CN108341877A - A kind of anti-activation labile factor monoclonal antibody and its preparation method and application - Google Patents
A kind of anti-activation labile factor monoclonal antibody and its preparation method and application Download PDFInfo
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- CN108341877A CN108341877A CN201710050407.9A CN201710050407A CN108341877A CN 108341877 A CN108341877 A CN 108341877A CN 201710050407 A CN201710050407 A CN 201710050407A CN 108341877 A CN108341877 A CN 108341877A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/36—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against blood coagulation factors
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/86—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood coagulating time or factors, or their receptors
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/745—Assays involving non-enzymic blood coagulation factors
- G01N2333/7456—Factor V
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Abstract
The invention discloses a kind of anti-activation labile factor monoclonal antibodies, it is characterised in that：The amino acid sequence of its light chain such as SEQ ID NO：Shown in 3, the amino acid sequence such as SEQ ID NO of heavy chain：Shown in 4.The invention also discloses the preparation method of aforementioned monoclonal monomer and purposes.Anti-activation labile factor monoclonal antibody of the present invention and the binding ability of activated clotting factor V are strong, can be used for the detection of activated clotting factor V.
Description
Technical field
The invention belongs to biotechnologies, and in particular to a kind of anti-activation labile factor monoclonal antibody and its preparation
Method and purposes.
Background technology
Labile factor (factor V, FV) is the single chain glycoprotein that molecular weight is 333KD, almost with nothing under physiological conditions
Active state is present in blood plasma.Under the action of fibrin ferment, plasma F V be changed into the activation blood coagulation with blood coagulation activity because
Sub- V (ctivated factor V, FVa).FVa in blood plasma is an important co-factor in coagulation process, to coagulation process
There are vital regulating and controlling effect, mechanism as shown in Figure 1.In coagulation process, FVa and FXa, calcium ion and the common structure of phosphatide
At prothrombin complex, the activation efficiency of factor can be made to increase by 300,000 times.FVa can pass through the PROTEIN C of activation again
(activated protein C, APC) is cracked and is inactivated, to directly or indirectly participate in coagulation process by APC approach.
In short, FVa is in the crosspoint for promoting coagulation pathway and anticoagulation approach in coagulation process, to maintaining coagulation homeostasis to play to pass
Important role.
FV is not only played an important role in terms of coagulation homeostasis, but also the active height of plasma F V content, FV is also and disease
There is correlation in disease.Studies have shown that FV is abnormal in vivo, according to the difference of generation mechanism, heredity FV can be caused to lack
Disease causes bleeding；Or lead to activated protein C resistance (APCR), cause thrombus.In addition, the active measurement of plasma coagulation factors V,
It is also used as judging the serious hepatitis state of an illness and observes a useful indicators of its prognosis.
But still lack the method for effective quantitative detection activation FV at present, develop anti-activation FV monoclonal antibodies for
Establish high specificity, high sensitivity FVa activity test methods, it is significant with auxiliary diagnosis FV abnormal diseases.It has no at present
The preparation of anti-activation FV monoclonal antibodies is reported.
Invention content
The purpose of the present invention is to provide a kind of anti-activation labile factor monoclonal antibodies.
Anti-activation labile factor monoclonal antibody of the present invention, the amino acid sequence such as SEQ ID NO of light chain variable region：3
It is shown, the amino acid sequence such as SEQ ID NO of heavy chain variable region：Shown in 4.
The present invention encodes the genetic fragment of aforementioned monoclonal antibody, the nucleotide sequence such as SEQ ID of light chain variable region
NO：Shown in 1, the nucleotide sequence SEQ ID NO of heavy chain variable region：Shown in 2.
Recombinant vector of the present invention, it contains forementioned gene segment.Wherein, the carrier is recombination pcDNA3.1/ZEO (+)
Carrier.
Transgenic cell of the present invention, it contains recombinant vector above-mentioned.Wherein, the transgenic cell is transgenosis CHO-
K1 cells.
The method that the present invention prepares monoclonal antibody above-mentioned, it is characterised in that：Steps are as follows：
(1) transgenic cell above-mentioned is taken, is cultivated；
(2) culture solution supernatant is taken, is purified, you can.
Wherein, the method for the purifying is to use Hi Trap Protein A affinity column chromatographic purifyings.
