CN1958599B - Method for enriching and purifying glycosylation protein - Google Patents
Method for enriching and purifying glycosylation protein Download PDFInfo
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- CN1958599B CN1958599B CN2005100962708A CN200510096270A CN1958599B CN 1958599 B CN1958599 B CN 1958599B CN 2005100962708 A CN2005100962708 A CN 2005100962708A CN 200510096270 A CN200510096270 A CN 200510096270A CN 1958599 B CN1958599 B CN 1958599B
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Abstract
This invention discloses a method for enriching and purifying glycosylated protein. The immobilization of agglutinin comprises: adding epoxy ethyl derived magnetic particles into a centrifugal tube, performing immobilization reaction, washing, sealing, washing repeatedly to immobilize agglutinin onto epoxy ethyl derived magnetic particles, adding buffer and storing. The separation and purification of glycosylated protein comprises: immobilizing prepared agglutinin onto epoxy ethyl derived magnetic particles, adding standard glycosylated protein for adequate coupling, washing repeatedly, eluting to obtain enriched and purified glycosylated protein, loading into a dialyzing bag, and desalting. This invention solves the problems of complex enriching and separation process, low recovery rate and high cost faced by background technology. This invention has such advantages as simple process, high recovery rate, low affinity chromatographic column cost, and convenient usage.
Description
Technical field
What the present invention relates to is a kind of lectin to be connected on the magnetic particle after the modification, utilize thisly to be fixed on lectin on the magnetic particle specifically in conjunction with glycosylated protein, thereby to the method for its separation, enrichment and purifying.
Background technology
Lectin (Lectins) is the protein or the glycoprotein of a class non-antibody, non-covalent combination that can be single-minded with carbohydrate, and have aggegation cell and the effect that precipitates glycan and compounding sugar.Therefore its structure contains carbohydrate differential threshold (CRD), and can single-minded the combination be taken place with carbohydrate, and the structure or the conformation of different configuration, glycosidic link type and the oligonucleotide chain of residue had specificity.As to accompany ConA (Con A) be a kind of lectin from sword bean, can be single-minded in conjunction with α-seminose sugar chain inside or non-reducing end (Man).And companion's ConA can be connected on the solid support by appropriate chemical methods, be rich in mannose residue in the high mannose type glycoprotein that N-connects, therefore can utilize the companion's ConA purification high mannose type glycoprotein from egg white mixture on the solid support.At present, the existing abroad commercial affinity column that is used to separate some specific structure glycosylated proteins.
Mainly there is following shortcoming in existing glycosylated protein separation and concentration technology:
1. be the affinity column that is used to separate some specific structure glycosylated proteins owing to having commercial.Affinitive layer purification must be at a certain separate object, prepare single-minded aglucon and seek the stable condition of chromatography, this will seek a marriage alliance and the special post special use of chromatography column, i.e. pillar class glycoprotein of can only purifying, therefore the range of application of affinity chromatography has been subjected to certain restriction, and method is comparatively complicated.
2. the affinity column of separation and concentration glycosylated protein is lower to the rate of recovery of glycosylated protein.
3. the affinity column of separation and concentration glycosylated protein costs an arm and a leg.
Summary of the invention
The object of the present invention is to provide a kind of method based on micro-nano magnetic technique separation, enrichment and purifying glycosylation protein, it has solved in the background technology and with affinity column glycosylated protein method for separating and concentrating complexity, the rate of recovery has been hanged down and expensive technical problem.
