CN107051396A - A kind of immune absorption material for pinpointing fixed protein A and preparation method thereof - Google Patents
A kind of immune absorption material for pinpointing fixed protein A and preparation method thereof Download PDFInfo
- Publication number
- CN107051396A CN107051396A CN201611251363.8A CN201611251363A CN107051396A CN 107051396 A CN107051396 A CN 107051396A CN 201611251363 A CN201611251363 A CN 201611251363A CN 107051396 A CN107051396 A CN 107051396A
- Authority
- CN
- China
- Prior art keywords
- recombinant protein
- albumin
- protein
- agarose gel
- reaction
- 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
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
- B01J20/24—Naturally occurring macromolecular compounds, e.g. humic acids or their derivatives
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/38—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving specific interaction not covered by one or more of groups B01D15/265 - B01D15/36
- B01D15/3804—Affinity chromatography
- B01D15/3809—Affinity chromatography of the antigen-antibody type, e.g. protein A, G, L chromatography
Landscapes
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
Summary：The invention belongs to bio-separation engineering field, it is related to a kind of for protein A immunoadsorption material of blood purification and preparation method thereof.Disclose the boiomacromolecule separation material that agarose gel microsphere and recombinant protein A are coupled, the material is using Ago-Gel as carrier, after being activated through allyl glycidyl ether, by with recombinant protein A aminoterminal（C-terminal）Sulfydryl coupling on cysteine, is finally blocked, and realizes and the C-terminal fixed point of albumin A is fixed on agarose gel microsphere, obtain a kind of immune absorption material of albumin A high absorption property.The advantage of the material is：Recombinant protein A structure on agarose gel microsphere is homogeneous, orderly fixation, sorbing material is improved the adsorption capacity of antibody, can be applied to clinically immunoadsorption therapy.
Description
Technical field
The invention belongs to biomaterial for medical purpose, relate particularly to for blood purification protein A immunoadsorption material and
Its preparation method.
Background technology
Even in today that medical science is flourishing, the mankind still have many refractory diseases not capture, intractable LADA
Disease is exactly one of which.Autoimmune disease pathogenesis is complicated, can involve whole body multiple organ multisystem, traditional treatment side
Case is based on glucocorticoid and cell toxicity medicament, but therapeutic effect is not good always, be difficult at present it is tackling, threaten the mankind to be good for
One of health and the principal disease of life.
Immuno absorbence therapy is a kind of new technology grown up nearly more than ten years, and it will have specific affine to pathogenic antibody
The material of power is combined as aglucon with carrier, so as to prepare adsorption column, is optionally removed causing a disease in blood samples of patients and is resisted
Body, so as to reach purification blood, alleviate the purpose of illness.At present, immuno absorbence therapy is in organ transplant rejection, kidney
There is significant curative effect in terms of disease, the nervous system disease, blood disease and angiocardiopathy, be increasingly subject to the extensive pass of medical field
Note.
Albumin A is a kind of protein on some aureus cell walls, and its molecular weight is about 42kD, its amino
Contain 5 highly similar immunoglobulin Fc section lands in end（EDABC）, can be with the antibody in human plasma and its immune suction
The Fc section specific bonds of addendum.Carrier-SPA compound immuno absorbences are prepared into if albumin A is fixed on a certain carrier
Post, when human plasma is by the adsorption column, antibody and its immune complex in blood plasma will be adsorbed specifically, some
Disease and symptom caused by antibody and immune complex, such as systemic loupus erythematosus, autoimmune disease, organ transplant,
Malignant tumour etc. can obtain medical treatment and alleviate.The key of immuno absorbence therapy is the conjunction of immune absorption material in immunoabsorbent column
Into so synthetic technology albumin A being fixed on carrier is most important.At present, on protein A immunoadsorption material synthesis side
The patent of method is a lot（CN1367181A, CN1365853A, CN101069751A, CN101190409A, CN101185878A）, this
A little methods generally are coupled to reach fixed purpose using the amino on albumin A surface with active carrier.Due to the ammonia on albumin A surface
Base is a lot, the different parts of albumin A is distributed in, so that albumin A can be randomly with different parts and carrier conjugation, it is impossible to ensure egg
White A activated centre fully exposes, and reduces its adsorption capacity, synthesizes obtained sorbing material and the adsorption capacity of antibody is all existed
20mg/mL or so.Because adsorption column is not high to the adsorption capacity of antibody, it is impossible to which disposable perfusion reaches the mesh for removing pathogenic antibody
, which prevent the extensive use of Protein A immunoadsorption therapy clinically.
