPO4, 1.4 mM KH2PO4, pH 7.4
ELISA titer plate (Corning, Cat. No.: 9018)
It is coated buffer: 0.05 M NaHCO3, pH 9.6
Block buffer (PBST): PBS, pH 7.4, add 5% defatted milk powder
Lavation buffer solution: PBS, pH 7.4, add 0.05% Tween 20
M13KO7 helper phage (NEB, Cat. No.: N0315S)
HRP/ anti-M13 monoclonal antibody (GE Healthcare, Cat. No.: 27-9421-01)
Goat-anti camel IgG [HRP] (Novus Biological, Cat. No.: NB7242)
Rabbit anti-camel IgG [HRP] (GenScript)
FACSCalibur (BD Bioscience, San Jose, CA)
Flowjo: Flowjo 7.6.1 Min software
2 × YT:16 g tryptone, 10 g yeast extracts, 5 g NaCl are dissolved in 1 L ddH2O.
OctetRED96 (ForteBio)
Super Streptavidin (SSA) Dip and ReadTM Biosensors (ForteBio)
10 mM glycine-HCl, pH 2.0
3 methods
3.1 animal immunes and immune response test
3.1.1 animal immune
CD3-His immunogen is mixed with adjuvant or PBS and is expelled to yamma (llama).In whole Project Process, animal By immunity five times.Peripheral blood sample is gathered respectively before immunity and after the 4th and the 5th immunity.Gradient separations method is divided Separate out lymphocyte.Cell adds RNAlater and is stored in-80 ° of C.The centrifugal blood adding anticoagulant obtains serum, And it is stored in-80 ° of C.
3.1.2 immune response test
Blood serum sample is used to be evaluated for fixing immunogenic immune response by ELISA.Before have rated immunity and the 4th With the serum after the 5th immunity.Antigen (CD3-His and CD34-Fc) is diluted in being coated in buffer of 4 μ g/ml respectively.Make With the antigen of dilution, titer plate is coated overnight, 4 ° of C.Subsequently, titer plate is washed 3 times with lavation buffer solution, then at 37 ° of C Close 2 hours with Block buffer.Titer plate is washed 4 times again with lavation buffer solution.The serum of a series of dilutions is added plate On, hatch 2 hours at 37 ° of C.Titer plate is washed 4 times subsequently with lavation buffer solution.Add goat-anti camel IgG [HRP] or rabbit resists Camel IgG [HRP], hatches 1 hour at 37 ° of C.After washing, reactant adds tmb substrate and reacts 10 minutes, be subsequently added 1 M HCl terminates reaction.MK3 spectrometer measures every hole absorption value at 450 nm.
The structure of 3.2 sdAb phage display libraries
3.2.1 RNA extracts
From the lymphocyte (see 3.1.1) separated, total serum IgE is extracted according to TRIzol reagent handbook.Gel electrophoresis and Total serum IgE is carried out qualitative and quantitative by OD260/280 method.
3.2.2 RT-PCR and VHH expands
According to PrimeScriptTMThe handbook of 1st Strand cDNA synthetic agent box uses oligo (dT) 20 primer by total RNA reverse transcription is cDNA.Design four justice and two antisense specific degenerate primers expand VHH fragment, introduces twoSfiI restriction site.V is expanded according to GenScript S.O.P. (SOP)HH fragment.
3.2.3 show storehouse extension
Use different primers that amplification is obtained VHH PCR primer.UseSfiI digests this PCR primer and passes through gel-purified. The fragment of gel-purified is inserted in the phagemid vector (Fig. 1) that GenScript is own.Build experimental displaying storehouse to optimize Connect and conversion condition.The connection and the conversion condition that optimize are used to develop actual displaying storehouse.It is dilute that sub-fraction converts cell Release and line on 2 × YT flat board with 100 μ g/ml ampicillin.Bacterium colony is calculated library size. Random picking positive colony also checks order, to assess the quality in library.Remaining converts cell and is scribed in having 100 μ g/ml ammonia benzyls Penicillin and 2% glucose × YT flat board on.Lawn is scraped off from flat board.Fraction cell is used for Library plasmid and separates.Its Remaining addition glycerol, is stored in-80 ° of C standby.
3.3 phage library elutriations
3.3.1 biopanning