The present invention also provides purposes of the aforementioned monoclonal antibody in the reagent for preparing detection activated clotting factor V.
Anti-activation labile factor monoclonal antibody has been prepared in the present invention, has high-affinity to FVa antigens, is used for
People source FVa high specificities are isolated and purified and detected, the reagent of detection activated clotting factor V is can be used as, are used for activated clotting factor V
Detection, application prospect is good.
Obviously, the above according to the present invention is not being departed from according to the ordinary technical knowledge and customary means of this field
Under the premise of the above-mentioned basic fundamental thought of the present invention, the modification, replacement or change of other diversified forms can also be made.
The specific implementation mode of form by the following examples remakes further specifically the above of the present invention
It is bright.But the range that this should not be interpreted as to the above-mentioned theme of the present invention is only limitted to example below.It is all to be based on the above of the present invention
The technology realized all belongs to the scope of the present invention.
Description of the drawings
Fig. 1 is ELISA method detection bacteriophage affinity result figure.
Fig. 2 is to carry out gradient concentration compatibility detection figure with ELISA method to obtained monoclonal antibody purification.
Specific implementation mode
It is described further below with embodiment, but the present invention is not limited to these embodiments.
It is prepared by the monoclonal antibody of 1 anti-activation FV of the present invention of embodiment
One, the full people source FVa single-chain antibodies of Phage Antibody Library
Using FVa recombinant proteins (HTI Products) as the full Large human naive scFv phage library of antigen selection.By FVa antigens
It is diluted to coating dilution (0.05mol/L sodium carbonate-bicarbonates, pH 9.6) and is interpreted into 10ug/mL, enzyme linked immunological plate is per hole
100ul is added, sets 4 DEG C of refrigerator coatings overnight.Confining liquid is mixed by PBST (PBS, 0.05%Tween 20) and 1%BSA,
Add 200ul per hole, closes 1 hour.PBST is washed 5 times, each 3min, and the Fab-based phage display libraries in full people source are added
(deriving from Shanghai Bo Yu biotechnologies company phage display library), is incubated overnight at room temperature.Liquid in immune plate is abandoned, PBST is washed
It washs 5 times, each 3min, the bacteriophage combined closely with FVa antigens is eluted with eluent (50mM HCl, 500mM NaCl)
30min, the eluent are neutralized with isometric 1M Tris base.Escherichia coli XL-1Blue bacterial strains and helper phage is added
Body is incubated overnight, and enrichment has the specific bacteriophage of affinity with FVa.This screening process is repeated twice.
ELISA method detection bacteriophage affinity picks out the strongest preceding 10 plants of bacteriophages of affinity.
ELISA method：FVa antigens coating diluted to 1ug/mL, it is diluted that 100ul is added per hole for enzyme linked immunological plate
Antigen sets 4 DEG C of refrigerator overnights.Confining liquid 150ul is added per hole, sets 37 DEG C of closing 40min.Cleaning solution 200ul is added per hole to wash
It washs 3 times, it is every all over 3min.Monoclonal recombinant phage to be measured, 37 DEG C of incubation 2h are added.It is washed 3 times, every time with cleaning solution
3min.Goat-anti people-HRP (dilution ratios 1 are added:2000) working solution after 37 DEG C are incubated 1h, adds 200ul cleaning solutions to wash 3 per hole
It is secondary, each 3min.Tmb substrate the solution 100ul, 37 DEG C of incubation 20min of Extemporaneous are added in each reacting hole.It is added per hole
2M sulfuric acid 50ul are added in each reacting hole, terminate reaction, with measurement OD values under microplate reader 450nm wavelength.
ELISA testing results are as shown in Figure 1,131 sample OD450nmValue both greater than 0.6, wherein there is 57 sample OD450nm
More than 2；And only 61 sample OD450nmValue is less than 0.6, that is, 131 bacteriophage monoclonal antibodies generally have with FVa antigens
There is higher positive Percentage bound, and it is in strong positive there are 57 sample antigens to combine；61 sample antigens, which combine, to be negative.