The content of the invention
It is an object of the invention to provide a kind of protein A immunoadsorption material of high absorption property.
It is a further object of the present invention to provide a kind of preparation method of new protein A immunoadsorption material, this method is by egg
White A specific site（C-terminal）It is fixed on agarose gel microsphere carrier, prepares the adsorbent of high protein A fixed amounts, it is right
The adsorption capacity of antibody is significantly improved.
The technical scheme is that：
Protein immunization sorbing material, is the high polymer material that agarose gel microsphere is coupled with recombinant protein A, and it has following
Chemical constitution：
Wherein：
Represent agarose gel microsphere；RSPA represents recombinant protein A；（N）Represent the C-terminal key of recombinant protein A
Close on agarose gel microsphere.
Agarose gel microsphere is a kind of natural polysaccharide, and containing abundant hydroxyl, it may participate in a variety of chemical reaction conversions
For active group, then it is fixed on albumin A reaction on agarose gel microsphere carrier.The present invention basic conception be：From
Activating reagent allyl glycidyl ether is as the reagent of activated sepharose microballoon, and the epoxide group of the reagent can be with agar
Hydroxyl reaction on sugared gel micro-ball, prepares the activated carrier with alkenyl.Alkenyl can exist with the sulfydryl on recombinant protein A
Reacted under ultraviolet wavelength 305nm, so that recombinant protein A specific site is fixed on agarose gel microsphere.What it was synthesized
Detailed process is as follows：
1st, allyl glycidyl ether and agarose gel microsphere are anti-in 40 ~ 60 DEG C in 10 ~ 12mol/L alkaline aqueous solution
It is 20% ~ 40% to answer 4 ~ 8 hours Ago-Gels volume ratio shared in reaction solution；
2nd, after step a reactions terminate, reaction product is filtered, with distilled water flushing to neutrality, the activation fine jade with double bond is obtained
Sepharose microballoon；
3rd, the product obtained by step b is added in phosphate buffer solution, and control ph adds recombinant protein A in the range of 6 ~ 8,
Ultraviolet wavelength 305nm irradiations are lower to react.Reaction temperature is 20 ~ 25 DEG C, and the reaction time is 10 ~ 16 hours.
4th, products therefrom adds mercaptoethanol and carries out end-block reaction in step c, the concentration of capping reagent for 0.1 ~
0.5mol/L, pH value is 6 ~ 8, and the reaction time is 4 ~ 6 hours.
Reaction equation is as follows：
Wherein,Represent agarose gel microsphere；RSPA represents recombinant protein A；（C）Represent recombinant protein A
C-terminal is bonded on agarose gel microsphere.
Albumin A is a kind of protein on some aureus cell walls, and its molecular weight is about 42kD, its amino
Contain 5 highly similar immunoglobulin Fc section lands in end（EDABC）.B lands are made up of three helical regions, respectively
For the spirals of α 1（Lys7—His8）, the spirals of α 2（Glu25—Asp36）, the spirals of α 3（Ser41—Ala54）, combined only with antibody Fc section
There are the spirals of α 1 and the spirals of α 2, positioned at c-terminus（C-terminal）The spirals of α 3 not with antibody binding.Because land does not have cysteine residual
Base, we are inserted into cysteine the C-terminal of land by the method for genetic engineering, using the sulfydryl on cysteine with
Albumin A is fixed on agarose gel microsphere by the characteristic reaction of alkenyl, so that the C-terminal of albumin A is fully exposed in agarose
On gel micro-ball, the Fc lands of albumin A are fully exposed, so obtained protein A immunoadsorption material has high inhale to antibody
Attached performance.
Due to spatial obstacle, albumin A is with after the agarose gel microsphere reaction with alkenyl, also having some unreacteds complete
Alkenyl, these groups are the latencies for producing non-specific adsorption, it is necessary to by its inerting.The present invention selects mercaptoethanol conduct
Capping reagent, removes unreacted alkenyl.
It is based on significant advantage and effect described above, of the invention：
The 1st, cysteine is inserted into the C-terminal of recombinant protein A, the sulfydryl that introducing can be with alkenyl characteristic reaction realizes that albumin A exists
Specific site on agarose gel microsphere is fixed.