The standardization program using GenScript exploitation carries out the elutriation of CD3-His albumen to constructed sdAb phage library.Contain Have 7.4 × 108The big sdAb phage display library of individual clone (CFU) is used for this screening.Library is grown on logarithmic (log) phase, uses M13KO7 helper phage is reclaimed, and trains at the 2 × YT with 100 μ g/ml ampicillin and 50 μ g/ml kanamycin Support in plate and expand overnight at 30 ° of C on agitator.Make phages with PEG/NaCl, and be resuspended in PBS, be stored in- 80°C.For carrying out phage elutriation, microwell plate (Pierce, Prod# 15100) being coated CD3-His, this is by by them Realize in 4 ° of C overnight incubation in PBS (pH 7.0) with 100 μ g/ml.Meanwhile, phage particle is taken off with containing 2% The PBS Block buffer of fat milk powder at room temperature hatches 1 hour to block non-specific binding.After rinsing 3 times with PBS, will Phage particle adds in micropore, and on the oscillator in incubated at room 1 hour.After hatching, by containing 0.05% with PBST( The PBS of Tween-20) rinsing aperture 6 times washes away biting of unconjugated and non-specific binding for 2 times followed in turn by PBS rinsing Thalline.In conjunction with phage be used to immediately infect exponential phases at 37 ° of CE. coli(OD600 is about TG1 cell 0.5) 1 hour.After each panning rounds, infected cell and 10% glycerol are mixed, is subsequently stored in-80 ° of C.Carry out During the elutriation that the next one circulates, by the infection of 10 mlE. coliTG1 cell liquid storage adds 30 ml and contains 200 μ g/ml ammonia In the culture medium of benzylpcnicillin and 2% glucose, and grow to logarithmic (log) phase.Culture reclaims with M13KO7 helper phage, amplification And precipitating phage, screen for next round.Conventional phage display elutriation repeated as described above.
3.3.2 Phage-ELISA
Single output phage clone is grown in 96 deep well plate and screens to confirm CD3-His by Phage-ELISA Specificity is cloned.2 μ g/ml CD3-His are used to be coated 96 hole flat boards (in 4 ° of C overnight incubation in being coated buffer), to contain The PBS having 2% defatted milk powder closes.Every hole adds the phage supernatants from overnight cell culture of about 50 μ l, and at 4 ° C is hatched 2 hours, washs four times with PBST subsequently.Unconjugated sdAb phage by the anti-M13 monoclonal antibody with HRP labelling at 37 ° C is hatched 1 hour and is detected.With PBST washing hole four times again, each hole adds substrate solution.Measure at 450 nm and absorb Value.
3.4 FASEBA screenings and FACS verify
3.4.1 FASEBA screening and affine classification
Amplification exports the DNA of the coding sdAb fragment of phage and is inserted intopTo filter out guide's antibody in FASEBA carrier.? 96 deep well plate are inoculated single FASEBA library clone abduction delivering.Carry out ELISA to screen to separate specific recognition The sdAb of CD3-His albumen, and select the expression culture medium of the affine classification for Octet.At Octet RED96 instrument (ForteBio) dissociation rate screening (Off-rate screening) is carried out on.All tests are all carried out at 30 ° of C.Express training SdAb-SASA albumen in foster base is caught into and is coated the biosensor of BSA and with CD3 albumen at solution (1x PBS, pH 7.4,0.05% Tween-20) in hatch together.The CD3 of one of which concentration (100 nM) is used for dissociation rate classification.Base Line and dissociation steps are only carried out in buffer (1x PBS, pH 7.4,0.05% Tween-20).Use Fortebio data Process software and measure binding kinetics with dynamics data analytical model by 1:1 Langmuir binding pattern.With CD3 with High-affinity interact conjugate screened go out and carry out DNA sequencing.
3.4.2 FACS checking
Combine Jurkat cell to filter out and do not combine the sdAb of KPCN cell, culture supernatants is carried out fluidic cell Analyze.Cell is collected by being centrifuged when Jurkat cell and KPCN cell grow to 70-80% fusion rate.Every hole inoculation about 4 × 105Individual cell, washs 2 times with PBS.In cell, add the culture supernatants of 200 l and at room temperature hatch 1 hour.With After PBS washing, in cell, add the sdAb that antibody combines with detection.After 30 minutes hatch, with PBS washed cell twice, and Cell is resuspended in PBS.FACSCalibur (BD Bioscience, San Jose, CA) and Flowjo software is used to divide The sdAb of analysis cell combines.
4. result
4.1 immune response tests