Technical solution of the present invention is:
The method of a kind of enrichment and purifying glycosylation protein is characterized in that: this method may further comprise the steps
1) immobilization of lectin
(1) epoxy ethyl derivatize magnetic grain is prefabricated: the magnetic grain of epoxy ethyl derivatize is added in the centrifuge tube, be designated as centrifuge tube No. 1; Add coupling buffer again, shake up, on magnetic separator, separate, abandon supernatant liquor;
(2) immobilized reactant: get lectin solution, be added in No. 1 centrifuge tube, shake up; Place Air oscillator, 35～40 ℃ of reactions Celsius 40～90 minutes; Reaction finishes, and separates, abandons supernatant liquor on magnetic separator; Then, the lectin immobilization in No. 1 centrifuge tube is to epoxy ethyl derivatize magnetic grain;
(3) clean: in No. 1 centrifuge tube, add cleaning buffer solution, shake up, on magnetic separator, separate, abandon supernatant liquor;
(4) sealing: in No. 1 centrifuge tube, add confining liquid, shake up, place Air oscillator, 35～40 ℃ of reactions Celsius 1～2 hour; Reaction finishes, and separates, abandons supernatant liquor on magnetic separator;
(5) clean: add cleaning buffer solution in No. 1 centrifuge tube, mixing separates, abandons supernatant liquor on magnetic separator; Repeated washing 3 times;
(6) preserve: add and preserve damping fluid, in 2～6 ℃ of preservations Celsius;
2) separation and purification of glycosylated protein
(1) prefabricated: as to get the lectin immobilization for preparing and arrive epoxy ethyl derivatize magnetic grain, add No. 4 centrifuge tube, on magnetic separator, separate, abandon supernatant liquor;
(2) combination: in No. 4 centrifuge tubes, add binding buffer liquid, add the standard glycosylated protein again, and then add ultrapure water, place on the shaking table, at room temperature shook 4～6 hours; With No. 4 centrifuge tubes, on magnetic separator, separate, abandon supernatant liquor;
(3) clean: in No. 4 centrifuge tubes, add cleaning buffer solution 1, place on the shaking table, shook 1～3 minute, on magnetic separator, separate, abandon supernatant liquor; Repeated washing once again;
(4) clean: in No. 4 centrifuge tubes, add cleaning buffer solution 2, cleaned 1～3 minute, on magnetic separator, separate, abandon supernatant liquor; Repeated washing secondary again;
(5) wash-out: add cleaning buffer solution 3 in No. 4 centrifuge tubes, 20～30 ℃ Celsius were cleaned 20～40 minutes, separated on magnetic separator, got in No. 5 centrifuge tubes of supernatant liquor immigration, promptly got the glycosylated protein of enriching and purifying;
(6) desalination: the glycosylated protein of enriching and purifying is packed in the dialysis tubing, and water changes water in 2～6 ℃ of dialysis Celsius after 1.5～2.5 hours, repeats to dialyse three times, sloughs cleaning buffer solution 3.
In the immobilization of above-mentioned lectin, described coupling buffer is to adopt the NaHCO of pH8.3 0.1M
30.5M the NaCl damping fluid be advisable; Also can employing or damping fluids such as phosphoric acid salt, carbonate or acetate.
In the immobilization of above-mentioned lectin, described confining liquid is the skim-milk of adding 1mg/ml or the damping fluids such as phosphoric acid salt, carbonate or acetate of bovine serum albumin.
In the immobilization of above-mentioned lectin, described cleaning buffer solution, preserve damping fluid, also can adopt or carbonate buffer solution all to adopt 0.5% the tween 20 1 * phosphoric acid salt that contains to be advisable, or acetate buffer etc.
In the separation and purification of above-mentioned glycosylated protein, described binding buffer liquid is to adopt 1mM CaCl
2, 1mMMnCl
2, 10mM Tris-HCl, pH 7.0 damping fluid be advisable, also can adopt damping fluids such as phosphoric acid salt, carbonate or acetate; Described cleaning buffer solution 1 is to adopt 1mM CaCl
2, 1mM MnCl
2, 10mM Tris-HCl, pH 7.0 damping fluid be advisable, also can adopt damping fluids such as phosphoric acid salt, carbonate or acetate; Described cleaning buffer solution 2 is to adopt 1mM CaCl
2, 1mM MnCl
2, 10mM Tris-HCl, pH 7.0 damping fluid be advisable, also can adopt damping fluids such as phosphoric acid salt, carbonate or acetate.
In the separation and purification of above-mentioned glycosylated protein, cleaning buffer solution 3 is to adopt 1mM CaCl
2, 1mM MnCl
2, 1mM KCl, 100mM Alpha-Methyl seminose, 10mM Tris-HCl, pH 7.0 damping fluid be advisable, also can adopt damping fluids such as phosphoric acid salt, carbonate or acetate; Described regeneration damping fluid 1 is to adopt 1mM CaCl
2, 1mM MnCl
2, 10mM Tris-HCl, pH 4.5 damping fluid be advisable, also can adopt damping fluids such as phosphoric acid salt, carbonate or acetate; Described regeneration damping fluid 2 is to adopt 1mM CaCl
2, 1mM MnCl
2, 10mMTris-HCl, pH 8.0 damping fluid be advisable, also can adopt damping fluids such as phosphoric acid salt, carbonate or acetate.