2nd, from reagent of the allyl glycidyl ether as activated sepharose microballoon, prepare with alkenyl
Activated sepharose microballoon.
3rd, using the sulfydryl that C-terminal is only located on recombinant protein A, it is micro- that albumin A is fixed on Ago-Gel by realization in order
On ball, the Fc lands of albumin A are fully exposed, the absorption property of material is improved.
Advantage above shows as improving clinical therapeutic efficacy, external plasma adsorption test data table on clinical treatment
It is bright, protein A immunoadsorption material of the invention human immunity ball antibody particularly IgG adsorption capacity is reached 60mg/mL with
On, the protein A immunoadsorption material significantly larger than clinically used at present.
Embodiment
Embodiments of the invention described below.It should be understood that embodiments of the invention are used to illustrate rather than
Limitation of the present invention.The improvement carried out according to the essence of the present invention belongs to the scope of protection of present invention.
Embodiment one：The preparation of recombinant protein A with cysteine
The coded sequence of the recombinant protein A with cysteine is obtained by artificial synthesized method, the sequence encodes albumin A
Sequence（SEQ.ID.NO:1）Middle B structure domain is connected, and forms the concatermer in 5 B structure domains, the limitation held positioned at sequence 5 '
Property restriction endonuclease NdeI recognition sequence；The codon tgt of one encoding aminothiopropionic acid and the codon of 6 histidines
Caccaccaccaccaccac is located at the recognition sequence that sequence 3 ' holds XhoI.
SEQ.ID.NO.1
CATATGGTAGACaacaaattcaacaaagaacaacaaaatgctttctatgaaatcttacatttacctaacttaa
acgaagaacaacgcaatggtttcatccaaagcctGaaagatgacccaagccaaagcgctaaccttttagcagaagct
aaaaagctaaatgatgcGcaagcaccaaaaGTAGACaacaaattcaacaaagaacaacaaaatgctttctatgaaat
cttacatttacctaacttaaacgaagaacaacgcaatggtttcatccaaagcctGaaagatgacccaagccaaagcg
ctaaccttttagcagaagctaaaaagctaaatgatgcGcaagcaccaaaaGTAGACaacaaattcaacaaagaacaa
caaaatgctttctatgaaatcttacatttacctaacttaaacgaagaacaacgcaatggtttcatccaaagcctGaa
agatgacccaagccaaagcgctaaccttttagcagaagctaaaaagctaaatgatgcGcaagcaccaaaaGTAGACa
acaaattcaacaaagaacaacaaaatgctttctatgaaatcttacatttacctaacttaaacgaagaacaacgcaat
ggtttcatccaaagcctGaaagatgacccaagccaaagcgctaaccttttagcagaagctaaaaagctaaatgatgc
GcaagcaccaaaaGTAGACaacaaattcaacaaagaacaacaaaatgctttctatgaaatcttacatttacctaact
taaacgaagaacaacgcaatggtttcatccaaagcctGaaagatgacccaagccaaagcgctaaccttttagcagaa
gctaaaaagctaaatgatgcGcaagcaccaaaa GTAGACtgtCTCGAG
By NdeI and XhoI digestions and the effect of T4 DNA ligases, above-mentioned sequence is connected to large intestine bar disclosed in this area
Bacterium expresses bacterial strain pET-22b, and is transformed into E. coli expression strains BL21 (DE3), obtains recombinant protein A expression bacterial strain.
The bacterial strain is cultivated to OD in the fermentation tank of the LB culture mediums equipped with 20L600=1.2, add isopropyl-β-D- sulphur
For galactopyranoside（IPTG）To final concentration of 1mM, continue after fermenting 6 hours, thalline is collected by centrifugation（6000rpm, 20 points
Clock）About 800 grams altogether.The thalline obtained as stated above is mixed in 2 liters of PBS, sonicated cells（Ultrasonic time
5s, interval time 5s, ultrasonic number of times 99 times, power 300W）After be collected by centrifugation（8000rpm, 20 minutes）Supernatant component.Supernatant
Heat half an hour, stand in boiling water bath, centrifugation（8000rpm, 20 minutes）Precipitation is removed, supernatant is affine through chelate cobalt ions
Chromatographic purifying, equilibrium liquid is first to rinse uncombined composition with equilibrium liquid after PBS, loading, then is with containing imidazole concentration
250mM PBS rinses and collects eluent, by obtained Protein A solution in 1M mercaptoethanols (Tris buffer solutions,
PH=8), in reaction 2 hours under 40 DEG C of water bath conditions.Protein liquid is fully dialysed in 0.01% PBS cushioning liquid afterwards, dialysis
Liquid is after 0.45 μm of filtering membrane filtration, and freeze-drying obtains about 4 grams of the albumin A with cysteine.
