CN112794425A - Method for improving flocculation capacity of dextran - Google Patents
Method for improving flocculation capacity of dextran Download PDFInfo
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- CN112794425A CN112794425A CN202110164170.3A CN202110164170A CN112794425A CN 112794425 A CN112794425 A CN 112794425A CN 202110164170 A CN202110164170 A CN 202110164170A CN 112794425 A CN112794425 A CN 112794425A
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/52—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities
- C02F1/5263—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities using natural chemical compounds
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/52—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities
- C02F1/54—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities using organic material
- C02F1/56—Macromolecular compounds
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0009—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid alpha-D-Glucans, e.g. polydextrose, alternan, glycogen; (alpha-1,4)(alpha-1,6)-D-Glucans; (alpha-1,3)(alpha-1,4)-D-Glucans, e.g. isolichenan or nigeran; (alpha-1,4)-D-Glucans; (alpha-1,3)-D-Glucans, e.g. pseudonigeran; Derivatives thereof
- C08B37/0021—Dextran, i.e. (alpha-1,4)-D-glucan; Derivatives thereof, e.g. Sephadex, i.e. crosslinked dextran
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Abstract
The invention discloses a method for improving the flocculation capacity of dextran. The flocculation experiment result of the invention shows that the dextran is added into the kaolin suspension firstly, and then the anti-dextran monoclonal antibody D24 is added, so that the flocculation effect of the dextran can be obviously enhanced, and the flocculation efficiency of the kaolin suspension reaches 43.2-86.5%. The invention does not need to carry out complex modification on dextran, can obviously enhance the flocculation effect of the dextran by firstly adding the dextran in a system to be flocculated and then adding the anti-dextran monoclonal antibody D24, and has the flocculation efficiency of 86.5 percent to kaolin when the final concentration of the dextran is 131.1mg/L and the final concentration of the anti-dextran monoclonal antibody is 2.4 mg/L.
Description
Technical Field
The invention belongs to the field of flocculants, and particularly relates to a method for improving the flocculation capacity of dextran.
Background
Dextran is a polysaccharide which takes glucose as a monomer and is connected with a main chain by alpha-1, 6-glycosidic bonds, and branched chains (the branched chains are glucose residues or complex glucose chains) are connected with the main chain by alpha-1, 2-, alpha-1, 3-and alpha-1, 4-glycosidic bonds. Dextran can be produced by fermenting sucrose with microorganisms, and can also be synthesized by a dextransucrase enzyme method. The molecular weight distribution is in ten-thousand to ten-million according to different generation modes and conditions. Dextran has the characteristics of large molecular weight, a large amount of hydroxyl, water solubility, flexible structure, no toxicity, biodegradability, stability in neutral environment, thermal stability, shear stability and the like, so the dextran has potential to be used as a flocculating agent. In recent years, research on biological polysaccharide flocculants is gradually started at home and abroad, but research on biological flocculants of dextran is still less.
The effect is poor when the dextran is directly used as a flocculating agent. In the existing research, dextran is generally modified, and the modified substance is used as a flocculating agent. The modified dextran has higher difficulty, and the effect is not necessarily effectively improved after modification.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provide a method for improving the flocculation capacity of dextran.
The invention aims to provide application of an anti-dextran monoclonal antibody in improving the flocculation capacity of dextran.
Preferably, the application is that after dextran is added into a system to be flocculated, the anti-dextran monoclonal antibody is added, and the mixture is stood for sedimentation.
Preferably, the final concentration of the dextran is 77.0-187.5mg/L, and the final concentration of the anti-dextran monoclonal antibody is 1.2-3.7 mg/L.
Preferably, the final concentration of the dextran is 131.1mg/L, and the final concentration of the anti-dextran monoclonal antibody is 2.4 mg/L.
Preferably, the anti-dextran monoclonal antibody is an anti-dextran monoclonal antibody D24, and the anti-dextran monoclonal antibody D24 is generated by a dextran hybridoma cell strain D24 with the preservation number of CGMCC No. 21002.
The second purpose of the invention is to provide a method for improving the flocculation capacity of dextran, which comprises the following steps: adding dextran into the flocculation system, adding anti-dextran monoclonal antibody, and standing for settling.
Preferably, the final concentration of the dextran is 77.0-187.5mg/L, and the final concentration of the anti-dextran monoclonal antibody is 1.2-3.7 mg/L.
Preferably, the final concentration of the dextran is 131.1mg/L, and the final concentration of the anti-dextran monoclonal antibody is 2.4 mg/L.
Preferably, the system to be flocculated is a kaolin suspension.
Preferably, the anti-dextran monoclonal antibody is an anti-dextran monoclonal antibody D24, and the anti-dextran monoclonal antibody D24 is generated by a dextran hybridoma cell strain D24 with the preservation number of CGMCC No. 21002.