In the immobilization of above-mentioned lectin, the temperature of reaction on Air oscillator is advisable with 37 ℃ Celsius, and the reaction times was advisable with 60 minutes; Described lectin immobilization is advisable with 4 ℃ Celsius to the storage temperature of epoxy ethyl derivatize magnetic grain; In the separation and purification of described glycosylated protein, the dialysis temperature of the glycosylated protein of enriching and purifying is advisable with 4 ℃ Celsius, and the time of each dialysis was advisable with 1 hour.
Above-mentioned glycosylated protein is after separation and purification, and the lectin immobilization is renewable to the lectin on the epoxy ethyl derivatize magnetic grain, and regeneration step comprises:
1) regeneration: add regeneration damping fluid 1 in No. 4 centrifuge tubes, 20～30 ℃ Celsius were cleaned 20～40 minutes, separated, abandon supernatant liquor on magnetic separator; Repeat regeneration once;
2) preserve: in No. 4 centrifuge tubes, add and preserve damping fluid, in 2～6 ℃ of preservations Celsius.
Above-mentioned epoxy ethyl derivatize magnetic grain is after prefabricated, immobilized reactant, cleaning, and the lectin immobilization is to epoxy ethyl derivatize magnetic grain, and this lectin immobilization can detect to the immobilization efficiency on the epoxy ethyl derivatize magnetic grain, detects step and comprises
1) with in the immobilized reactant: the supernatant liquor liquid that No. 1 centrifuge tube is abandoned on magnetic separator moves in No. 2 centrifuge tubes;
2) in the cleaning behind immobilized reactant: the supernatant liquor that No. 1 centrifuge tube is abandoned on magnetic separator is moved in No. 3 centrifuge tubes;
3) detection of immobilization efficiency: is blank with the coupling buffer, gets the solution in lectin solution and No. 2 centrifuge tubes, both are remembered at the absorbance at 280nm place make OD
1And OD
2With the cleaning buffer solution is blank, and solution is made OD in the absorbance note at 280nm place in No. 3 centrifuge tubes
3, then coupling efficiency is
Coupling efficiency %=(OD
1-OD
2-OD
3)/OD
1
The present invention has the following advantages:
1. the method for separation, enrichment and purifying glycosylation protein is easy.
2. the rate of recovery is higher.
3. the affinity column cost of separation and concentration glycosylated protein is low.
4. easy-to-use.Time spent can connect at any time, is not subjected to the influence of lectin kind.
Description of drawings
Fig. 1 is a principle schematic of the present invention;
Fig. 2 is the SDS-PAGE electrophoretic effects synoptic diagram of the glycosylated protein of enriching and purifying of the present invention.
The explanation of accompanying drawing drawing: 1-swimming lane 1 is albumen Mark; 2-swimming lane 2 is the albumen swimming lanes that are not connected on the magnetic particle; 3-swimming lane 3 is to wash next albumen swimming lane by washing lotion 1; 4-swimming lane 4 is to wash next albumen swimming lane by washing lotion 2; 5-swimming lane 5 is to wash next albumen swimming lane by washing lotion 3.
Embodiment
The present invention is connected to the nano level magnetic particle of functionalization with lectin, on the nano level magnetic particle as epoxy ethyl, amino derivatization, is used for the separation and concentration glycosylated protein.Glycosylated protein is to form after protein is connected with monose chain covalency, is mainly used in cell recognition, and physiological function is mainly used in immune cell recognition etc.
The performing step of the embodiment of the invention is as follows:
1) immobilization of lectin
(1) epoxy ethyl derivatize magnetic grain is prefabricated: after the magnetic grain of epoxy ethyl derivatize is shaken up, pipette 200 μ l in centrifuge tube with pipettor, be designated as centrifuge tube No. 1.In No. 1 centrifuge tube, add 400 μ l coupling buffers,, on magnetic separator, separated 3 minutes, keep No. 1 centrifuge tube on magnetic separator, pipette supernatant liquor with pipettor and discard with hand even.