Fig. 2 shows serum titer experimental result.First time and the potency ratio preimmune serum of second time test sera after immunity Much higher.For the first time and significant difference is not observed between the titer of second time test sera after immunity.
4.2 sdAb phage display library builds
4.2.1 Total RNAs extraction
Isolating about 449 g total serum IgE (Fig. 3) lymphocyte from all acquisition, the total serum IgE of about half is used to build high-quality Library.
4.2.2 RT-PCR and VHH expands
SOP according to GenScript uses 8 primer (x2 antisense primer of 4 sense primers) is carried out RT-PCR.Use difference The product obtained is mixed together and carries out gel-purified (Fig. 4) by primer.Obtain altogether the about 24 pure V of gHH PCR primer. General PCR primer is used for library construction.
4.2.3 library extension
At least 7.4 × 107Individual transformant can obtain from one group of electricity converts, and converts so having carried out 10 times abreast, to obtain Final library.Library size about ~ 7.4 × 108For.Screening according to bacterium colony, insertion rate is 97.92% (94/96) (Fig. 5).One Checked order 72 clones altogether, and 64 clones are positioned at frame.Neither one has identical aminoacid sequence (data are not shown), table Bright sdAb library has the multiformity of height.
4.3 phage library elutriations
Having carried out two-wheeled elutriation and Phage-ELISA, table 1 lists correlative detail.
Table 1. elutriation and the details of Phage-ELISA experiment
Round Input (pfu) Output (pfu) Phage-ELISA positive rate
1st 2.0×1011 3.0×105 ~ 5%
2nd 3.5×1010 9.7×107 ~ 51%
4.4 FASEBA screenings and FACS verify
4.4.1 FASEBA screening and affine classification
The DNA of the coding sdAb fragment of output phage is taken turns in amplification second, and insertspTo be sieved by ELISA in FASEBA carrier Select guide's antibody.32 clones are obtained by FASEBA ELISA screening.These 32 clones carry out DNA sequencing, and carry out with The combination test of CD3, carries out dissociation rate analysis by Octet RED96 subsequently.In conjunction with the result (4.4.2) of FACS, utilize FortteBio data analysis software selects the independent clone of low part of dissociating in curve.Table 2 shows selected sdAb gram Grand dynamics data.
The dynamics data of table 2. sdAb clone
Sample ID Serial number koff(1/s)
C12 SEQ ID NO. 1 6.61E-05
4.4.2 FACS checking
Being verified that by FACS the Jurkat cell of positive culture (from 4.4.1) supernatant combines, KPCN is used as negative right Photo cell system.In conjunction with the result (4.4.1) of affine classification, it is listed in table 3 in conjunction with MFI.
Table 3. culture supernatants and Jurkat cell and the combination MFI of KPCN cell *
Sample C12 NC-sdAb TG1
Jurkat cell 2330.03 115.03 132.03
KPCN cell 72.6 51 54.5
*: the culture supernatants of NC-sdAb (the non-correlation sdAb of same form) is arranged to negative control, the training of TG1 Support thing supernatant and be arranged to ground control.
SEQUENCE LISTING
<110>Zhongshan University
<120>AntiCD3 McAb single domain antibody
<130> 16510CN
<160> 2
<170> PatentIn version 3.5
<210> 1
<211> 142
<212> PRT
<213>artificial sequence
<400> 1
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Met Tyr Arg Pro Asn
20 25 30
Thr Met Gly Trp His Arg Gln Ala Pro Gly Glu Gln Arg Lys Leu Val
35 40 45
Ala Arg Ile Thr Ser Asp Gly Asn Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val His Leu
65 70 75 80
Glu Met Asn Ser Leu Gln Pro Glu Asp Thr Ala Val Tyr Tyr Cys Tyr
85 90 95
Tyr Ser Asp Phe Val Asn Leu Arg Gly Phe Trp Gly Gln Gly Thr Gln
100 105 110
Val Thr Val Ser Ser Ser Ile Ala Thr Met Thr Leu Asp Pro Lys Val
115 120 125
Phe Ala Ser Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
130 135 140
<210> 2
<211> 399
<212> DNA
<213>artificial sequence
<400> 2
caggtgcagc tgcaggagtc tgggggaggc ttggtgcagc ctggggggtc tctgagactc 60
tcctgtgcag cctctggaag catgtataga cccaatacca tgggctggca ccgccaggct 120
ccaggcgagc agcgcaagtt ggtcgcacgc attacaagtg atggtaacac aaactatgca 180
gactccgtga agggccgatt caccatctcc agagacaacg ccaagaacac ggtgcatctc 240
gaaatgaaca gcctacagcc tgaggacacg gccgtctatt actgttacta tagcgacttt 300
gtaaatttaa ggggcttctg gggccagggg acccaggtca ccgtctccag cggccgctac 360
ccgtacgacg ttccggacta cggttccggc cgagcatag 399