Compared with the prior art, the invention has the following advantages and effects: the dextran can be remarkably enhanced by only adding the anti-dextran monoclonal antibody D24 in the flocculation process without carrying out complex modification on the dextran. Note that dextran and anti-dextran monoclonal antibody D24 can not be mixed in advance, and dextran must be added in the system to be flocculated first, and then anti-dextran monoclonal antibody D24 is added, and the flocculation efficiency on kaolin is as high as 86.5% when the final concentration of dextran is 131.1mg/L and the final concentration of anti-dextran monoclonal antibody is 2.4 mg/L.
The glucan hybridoma cell strain D24 is preserved in China general microbiological culture Collection center (CGMCC) in 2020, 10 months and 16 days, and has the following address: west road No. 1, north west of the morning area, beijing, 3, institute for microbiology, china academy of sciences, accession number: CGMCC No: 21002.
Detailed Description
The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.
Example 1: treating the kaolin suspension (add separately)
1. Preparation of anti-dextran monoclonal antibody D24
The method comprises the steps of inoculating 0.5mL of liquid paraffin into the abdominal cavity of a 6-8-week-old BALB/c mouse (purchased from southern medical university laboratory animal center) with strong physique and good growth condition, inoculating 5 x 10 dextran hybridoma cell strain D24 (with the preservation number of CGMCC No.21002) diluted by a serum-free culture medium (purchased from Thermo Fisher scientific) into the abdominal cavity after two weeks, and inoculating 5 x 10 dextran hybridoma cell strain D24 into the abdominal cavity of the mouse50.2 mL. After 8 days, observing the ascites generation condition of the mice every day, when the ascites is as much as possible and the mice are dying, killing the mice, and sucking the ascites out under the aseptic condition. Standing at room temperature for 30min, centrifuging at 10000rpm for 10min, collecting supernatant to obtain pretreated ascites, and freezing at-70 deg.C.
The purification scheme is an octanoic acid-ammonium sulfate precipitation method. Adding 2 parts of 0.06mol/L acetic acid buffer solution with pH4.0 into 1 part of pretreated ascites, and adjusting the pH to 4.5 by using 1mol/L HCl; adding 11 mu L of octanoic acid into diluted ascites per ml, dropwise adding octanoic acid under stirring at room temperature within 30min, standing at 4 deg.C for 2 hr, centrifuging at 12000rpm for 30min, and removing precipitate; filtering the supernatant with a nylon sieve (the diameter of the nylon sieve is 125 μm), adding 1/10 volumes of 0.1mol/L PBS buffer solution with pH7.2, and adjusting the pH to 7.2 with 1mol/L NaOH; adding saturated ammonium sulfate at 4 deg.C to 45% saturation, mixing gently for 30min, and standing for 1 hr; centrifuging at 12000rpm for 30min, and discarding the supernatant; dissolving the precipitate in 50-100 times volume of PBS buffer solution, dialyzing with 50-100 times volume of PBS, and standing overnight at 4 ℃; taking out the mixture and centrifuging the mixture for 30min at 12000rpm, removing insoluble precipitates, and freeze-drying the mixture to obtain freeze-dried powder which is named as anti-dextran monoclonal antibody D24 freeze-dried powder for later use.
2. Preparing dextran aqueous solution
Dextran T-2000(Dextran T-2000, average molecular weight 2000000, available from Shanghai leaf Biotechnology Ltd.) powder was added with water to prepare Dextran aqueous solution with concentration of 30.9, 52.6, and 75.3 mg/mL.
3. Preparing anti-dextran monoclonal antibody D24 solution
Anti-dextran monoclonal antibody D24 solutions with concentrations of 1, 2 and 3mg/mL were prepared using disodium hydrogen phosphate-sodium dihydrogen phosphate buffer (0.2mol/L, pH 7.2).
4. Flocculation experiment
A2 g/L kaolin suspension was prepared with water, 200mL of the 2g/L kaolin suspension was added to a 250mL beaker and magnetically stirred at 100 r/min. Adding 0.5mL dextran water solution, stirring for 30s, adding 0.25mL anti-dextran monoclonal antibody D24 solution, stirring for 30s, and stopping stirring. Standing for 5min, and measuring absorbance of the supernatant at 550nm (zero adjustment with distilled water). The flocculation efficiency was calculated using the following formula:
F＝(F0-F1)÷F0×100％
F0absorbance values for the blank (test 1); f1The absorbance values of the flocculation test groups.
The results are given in table 1 below:
TABLE 1 flocculation efficiency of the test groups
As can be seen from the results of the flocculation experiments (respectively) in Table 1, the dextran can be added into the kaolin suspension first, and then the anti-dextran monoclonal antibody D24 can be added, so that the flocculation effect of the dextran can be enhanced remarkably, and the flocculation efficiency of the dextran on the kaolin suspension can reach 43.2% -86.5%. Both too high and too low concentrations of the anti-dextran monoclonal antibody D24 significantly affected the flocculation efficiency of dextran at the same dextran concentration.