(2) immobilized reactant: get lectin solution, as companion's ConA (Con A) solution 300 μ L, about 150 μ g are added in No. 1 centrifuge tube, shake up, and are placed in the Air oscillator, and reaction is 1 hour under 37 ℃ of conditions.Reaction finishes, and magnetic resolution 3 minutes keeps No. 1 centrifuge tube on magnetic separator, pipettes supernatant liquor in No. 2 centrifuge tubes with pipettor.Then, the companion's ConA immobilization in No. 1 centrifuge tube is to epoxy ethyl derivatize magnetic grain.
(3) clean: in No. 1 centrifuge tube, add 400 μ l cleaning buffer solutions, spare, place on the magnetic separator, separated 3 minutes, keep No. 1 centrifuge tube on magnetic separator, pipette supernatant liquor in No. 3 centrifuge tubes with pipettor with hand.
(4) detection of immobilization efficiency: is blank with the coupling buffer, gets the solution in 800 μ l companion's ConA solution and No. 2 centrifuge tubes, both remember at the absorbance at 280nm place and make OD
1And OD
2With the cleaning buffer solution is blank, and solution is made OD in the absorbance note at 280nm place in No. 3 centrifuge tubes
3, then coupling efficiency is
Coupling efficiency %=(OD
1-OD
2-OD
3)/OD
1
(5) sealing: No. 1 centrifuge tube is placed on the magnetic separator, separated 3 minutes, keep No. 1 centrifuge tube on magnetic separator, pipette supernatant with pipettor and discard.Add confining liquid 800 μ l in No. 1 centrifuge tube, the jog mixing places Air oscillator, and reaction is 2 hours under 37 ℃ of conditions.Reaction finishes, and magnetic resolution 3 minutes keeps No. 1 centrifuge tube on magnetic separator, pipettes supernatant liquor with pipettor and discards.
(6) clean: in No. 1 centrifuge tube, add the cleaning buffer solution of 800 μ l, use the pipettor mixing, placed on the magnetic separator separation 3 minutes, keep No. 1 centrifuge tube on magnetic separator, pipette supernatant liquor with pipettor and discard, repeat this step 3 time.
(7) preserve: add the preservation damping fluid of 1ml in the epoxy ethyl derivatize magnetic grain in the immobilization of companion's ConA, in 4 ℃ of preservations.
2) separation and purification of glycosylated protein
Glycoprotein in the present embodiment is high mannose type albumen ribonuclease B, and ribonuclease B is the proteolytic enzyme that a kind of seminose is modified, its can with companion's ConA specific combination.
(1) companion's ConA immobilization prefabricated to epoxy ethyl derivatize magnetic grain: get the companion's ConA immobilization for preparing and arrive epoxy ethyl derivatize magnetic grain, add No. 4 centrifuge tube, on magnetic separator, separated 3 minutes, keep No. 4 centrifuge tubes on magnetic separator, draw supernatant liquor with pipettor and discard.
(2) combination: in No. 4 centrifuge tubes, add the binding buffer liquid of 200 μ l, as 1mM CaCl
2, 1mMMnCl
2, the damping fluid of 10mM Tris-HCl pH7.0; The standard glycosylated protein that adds 2mg/ml again, as: ribonuclease B 300 μ l; Add 300 μ l ultrapure waters again, place on the shaking table, at room temperature shook 4～6 hours, make it abundant coupling.Take out No. 4 centrifuge tube, shake up to be placed on the magnetic separator and separated 3 minutes, pipette supernatant liquor in No. 5 centrifuge tubes.
(3) clean: in No. 4 centrifuge tubes, add the cleaning buffer solution 1 of 400 μ l, as 1mMCaCl
2, 1mMMnCl
2, 10mM Tris-HCl pH7.0
4Damping fluid, on shaking table, shook 2 minutes, place on the magnetic separator and to separate 3 minutes, draw supernatant liquor and place centrifuge tube No. 6; Repeated washing once.
(4) clean: in No. 4 centrifuge tubes, add the cleaning buffer solution 2 of 400 μ l, as 1mMCaCl
2, 1mMMnCl
2, 10mM Tris-HCl pH7.0 damping fluid, cleaned 2 minutes, place on the magnetic separator and to separate 3 minutes, draw supernatant and place centrifuge tube No. 7; Repeated washing is twice again.Cleaning buffer solution 1,2 is used to remove the albumen of non-specific binding.