Two, carrier for expression of eukaryon is built
The strongest preceding 10 kinds of bacteriophages of affine competitiveness are will be singled out, respectively using plasmid gene as template, primer is added,
PCR amplification variable region VLGene；Again with human antibody VLAnd CLFor template, L chain genes are amplified, are cloned into LPG3 carriers；
Using bacteriophage monoclonal antibody DNA as template, primer is added, PCR amplification goes out VHGene is cloned into LPG4 carriers
In；And using people source cDNA as template, primer is added, expands CHArea's gene is cloned into LPG4 carriers.
By screening, obtain the nucleotide sequence (SEQ ID NO.1) for expressing antibody 1C11 light chain variable regions of the present invention and
The nucleotide (SEQ ID NO.2) of heavy chain variable region.
Antibody light chain nucleotide sequence of the present invention (nucleotides sequence of variable region is classified as SEQ ID NO.1) is taken, with Hind III
It is connected on plasmid pcDNA3.1/ZEO (+) (Invitrogen Products) with the mode of I double digestions of EcoR, build eukaryon
Expression vector.
Heavy chain of antibody nucleotide sequence of the present invention (nucleotides sequence of variable region is classified as SEQ ID NO.2) is taken, with Hind III
It is connected on plasmid pcDNA3.1/ZEO (+) (Invitrogen Products) with the mode of I double digestions of EcoR, builds eukaryon table
Up to carrier.
By verification, the present invention builds to have obtained the full people containing segment shown in SEQ ID NO.1 or SEQ ID NO.2
Source carrier for expression of eukaryon.
Wherein, light chain nucleotide sequence of the present invention is made of the sequence of SEQ ID NO.1 and constant region, heavy chain nucleotide sequence
Row are made of the sequence of SEQ ID NO.2 and constant region, and sequence can be synthesized directly, be then added to pcDNA3.1/ZEO (+)
In, form recombinant eukaryon expression vector.
Three, the expression and purifying of antibody
Chinese hamster ovary cell (Chinese hamster ovary celI) is accurate when expressing human antibody and less secretion intrinsic protein, because
This present invention uses expression vector of the Chinese hamster ovary celI as monoclonal antibody.
3×105CHO-k1 cell inoculations are in tissue culture dishes, and in common DMEM culture mediums, (GIBCO companies produce cell
Product) in culture to 85% fusion when start to transfect【The method of transfection：By 3x105CHO-K1 cell inoculations are to 3.5cm tissue cultures
It in ware, is transfected when cell confluency is to 90%-95%, transfection process is：The recombinant plasmid 10ug of above-mentioned structure is taken to use
Lipofectamine2000Reagent kits (Invitrogen companies) are transfected by its specification】, 37 DEG C, 5%CO2
After incubator is incubated for 24 hours, Selective agar medium, i.e. G418 containing 600ug/ml (Invitrogen Products) and 250ug/ are changed into
The DMEM culture mediums of ml Zeocin (Invitrogen Products) are screened；Continue after expanding culture, with Hi Trap
Protein A affinity columns (GE Products) purify culture solution, are eluted, are purified with 0.1M sodium acetates (PH 3.0)
Monoclonal antibody 1C11 of the present invention.
By sequencing, the amino acid sequence such as SEQ ID NO.3 institutes of the light chain variable region of monoclonal antibody 1C11 of the present invention
Show, the amino acid sequence of heavy chain variable region is as shown in SEQ ID NO.4.
Illustrate beneficial effects of the present invention with the mode of experimental example below：
The antigen-binding activity of 1 monoclonal antibody of the present invention of experimental example measures
1, experimental method
The monoclonal antibody purification 1C11 that Example 1 is prepared carries out gradient concentration compatibility inspection with ELISA method
It surveys.
ELISA method：FVa antigens coating diluted to 1ug/mL, it is diluted that 100ul is added per hole for enzyme linked immunological plate
Antigen sets 4 DEG C of refrigerator overnights.Confining liquid 150ul is added per hole, sets 37 DEG C of closing 40min.Cleaning solution 200ul is added per hole to wash
It washs 3 times, it is every all over 3min.Monoclonal recombinant phage to be measured, 37 DEG C of incubation 2h are added.It is washed 3 times, every time with cleaning solution
3min.Goat-anti people-HRP (dilution ratios 1 are added:2000) working solution after 37 DEG C are incubated 1h, adds 200ul cleaning solutions to wash 3 per hole
It is secondary, each 3min.Tmb substrate the solution 100ul, 37 DEG C of incubation 20min of Extemporaneous are added in each reacting hole.It is added per hole
2M sulfuric acid 50ul are added in each reacting hole, terminate reaction, with measurement OD values under microplate reader 450nm wavelength.