Embodiment two：The preparation of immune absorption material
The activation of Ago-Gel
60 milliliters of 2.5 M containing 0.2% sodium borohydride are added in the 30 milliliters of Ago-Gel drained Sepharose 6FF
Sodium hydrate aqueous solution is mixed, and is shaken in shaking table（50-200rpm）Reaction 2 hours, reaction temperature is 40 DEG C, is then added
30ml allyl glycidyl ethers continue to react 6 hours.After completion of the reaction, fully rinsed, drained with distilled water, obtain carrying alkene
The Ago-Gel of base.
The fixation of albumin A
Albumin A in 210 milligrams of embodiments one is dissolved in 30 milliliters of 0.1mol/mL phosphate buffer solutions（pH=7.4）In, then
In the Ago-Gel for being added to 30 milliliters of above-mentioned activation（The corresponding albumin A quality 7mg of i.e. every milliliter Ago-Gel）, in purple
Under outer optical wavelength 305nm irradiations, rocked at room temperature（50-200rpm）Reaction about 10 hours, reclaims unreacted Protein A solution, inhales
Attached dose is repeatedly washed with water, is drained.30 milliliters of PBS solutions of mercaptoethanol containing 0.2M are added, are shaken at room temperature（50-200rpm）Instead
Answer 4 hours, after the multiple washes clean of distilled water, drain, be fixed the sorbing material of albumin A.Egg is detected with biuret method
White A fixed amount is 20mg/mL.
Embodiment three：The preparation of sorbing material
The activation of Ago-Gel
Operated with embodiment two with method, obtain the Ago-Gel with alkenyl.
The fixation of albumin A
Albumin A in 180 milligrams of embodiments one is dissolved in 30 milliliters of 0.1mol/mL phosphate buffer solutions（pH=7.4）In, then
In the Ago-Gel for being added to 30 milliliters of above-mentioned activation（The corresponding albumin A quality 6mg of i.e. every milliliter Ago-Gel）, in purple
Under outer optical wavelength 305nm irradiations, rocked at room temperature（50-200rpm）Reaction about 16 hours, reclaims unreacted Protein A solution, inhales
Attached dose is repeatedly washed with water, is drained.30 milliliters of PBS solutions of mercaptoethanol containing 0.2M are added, are shaken at room temperature（50-200rpm）Instead
Answer 4 hours, after the multiple washes clean of distilled water, drain, be fixed the sorbing material of albumin A.Egg is detected with biuret method
White A fixed amount is 25mg/mL.
Example IV：The IgG adsorption capacities of the sorbing material of synthesis are evaluated
1 milliliter of sorbing material in collection embodiment three and example IV, adds human normal plasma 10mL, is shaken at 37 DEG C respectively
Swing 2 hours and carry out adsorption reaction.The suspension is separated 5 minutes with 5000rpm centrifugation, with immune turbidimetry for Determination
IgG concentration in clear liquid.
As check experiment, 1 milliliter of agarose gel microsphere is replaced into sorbing material, with above method same treatment, surveyed
Determine the IgG concentration in solution.
IgG adsorbance is calculated by following formula, as a result as shown in table 1.
IgG adsorbances=(Cr-Ct) × 10 (mg/mL)
Cr：IgG concentration in check experiment solution
Ct：IgG concentration in adsorption test supernatant
Table 1
| Sorbing material | IgG adsorbances（mg/mL） |
| Embodiment two | 68 |
| Embodiment three | 62 |
Embodiment five：The animal experiment of perfusion of extracorporeal circulation column
By 25 milliliters of protein A immunoadsorption material in embodiment two, it is put into beaker and is soaked 1 hour with physiological saline, is loaded on
In a piece 26 × 50mm of φ adsorption column, adsorbent volume is 25mL, and pillar is fully rinsed with physiological saline.With zoopery pig
（Body weight is about 20kg）For subjects, after pig is anaesthetized, extracorporal circulatory system is set up.Start A pumps, from the femoral artery of pig with
60ml/min speed draws blood, and blood plasma is isolated by plasma separator, drives and is entered with 25ml/min speed through B pumps
Adsorption column carries out IgG absorption, mixes and is together pumped into the femoral artery of pig with haemocyte by the blood plasma of adsorption column.Absorption 1 hour
Afterwards, adsorption test is completed.Experimental animal pig after immuno absorbence on the day of recover normal physiological activity, experiment one week after do not find it is different
Normal symptom.