(5) wash-out: in No. 4 centrifuge tubes, add 800 μ l and wash down damping fluid liquid 3, as 1mM CaCl
2, 1mMMnCl
2, 1mMKCl, 100mM Alpha-Methyl seminose (a-methyl mannose), 10mMTris-HCl pH7.0; Cleaned 30 minutes in 25 ℃.Place on the magnetic separator and to separate 3 minutes, draw supernatant in No. 8 centrifuge tubes, be the high mannose type albumen of enriching and purifying.
(6) desalination: the high mannose type albumen of enriching and purifying is packed in the dialysis tubing, change water, triplicate in 4 ℃ after dialysing 2 hours with 1L water.
(7) detect: with the albumen of SDS-PAGE electrophoresis detection enriching and purifying, referring to Fig. 2.The Alpha-Methyl seminose is arranged in the cleaning buffer solution 3, and its specific combination ability with companion's ConA is stronger with companion's ConA specific combination ability than ribonuclease B.At this competitive combination has taken place, and ribonuclease B is washed, so the band of tangible ribonuclease B has appearred in swimming lane 5.
(8) regeneration: in No. 4 centrifuge tubes, add the regeneration damping fluid 1 of 800 μ l, as 1mM CaCl2,1mMMnCl2,10mM Tris-HCl pH 4.5 cleaned 30 minutes in 25 ℃.Place on the magnetic separator and separated 3 minutes, draw supernatant liquor, discard.
(9) regeneration: in No. 4 centrifuge tubes, add the regeneration damping fluid 2 of 800 μ l, as 1mM CaCl2,1mMMnC12,10mM Tris-HCl pH cleans about 30min for 8.0,25 ℃.Put on the magnetic separator and separated 3 minutes, draw supernatant liquor, discard.
(10) preserve: in No. 4 centrifuge tubes, add the preservation damping fluid of 500 μ l, as 1mM CaCl2,1mMMnCl2,0.02%NaN3,10mM Tris-HCl pH 6.0 is in 4 ℃ of preservations.
This companion's ConA immobilization that is fixed on can repeatedly be regenerated to the lectin on the epoxy ethyl derivatize magnetic grain, reuses, and the number of times of enriching and purifying glycoprotein is at least 10 times.
Glycoprotein in the present embodiment is high mannose type albumen ribonuclease B, ribonuclease B is the proteolytic enzyme that a kind of seminose is modified, because it has and companion's ConA (Con A) specific combination ability, so the amount that is washed out by cleaning buffer solution 1,2 is few, and seminose is arranged in the cleaning buffer solution 3, can with the competitive combination of ribonuclease B, and Yeast Nucleic Acid B is washed, so the band of swimming lane 5 is more obvious.Referring to Fig. 2.
Claims (1)
1. the method for enrichment and purifying glycosylation protein, it is characterized in that: this method may further comprise the steps
1) immobilization of lectin
(1) epoxy ethyl derivatize magnetic grain is prefabricated: the magnetic grain of epoxy ethyl derivatize is added in the centrifuge tube, be designated as centrifuge tube No. 1; Add coupling buffer again, shake up, on magnetic separator, separate, abandon supernatant liquor; Described coupling buffer is the NaHCO of pH8.30.1M
30.5M the NaCl damping fluid;
(2) immobilized reactant: get lectin solution, be added in No. 1 centrifuge tube, shake up; Place Air oscillator, 35～40 ℃ of reactions Celsius 40～90 minutes; Reaction finishes, and separates, abandons supernatant liquor on magnetic separator; Then, the lectin immobilization in No. 1 centrifuge tube is to epoxy ethyl derivatize magnetic grain;
(3) clean: in No. 1 centrifuge tube, add cleaning buffer solution, shake up, on magnetic separator, separate, abandon supernatant liquor;
(4) sealing: in No. 1 centrifuge tube, add confining liquid, shake up, place Air oscillator, 35～40 ℃ of reactions Celsius 1～2 hour; Reaction finishes, and separates, abandons supernatant liquor on magnetic separator; Described confining liquid is the skim-milk of adding 1mg/ml or phosphoric acid salt, carbonate or the acetate buffer of bovine serum albumin;
(5) clean: add cleaning buffer solution in No. 1 centrifuge tube, mixing separates, abandons supernatant liquor on magnetic separator; Repeated washing 3 times; Described cleaning buffer solution be 0.5% contain tween 20 1 * phosphoric acid salt or carbonate buffer solution;