2, experimental result
Result figure 2.
From Figure 2 it can be seen that monoclonal antibody 1C11 of the present invention and activated clotting factor V (FVa) antigen-binding activity is very strong,
EC50 values are 0.03732.
In Fig. 2, remaining 4 kinds of antibody such as 2H11 are also using the antibody of Phage Antibody Library method screening, still
Their combination activity is significantly lower than monoclonal antibody 1C11 of the present invention.
To sum up, anti-activation labile factor monoclonal antibody of the invention has high-affinity to people's FVa antigens；In addition,
The preparation method of the anti-FV monoclonal antibodies in full people source of the present invention is simple, can be suitable for large-scale production, and application prospect is good.
SEQUENCE LISTING
<110>Wu, sword wave
Poplar, swallow
<120>A kind of anti-activation labile factor monoclonal antibody and its preparation method and application
<130> GY421-17P1042
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 336
<212> DNA
<213>Light chain variable region（VL）Nucleotide sequence
<400> 1
gatattgtga tgactcagtc tccactctcc ctgcccgtca cccttggaca ggcggcctcc 60
atctcctgca ggtctagtca aagcctcgta tacagtgatg gaaacaccta cttgaattgg 120
tttcagcaga ggccaggcca atctccaagg cgcctaattt ataaggtttc taaccgggac 180
tctggggtcc cagacagatt cagcggcagt gggtcaggca ctgaattcac actgagagtc 240
agcagggtgg aggctgagga tgttggtgtt tattactgca tgcaagctac acactggcct 300
ccgacgttcg gccaagggac cagggtggaa atcaaa 336
<210> 2
<211> 351
<212> DNA
<213>Heavy chain variable region（VH）Nucleotide sequence
<400> 2
caggtccagc tggtgcagtc tggagcagag gtgaaaaagc ccggggagtc tctgaagatc 60
tcctgtaagg gttctggata tagctttacc acctactgga tcgcctgggt gcgccagatg 120
cccgggaaag gcctggagtg gatggggatc atctatcctg gtgactctga taccagatac 180
agcccgtcct tccaaggcca ggtcaccatc tcagccgaca agtccatcag aaccgcctac 240
ctgcagtgga gcagcctgaa ggactcggac accgccatgt attactgtgc gagacttgat 300
gtctacggtc tggacgtctg gggccaaggg accacggtca ccgtctcctc a 351
<210> 3
<211> 1008
<212> PRT
<213>The amino acid sequence of light chain variable region
<400> 3
Gly Ala Thr Ala Thr Thr Gly Thr Gly Ala Thr Gly Ala Cys Thr Cys
1 5 10 15
Ala Gly Thr Cys Thr Cys Cys Ala Cys Thr Cys Thr Cys Cys Cys Thr
20 25 30
Gly Cys Cys Cys Gly Thr Cys Ala Cys Cys Cys Thr Thr Gly Gly Ala
35 40 45
Cys Ala Gly Gly Cys Gly Gly Cys Cys Thr Cys Cys Ala Thr Cys Thr
50 55 60
Cys Cys Thr Gly Cys Ala Gly Gly Thr Cys Thr Ala Gly Thr Cys Ala
65 70 75 80
Ala Ala Gly Cys Cys Thr Cys Gly Thr Ala Thr Ala Cys Ala Gly Thr
85 90 95
Gly Ala Thr Gly Gly Ala Ala Ala Cys Ala Cys Cys Thr Ala Cys Thr
100 105 110
Thr Gly Ala Ala Thr Thr Gly Gly Thr Thr Thr Cys Ala Gly Cys Ala
115 120 125
Gly Ala Gly Gly Cys Cys Ala Gly Gly Cys Cys Ala Ala Thr Cys Thr
130 135 140
Cys Cys Ala Ala Gly Gly Cys Gly Cys Cys Thr Ala Ala Thr Thr Thr
145 150 155 160