Extract respectively before experiment and the blood plasma of pig carries out the test of IgG concentration after experiment, calculate IgG rate of descent.Under IgG
Drop rate is calculated according to below equation, as a result as shown in table 2.
Rate of descent=（Cb-Ca）/ Cb × 100%
Cb is the IgG concentration of pig before experiment
Ca is the IgG concentration of pig after experiment
Embodiment six：The animal experiment of perfusion of extracorporeal circulation column
In addition to " protein A immunoadsorption material in embodiment two " is changed into " protein A immunoadsorption material in embodiment three ",
It is other to be operated with embodiment five with method.
Table 2
| Extracorporeal circulation column | IgG rates of descent |
| Embodiment five | 46.8% |
| Embodiment six | 42.3% |
Industrial applicability
From above-described embodiment, the invention provides a kind of new blood purification Protein A immunoadsorption agent, it can pacify
Entirely, IgG antibody reliably, optionally in efficient absorption human plasma.In addition, being followed using the external of above-mentioned sorbing material is filled
Annulated column, can optionally remove the IgG antibody in blood plasma by disposable perfusion.
Sequence table（SEQUENCE LISTING）
<110>Chongqing Xi'erkang Blood Purification Equipment Research ＆ Development Co., Ltd.
<120>A kind of immune absorption material for pinpointing fixed protein A and preparation method thereof
<160> 1
<210> 1
<211> 891
<212> DNA
<213>Artificial sequence
<400> 1
catatggtag acaacaaatt caacaaagaa caacaaaatg ctttctatga aatcttacat 60
ttacctaact taaacgaaga acaacgcaat ggtttcatcc aaagcctgaa agatgaccca 120
agccaaagcg ctaacctttt agcagaagct aaaaagctaa atgatgcgca agcaccaaaa 180
gtagacaaca aattcaacaa agaacaacaa aatgctttct atgaaatctt acatttacct 240
aacttaaacg aagaacaacg caatggtttc atccaaagcc tgaaagatga cccaagccaa 300
agcgctaacc ttttagcaga agctaaaaag ctaaatgatg cgcaagcacc aaaagtagac 360
aacaaattca acaaagaaca acaaaatgct ttctatgaaa tcttacattt acctaactta 420
aacgaagaac aacgcaatgg tttcatccaa agcctgaaag atgacccaag ccaaagcgct 480
aaccttttag cagaagctaa aaagctaaat gatgcgcaag caccaaaagt agacaacaaa 540
ttcaacaaag aacaacaaaa tgctttctat gaaatcttac atttacctaa cttaaacgaa 600
gaacaacgca atggtttcat ccaaagcctg aaagatgacc caagccaaag cgctaacctt 660
ttagcagaag ctaaaaagct aaatgatgcg caagcaccaa aagtagacaa caaattcaac 720
aaagaacaac aaaatgcttt ctatgaaatc ttacatttac ctaacttaaa cgaagaacaa 780
cgcaatggtt tcatccaaag cctGaaagat gacccaagcc aaagcgctaa ccttttagca 840
gaagctaaaa agctaaatga tgcgcaagca ccaaaagtag actgtctcga g 891
Claims (9)
1. a kind of recombinant protein A immune absorption material for being used to remove antibody, is that recombinant protein A is covalently bonded into agarose to coagulate
High polymer material on glue microballoon, it has following chemical constitution：
Picture 11
Wherein：
Represent agarose gel microsphere；RSPA represents recombinant protein A；（C）Represent the C-terminal key of recombinant protein A
Close on agarose gel microsphere.
2. the immune absorption material in claim 1, it is characterised in that recombinant protein A is fixed on agarose gel microsphere and made
Standby to form, material absorption amount of antibody reaches more than 60mg/mL.
3. the chelate cobalt ions affinitive layer purification of the recombinant protein A described in claim 1.
4. the gene order of the recombinant protein A described in claim 1 is by albumin A coded sequence（SEQ.ID.NO:1）Middle B knots
Structure domain is connected, the concatermer in 5 B structure domains of formation.