(6) preserve: add and preserve damping fluid, in 2～6 ℃ of preservations Celsius; Described preservation damping fluid be 0.5% contain tween 20 1 * phosphoric acid salt or carbonate buffer solution;
2) separation and purification of glycosylated protein
(1) prefabricated: as to get the lectin immobilization for preparing and arrive epoxy ethyl derivatize magnetic grain, add No. 4 centrifuge tube, on magnetic separator, separate, abandon supernatant liquor;
(2) combination: in No. 4 centrifuge tubes, add binding buffer liquid, add the standard glycosylated protein again, and then add ultrapure water, place on the shaking table, at room temperature shook 4～6 hours; With No. 4 centrifuge tubes, on magnetic separator, separate, abandon supernatant liquor; Described binding buffer liquid is 1mM CaCl
2, 1mM MnCl
2, 10mMTris-HCl pH7.0 damping fluid;
(3) clean: in No. 4 centrifuge tubes, add cleaning buffer solution 1, place on the shaking table, shook 1～3 minute, on magnetic separator, separate, abandon supernatant liquor; Repeated washing once again; Described cleaning buffer solution 1 is 1mM CaCl
2, 1mM MnCl
2, 10mM Tris-HCl pH 7.0 damping fluid;
(4) clean: in No. 4 centrifuge tubes, add cleaning buffer solution 2, cleaned 1～3 minute, on magnetic separator, separate, abandon supernatant liquor; Repeated washing secondary again; Described cleaning buffer solution 2 is 1mM CaCl
2, 1mM MnCl
2, 10mM Tris-HCl pH 7.0 damping fluid;
(5) wash-out: add cleaning buffer solution 3 in No. 4 centrifuge tubes, 20～30 ℃ Celsius were cleaned 20～40 minutes, separated on magnetic separator, got in No. 5 centrifuge tubes of supernatant liquor immigration, promptly got the glycosylated protein of enriching and purifying; Described cleaning buffer solution 3 is 1mM CaCl
2, 1mMMnCl
2, 1mMKCl, 100mM Alpha-Methyl seminose, 10mM Tris-HCl pH 7.0 damping fluid;
(6) desalination: the glycosylated protein of enriching and purifying is packed in the dialysis tubing, and water changes water in 2～6 ℃ of dialysis Celsius after 1.5～2.5 hours, repeats to dialyse three times, sloughs cleaning buffer solution 3.
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| CN2005100962708A CN1958599B (en) | 2005-11-03 | 2005-11-03 | Method for enriching and purifying glycosylation protein |
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| CN1958599B true CN1958599B (en) | 2010-11-17 |
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| CN102190701B (en) * | 2010-03-05 | 2013-01-23 | 西北大学 | Method for separating and purifying influenza virus hemagglutinin on large scale |
| CN102268064B (en) * | 2010-06-01 | 2015-04-29 | 中国医学科学院基础医学研究所 | Glycopeptide enrichment and separation material and application thereof |
| CN107903301A (en) * | 2017-12-22 | 2018-04-13 | 广州誉嘉生物科技有限公司 | A kind of separation method for glycosylating albumen |
| CN108387435B (en) * | 2018-01-30 | 2020-09-15 | 中国计量大学 | Trace silk fibroin enrichment method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004064608A2 (en) * | 2003-01-17 | 2004-08-05 | Aethlon Medical, Inc. | Method for removal of viruses from blood by lectin affinity hemodialysis |
| WO2005020936A3 (en) * | 2003-02-28 | 2005-06-23 | Antigenics Inc | Use of lectins to promote oligomerization of glycoproteins and antigenic molecules |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004064608A2 (en) * | 2003-01-17 | 2004-08-05 | Aethlon Medical, Inc. | Method for removal of viruses from blood by lectin affinity hemodialysis |
| WO2005020936A3 (en) * | 2003-02-28 | 2005-06-23 | Antigenics Inc | Use of lectins to promote oligomerization of glycoproteins and antigenic molecules |
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Owner name: SHAANXI BAIMEI GENE CO., LTD. Free format text: FORMER NAME: SHAANXI LIFEGEN CO., LTD. |
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Address after: 386 box 710069, Northwestern University, 229 Taibai North Road, Shaanxi, Xi'an Patentee after: Shaanxi Baimei Gene Co., Ltd. Address before: 386 box 710069, Northwestern University, 229 Taibai North Road, Shaanxi, Xi'an Patentee before: Shaanxi Beimei Gene Co., Ltd. |