Ala Thr Ala Ala Gly Gly Thr Thr Thr Cys Thr Ala Ala Cys Cys Gly
165 170 175
Gly Gly Ala Cys Thr Cys Thr Gly Gly Gly Gly Thr Cys Cys Cys Ala
180 185 190
Gly Ala Cys Ala Gly Ala Thr Thr Cys Ala Gly Cys Gly Gly Cys Ala
195 200 205
Gly Thr Gly Gly Gly Thr Cys Ala Gly Gly Cys Ala Cys Thr Gly Ala
210 215 220
Ala Thr Thr Cys Ala Cys Ala Cys Thr Gly Ala Gly Ala Gly Thr Cys
225 230 235 240
Ala Gly Cys Ala Gly Gly Gly Thr Gly Gly Ala Gly Gly Cys Thr Gly
245 250 255
Ala Gly Gly Ala Thr Gly Thr Thr Gly Gly Thr Gly Thr Thr Thr Ala
260 265 270
Thr Thr Ala Cys Thr Gly Cys Ala Thr Gly Cys Ala Ala Gly Cys Thr
275 280 285
Ala Cys Ala Cys Ala Cys Thr Gly Gly Cys Cys Thr Cys Cys Gly Ala
290 295 300
Cys Gly Thr Thr Cys Gly Gly Cys Cys Ala Ala Gly Gly Gly Ala Cys
305 310 315 320
Cys Ala Gly Gly Gly Thr Gly Gly Ala Ala Ala Thr Cys Ala Ala Ala
325 330 335
<210> 4
<211> 1053
<212> PRT
<213>Heavy chain variable region（VH）Amino acid sequence
<400> 4
Cys Ala Gly Gly Thr Cys Cys Ala Gly Cys Thr Gly Gly Thr Gly Cys
1 5 10 15
Ala Gly Thr Cys Thr Gly Gly Ala Gly Cys Ala Gly Ala Gly Gly Thr
20 25 30
Gly Ala Ala Ala Ala Ala Gly Cys Cys Cys Gly Gly Gly Gly Ala Gly
35 40 45
Thr Cys Thr Cys Thr Gly Ala Ala Gly Ala Thr Cys Thr Cys Cys Thr
50 55 60
Gly Thr Ala Ala Gly Gly Gly Thr Thr Cys Thr Gly Gly Ala Thr Ala
65 70 75 80
Thr Ala Gly Cys Thr Thr Thr Ala Cys Cys Ala Cys Cys Thr Ala Cys
85 90 95
Thr Gly Gly Ala Thr Cys Gly Cys Cys Thr Gly Gly Gly Thr Gly Cys
100 105 110
Gly Cys Cys Ala Gly Ala Thr Gly Cys Cys Cys Gly Gly Gly Ala Ala
115 120 125
Ala Gly Gly Cys Cys Thr Gly Gly Ala Gly Thr Gly Gly Ala Thr Gly
130 135 140
Gly Gly Gly Ala Thr Cys Ala Thr Cys Thr Ala Thr Cys Cys Thr Gly
145 150 155 160
Gly Thr Gly Ala Cys Thr Cys Thr Gly Ala Thr Ala Cys Cys Ala Gly
165 170 175
Ala Thr Ala Cys Ala Gly Cys Cys Cys Gly Thr Cys Cys Thr Thr Cys
180 185 190
Cys Ala Ala Gly Gly Cys Cys Ala Gly Gly Thr Cys Ala Cys Cys Ala
195 200 205
Thr Cys Thr Cys Ala Gly Cys Cys Gly Ala Cys Ala Ala Gly Thr Cys
210 215 220
Cys Ala Thr Cys Ala Gly Ala Ala Cys Cys Gly Cys Cys Thr Ala Cys
225 230 235 240
Cys Thr Gly Cys Ala Gly Thr Gly Gly Ala Gly Cys Ala Gly Cys Cys
245 250 255
Thr Gly Ala Ala Gly Gly Ala Cys Thr Cys Gly Gly Ala Cys Ala Cys
260 265 270
Cys Gly Cys Cys Ala Thr Gly Thr Ala Thr Thr Ala Cys Thr Gly Thr
275 280 285
Gly Cys Gly Ala Gly Ala Cys Thr Thr Gly Ala Thr Gly Thr Cys Thr
290 295 300
Ala Cys Gly Gly Thr Cys Thr Gly Gly Ala Cys Gly Thr Cys Thr Gly
305 310 315 320
Gly Gly Gly Cys Cys Ala Ala Gly Gly Gly Ala Cys Cys Ala Cys Gly
325 330 335
Gly Thr Cys Ala Cys Cys Gly Thr Cys Thr Cys Cys Thr Cys Ala
340 345 350
Claims (9)
1. a kind of anti-activation labile factor monoclonal antibody, it is characterised in that：The amino acid sequence of its light chain variable region such as SEQ
ID NO：Shown in 3, the amino acid sequence such as SEQ ID NO of heavy chain variable region：Shown in 4.