5. the albumin A described in claim 1 is handled with mercaptoethanol after purification, prevent from forming two sulphur between sulfydryl
Key.
6. the fixed amount of the albumin A described in claim 1 reaches more than 20mg/mL.
7. the preparation method of the recombinant protein A immune absorption material in claim 1, it is characterized in that：Using Ago-Gel as load
Body, will Ago-Gel and allyl glycidyl ether react after form the activated sepharose with double bond, then with again
Histone A C-terminal sulfydryl coupling, forms protein A immunoadsorption material, is characterized in particular in：
In 10 ~ 12mol/L alkaline aqueous solution, 4 are reacted in 40 ~ 60 DEG C with agarose gel microsphere for allyl glycidyl ether
~ 8 hours Ago-Gels volume ratio shared in reaction solution is 20% ~ 40%；
After step a reactions terminate, reaction product is filtered, with distilled water flushing to neutrality, the activation agar with double bond is obtained
Sugared gel micro-ball；
Product obtained by step b is added in phosphate buffer solution, and control ph adds recombinant protein A, in purple in the range of 6 ~ 8
Outer optical wavelength 305nm irradiations are lower to react.
8. reaction temperature is 20 ~ 25 DEG C, the reaction time is 10 ~ 16 hours.
9. products therefrom adds mercaptoethanol and carries out end-block reaction in step c, the concentration of capping reagent is 0.1 ~ 0.5mol/
L, pH value is 6 ~ 8, and the reaction time is 4 ~ 6 hours.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611251363.8A CN107051396A (en) | 2016-12-30 | 2016-12-30 | A kind of immune absorption material for pinpointing fixed protein A and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611251363.8A CN107051396A (en) | 2016-12-30 | 2016-12-30 | A kind of immune absorption material for pinpointing fixed protein A and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107051396A true CN107051396A (en) | 2017-08-18 |
Family
ID=59623797
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611251363.8A Pending CN107051396A (en) | 2016-12-30 | 2016-12-30 | A kind of immune absorption material for pinpointing fixed protein A and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107051396A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107670649A (en) * | 2017-10-20 | 2018-02-09 | 云南师范大学 | The active carrier and preparation method and the preparation method of protein A immunoadsorption material of a kind of directional at-tachment albumin A |
| CN110624274A (en) * | 2019-08-27 | 2019-12-31 | 苏州赛分科技有限公司 | Separation medium, preparation method and application thereof |
| CN110681362A (en) * | 2019-09-26 | 2020-01-14 | 浙江大学 | Mixed-mode chromatography medium with carboxyl and indolyl as functional groups |
| CN111233985A (en) * | 2020-02-09 | 2020-06-05 | 北京丹大生物技术有限公司 | Preparation method of novel coronavirus IgA antibody rapid detection test strip |
| CN111551753A (en) * | 2019-10-02 | 2020-08-18 | 华中科技大学同济医学院附属协和医院 | ABO blood group immunoadsorption membrane and preparation method thereof |
| CN116850977A (en) * | 2023-07-28 | 2023-10-10 | 广州康盛生物科技股份有限公司 | Directional coupling immunoadsorbent as well as preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1365853A (en) * | 2001-01-17 | 2002-08-28 | 玉环县卫康医疗器械有限公司 | Affinity adsorption medium and its preparing medium |
| CN101602805A (en) * | 2009-07-08 | 2009-12-16 | 中国科学院过程工程研究所 | Be used for agarose compatible medium of purifying hand foot mouth disease immunoglobulin (Ig) and preparation method thereof |
| CN102614843A (en) * | 2012-04-05 | 2012-08-01 | 云南澳敦生物科技有限公司 | Protein A immunosorbent material and preparation method thereof |
| CN102716719A (en) * | 2012-07-04 | 2012-10-10 | 汪志友 | Heavy metal adsorbent for extracorporeal perfusion method and preparation method of heavy metal absorbent |
-
2016
- 2016-12-30 CN CN201611251363.8A patent/CN107051396A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1365853A (en) * | 2001-01-17 | 2002-08-28 | 玉环县卫康医疗器械有限公司 | Affinity adsorption medium and its preparing medium |
| CN101602805A (en) * | 2009-07-08 | 2009-12-16 | 中国科学院过程工程研究所 | Be used for agarose compatible medium of purifying hand foot mouth disease immunoglobulin (Ig) and preparation method thereof |