Claims (7)

1. an AntiCD3 McAb single domain antibody, its aminoacid sequence is as shown in SEQ ID NO. 1.
2. the nucleotide sequence of the AntiCD3 McAb single domain antibody that a kind encodes described in claim 1.
Nucleotide sequence the most according to claim 2, wherein said nucleotide sequence is as shown in SEQ ID NO. 2.
4. an expression vector, it comprises the nucleotide sequence described in Claims 2 or 3.
5. the host cell of the expression vector comprised described in claim 4.
Host cell the most according to claim 5, wherein said host cell is escherichia coli.
7. a bi-specific antibody, it comprises the aminoacid sequence of the AntiCD3 McAb single domain antibody described in claim 1.
CN201610435643.8A 2016-06-17 2016-06-17 AntiCD3 McAb single domain antibody Pending CN106117359A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610435643.8A CN106117359A (en) 2016-06-17 2016-06-17 AntiCD3 McAb single domain antibody

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610435643.8A CN106117359A (en) 2016-06-17 2016-06-17 AntiCD3 McAb single domain antibody

Publications (1)

Publication Number Publication Date
CN106117359A true CN106117359A (en) 2016-11-16

Family

ID=57470741

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610435643.8A Pending CN106117359A (en) 2016-06-17 2016-06-17 AntiCD3 McAb single domain antibody

Country Status (1)

Country Link
CN (1) CN106117359A (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110218256A (en) * 2018-03-02 2019-09-10 广西医科大学 Nano antibody CD3/Nb29 of AntiCD3 McAb and the preparation method and application thereof
CN110218254A (en) * 2018-03-02 2019-09-10 广西医科大学 Nano antibody CD3/Nb25 of AntiCD3 McAb and the preparation method and application thereof
CN110218253A (en) * 2018-03-02 2019-09-10 广西医科大学 Nano antibody CD3/Nb14 of AntiCD3 McAb and the preparation method and application thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104487587A (en) * 2012-04-20 2015-04-01 新兴产品开发西雅图有限公司 Cd3 binding polypeptides

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104487587A (en) * 2012-04-20 2015-04-01 新兴产品开发西雅图有限公司 Cd3 binding polypeptides

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110218256A (en) * 2018-03-02 2019-09-10 广西医科大学 Nano antibody CD3/Nb29 of AntiCD3 McAb and the preparation method and application thereof
CN110218254A (en) * 2018-03-02 2019-09-10 广西医科大学 Nano antibody CD3/Nb25 of AntiCD3 McAb and the preparation method and application thereof
CN110218253A (en) * 2018-03-02 2019-09-10 广西医科大学 Nano antibody CD3/Nb14 of AntiCD3 McAb and the preparation method and application thereof
CN110218253B (en) * 2018-03-02 2020-12-04 广西医科大学 CD 3-resistant nano antibody CD3/Nb14 and preparation method and application thereof
CN110218254B (en) * 2018-03-02 2020-12-04 广西医科大学 CD 3-resistant nano antibody CD3/Nb25 and preparation method and application thereof
CN110218256B (en) * 2018-03-02 2020-12-08 广西医科大学 CD 3-resistant nano antibody CD3/Nb29 and preparation method and application thereof

Similar Documents

Publication Publication Date Title
DK3124973T3 (en) AFLATOXIN-NANOBODY IMMUNO ABSORBENT AND IMMUNO-EFFICIENCY COLUMN AND PREPARATION AND METHOD OF USING IT
JP6391574B2 (en) Method for producing an antibody molecule having interspecific intra-target cross-reactivity
CN109942708B (en) anti-BCMA single domain antibody and application thereof
CN106164091A (en) Tcr library
CN106084046A (en) AntiCD3 McAb single domain antibody
CN107955071A (en) Human-derived anti-human CD47 antibody and its encoding gene and application
CN106084048A (en) AntiCD3 McAb single domain antibody
CN106892980B (en) anti-VEGFR 2 monoclonal antibody and application thereof
EP2761064B1 (en) Molecular display method
CN114262377B (en) Preparation method of anti-human CD70 nano antibody for blocking binding of CD70 and ligand CD27 thereof and coding sequence thereof
CN112028997B (en) anti-CEACAM 5 nano antibody
CN106604932A (en) Immune-stimulating monoclonal antibodies against human interleukin-2
Eteshola Isolation of scFv fragments specific for monokine induced by interferon-gamma (MIG) using phage display
JP7566352B2 (en) Fully human antibodies targeting CD19 and their applications
CN106117359A (en) AntiCD3 McAb single domain antibody
EP3271393A1 (en) High throughput monoclonal antibody generation by b cell panning and proliferation
CN110156895A (en) A kind of anti-PD-L1 antibody or its functional fragment and application thereof
CN106146661A (en) AntiCD3 McAb single domain antibody
EP3394256B1 (en) A method of generating an antibody naïve library, said library and application(s) thereof
CN110903393B (en) Polypeptide capable of binding IL6R alpha and application thereof
CN106117358A (en) AntiCD3 McAb single domain antibody
CN106146662A (en) AntiCD3 McAb single domain antibody
CN106084049A (en) AntiCD3 McAb single domain antibody
CN106084047A (en) AntiCD3 McAb single domain antibody
Link et al. Selection of phage-displayed anti-guinea pig C5 or C5a antibodies and their application in xenotransplantation

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20161116

WD01 Invention patent application deemed withdrawn after publication