2. a kind of genetic fragment of coding monoclonal antibody described in claim 1, it is characterised in that：The core of its light chain variable region
Nucleotide sequence such as SEQ ID NO：Shown in 1, the nucleotide sequence SEQ ID NO of heavy chain variable region：Shown in 2.
3. a kind of recombinant vector, it is characterised in that：It contains genetic fragment described in claim 2.
4. recombinant vector according to claim 3, it is characterised in that：The carrier is that recombination pcDNA3.1/ZEO (+) is carried
Body.
5. a kind of transgenic cell, it is characterised in that：It contains the recombinant vector described in claim 3 or 4.
6. the transgenic cell stated according to claim 5, it is characterised in that：The transgenic cell is that transgenosis CHO-K1 is thin
Born of the same parents.
7. a kind of method preparing monoclonal antibody described in claim 1, it is characterised in that：Steps are as follows：
(1) transgenic cell described in claim 5 or 6 is taken, is cultivated；
(2) culture solution supernatant is taken, is purified, you can.
8. according to the method described in claim 7, it is characterized in that：In step (2), the method for the purifying is to use Hi
Trap Protein A affinity column chromatographic purifyings.
9. purposes of the monoclonal antibody described in claims 1 or 2 in the reagent for preparing detection activated clotting factor V.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710050407.9A CN108341877B (en) | 2017-01-23 | 2017-01-23 | Activated blood coagulation factor V resistant monoclonal antibody and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710050407.9A CN108341877B (en) | 2017-01-23 | 2017-01-23 | Activated blood coagulation factor V resistant monoclonal antibody and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108341877A true CN108341877A (en) | 2018-07-31 |
| CN108341877B CN108341877B (en) | 2021-04-30 |
Family
ID=62974324
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710050407.9A Active CN108341877B (en) | 2017-01-23 | 2017-01-23 | Activated blood coagulation factor V resistant monoclonal antibody and preparation method and application thereof |
Country Status (1)
| Country | Link |
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| CN (1) | CN108341877B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0670792A (en) * | 1992-08-25 | 1994-03-15 | Tosoh Corp | Monoclonal antibody reactive with protein recognizing phosphatidyl serine |
| WO2008067056A2 (en) * | 2006-10-16 | 2008-06-05 | Novelmed Therapeutics, Inc. | Method of inhibiting coagulation with human anti-factor va antibodies and use thereof |
| CN104203980A (en) * | 2012-02-22 | 2014-12-10 | 瑞泽恩制药公司 | Anti-big-endothelin-1 (big-et-1) antibodies and uses thereof |
-
2017
- 2017-01-23 CN CN201710050407.9A patent/CN108341877B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0670792A (en) * | 1992-08-25 | 1994-03-15 | Tosoh Corp | Monoclonal antibody reactive with protein recognizing phosphatidyl serine |
| WO2008067056A2 (en) * | 2006-10-16 | 2008-06-05 | Novelmed Therapeutics, Inc. | Method of inhibiting coagulation with human anti-factor va antibodies and use thereof |
| CN104203980A (en) * | 2012-02-22 | 2014-12-10 | 瑞泽恩制药公司 | Anti-big-endothelin-1 (big-et-1) antibodies and uses thereof |
Non-Patent Citations (2)
| Title |
|---|
| 柳胜华: "人血浆凝血因子V的纯化和单克隆抗体细胞株的建立", 《中国优秀硕士学位论文全文数据库》 * |
| 谢飞等: "人血浆凝血因子V双抗体夹心ELISA测定", 《上海医学检验杂志》 * |
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| CN108341877B (en) | 2021-04-30 |
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