| CN102614843A (en) * | 2012-04-05 | 2012-08-01 | 云南澳敦生物科技有限公司 | Protein A immunosorbent material and preparation method thereof |
| CN102716719A (en) * | 2012-07-04 | 2012-10-10 | 汪志友 | Heavy metal adsorbent for extracorporeal perfusion method and preparation method of heavy metal absorbent |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107670649A (en) * | 2017-10-20 | 2018-02-09 | 云南师范大学 | The active carrier and preparation method and the preparation method of protein A immunoadsorption material of a kind of directional at-tachment albumin A |
| CN107670649B (en) * | 2017-10-20 | 2020-04-07 | 云南师范大学 | Active carrier for directionally fixing protein A, preparation method and preparation method of protein A immunoadsorption material |
| CN110624274A (en) * | 2019-08-27 | 2019-12-31 | 苏州赛分科技有限公司 | Separation medium, preparation method and application thereof |
| CN110624274B (en) * | 2019-08-27 | 2021-04-13 | 苏州赛分科技有限公司 | Separation medium, preparation method and application thereof |
| CN110681362A (en) * | 2019-09-26 | 2020-01-14 | 浙江大学 | Mixed-mode chromatography medium with carboxyl and indolyl as functional groups |
| CN111551753A (en) * | 2019-10-02 | 2020-08-18 | 华中科技大学同济医学院附属协和医院 | ABO blood group immunoadsorption membrane and preparation method thereof |
| CN111233985A (en) * | 2020-02-09 | 2020-06-05 | 北京丹大生物技术有限公司 | Preparation method of novel coronavirus IgA antibody rapid detection test strip |
| CN116850977A (en) * | 2023-07-28 | 2023-10-10 | 广州康盛生物科技股份有限公司 | Directional coupling immunoadsorbent as well as preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107051396A (en) | A kind of immune absorption material for pinpointing fixed protein A and preparation method thereof | |
| EP0237659B1 (en) | Process for the production of protein a-silica immunoadsorbent | |
| CN107670649B (en) | Active carrier for directionally fixing protein A, preparation method and preparation method of protein A immunoadsorption material | |
| CN102614843B (en) | Protein A immunoadsorption material and preparation method thereof | |
| CN102233128B (en) | Use of p75neurotrophin receptor-extracellular domain (p75NTR-ECD) in medicine for preventing and treating Alzheimer disease | |
| JP5774607B2 (en) | Removal of toxic factors by in vitro treatment | |
| CN101190409B (en) | Blood purifying protein A immunoadsorption material and synthesizing method thereof | |
| CN111057153B (en) | Immunoglobulin binding protein and preparation method and application thereof | |
| CN107486176A (en) | A kind of sorbing material for blood purification and preparation method thereof | |
| CN107754764B (en) | High-load protein A immunoadsorption material and preparation method thereof | |
| CN101185878B (en) | Protein A immunoadsorption material for eliminating pathogenic antibody and its complexes, and synthesizing method and application thereof | |
| CN101069751B (en) | Protein A immuo adsorption material and preparing method | |
| CN108752468A (en) | Wound spray | |
| CN101478998B (en) | substrate for biological fluid treatment | |
| WO1990004416A1 (en) | Specific removal of ldl from blood | |
| EP1570272A2 (en) | Identification of agonistic autoantibodies | |
| CN103111270A (en) | Adsorbing material of hepatitis B antigen protein and preparation method of material | |
| CN1100568C (en) | Process for synthesizing protein immunoadsorbent medium for removing pathogenic antibody and its compounds from plasm | |
| JPH02231078A (en) | Superoxide dismutase modified with hyaluronic acid | |
| CN101525598B (en) | Method for purifying human urinary trypsin inhibitor | |
| CN1293245A (en) | Process for separating worm kinase | |
| US4608253A (en) | Process for removing immune complex in blood by use of the immobilized pepsin | |
| WO2008142694A2 (en) | Peptides for the treatment of systemic lupus erythematosus and methods of treating systemic lupus erythematosus | |
| CN108816208B (en) | High-load protein A immunoadsorption material and preparation method thereof | |
| CN117024565A (en) | Polypeptide modified body and application thereof in preparation of anti-escherichia coli infection drugs |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | 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: 20170818 |
|
| WD01 | Invention patent application deemed withdrawn after publication |