Example 2: treating kaolin suspension (adding after mixing)
1. Preparation of anti-dextran monoclonal antibody D24
The method comprises the steps of inoculating 0.5mL of liquid paraffin into the abdominal cavity of a 6-8-week-old BALB/c mouse (purchased from southern medical university laboratory animal center) with strong physique and good growth condition, inoculating 5 x 10 dextran hybridoma cell strain D24 (with the preservation number of CGMCC No.21002) diluted by a serum-free culture medium (purchased from Thermo Fisher scientific) into the abdominal cavity after two weeks, and inoculating 5 x 10 dextran hybridoma cell strain D24 into the abdominal cavity of the mouse50.2 mL. After 8 days, observing the ascites generation condition of the mice every day, when the ascites is as much as possible and the mice are dying, killing the mice, and sucking the ascites out under the aseptic condition. Standing at room temperature for 30min, centrifuging at 10000rpm for 10min, collecting supernatant to obtain pretreated ascites, and freezing at-70 deg.C.
The purification scheme is an octanoic acid-ammonium sulfate precipitation method. Adding 2 parts of 0.06mol/L acetic acid buffer solution with pH4.0 into 1 part of pretreated ascites, and adjusting the pH to 4.5 by using 1mol/L HCl; adding 11 mu L of octanoic acid into diluted ascites per ml, dropwise adding octanoic acid under stirring at room temperature within 30min, standing at 4 deg.C for 2 hr, centrifuging at 12000rpm for 30min, and removing precipitate; filtering the supernatant with a nylon sieve (the diameter of the nylon sieve is 125 μm), adding 1/10 volumes of 0.1mol/L PBS buffer solution with pH7.2, and adjusting the pH to 7.2 with 1mol/L NaOH; adding saturated ammonium sulfate at 4 deg.C to 45% saturation, mixing gently for 30min, and standing for 1 hr; centrifuging at 12000rpm for 30min, and discarding the supernatant; dissolving the precipitate in 50-100 times volume of PBS buffer solution, dialyzing with 50-100 times volume of PBS, and standing overnight at 4 ℃; taking out the mixture and centrifuging the mixture for 30min at 12000rpm, removing insoluble precipitates, and freeze-drying the mixture to obtain freeze-dried powder which is named as anti-dextran monoclonal antibody D24 freeze-dried powder for later use.
2. Preparing dextran aqueous solution
Dextran T-2000(Dextran T-2000, average molecular weight 2000000, available from Shanghai leaf Biotechnology Ltd.) powder was added with water to prepare Dextran aqueous solution with concentration of 30.9, 52.6, and 75.3 mg/mL.
3. Preparing anti-dextran monoclonal antibody D24 solution
Anti-dextran monoclonal antibody D24 solutions with concentrations of 1, 2 and 3mg/mL were prepared using disodium hydrogen phosphate-sodium dihydrogen phosphate buffer (0.2mol/L, pH 7.2).
4. Preparation of flocculant
According to the dextran water solution: the volume ratio of the anti-dextran monoclonal antibody D24 solution is 2: 1, preparing a flocculating agent.
5. Flocculation experiment
A2 g/L kaolin suspension was prepared with water, 200mL of the 2g/L kaolin suspension was added to a 250mL beaker and magnetically stirred at 100 r/min. 0.75mL of flocculant was added, and after stirring for 1min, the stirring was stopped. Standing for 5min, and measuring absorbance of the supernatant at 550nm (zero adjustment with distilled water). The flocculation efficiency was calculated using the following formula:
F＝(F0-F1)÷F0×100％
F0absorbance values for the blank (test 1); f1The absorbance values of the flocculation test groups.
The results are given in table 2 below:
TABLE 2 flocculation efficiency of the test groups
As can be seen from the results of the flocculation experiments (adding after mixing) in Table 2, the flocculation efficiency of adding the kaolin suspension after the dextran and the anti-dextran monoclonal antibody D24 are pre-mixed to prepare the flocculant is only 4.7% -15.3%, so that the flocculant is ineffective when the dextran and the anti-dextran monoclonal antibody D24 are pre-mixed and then added into the flocculation system.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. The application of the anti-dextran monoclonal antibody in improving the flocculation capacity of dextran.
2. The use of claim 1, wherein the use comprises adding dextran into the flocculation system, adding anti-dextran monoclonal antibody, and standing for sedimentation.
3. The use of claim 2, wherein the final dextran concentration is 77.0-187.5mg/L, and the final anti-dextran monoclonal antibody concentration is 1.2-3.7 mg/L.
4. The use of claim 3, wherein the final dextran concentration is 131.1mg/L and the final anti-dextran monoclonal antibody concentration is 2.4 mg/L.
5. The use of any one of claims 1-4, wherein said anti-dextran monoclonal antibody is anti-dextran monoclonal antibody D24, and said anti-dextran monoclonal antibody D24 is produced by dextran hybridoma cell line D24 with accession number CGMCC No. 21002.
6. A method for improving flocculation capacity of dextran, which is characterized by comprising the following steps: adding dextran into the flocculation system, adding anti-dextran monoclonal antibody, and standing for settling.
7. The method of claim 6, wherein the final concentration of dextran is 77.0-187.5mg/L, and the final concentration of the anti-dextran monoclonal antibody is 1.2-3.7 mg/L.
8. The method of claim 7, wherein the final dextran concentration is 131.1mg/L and the final anti-dextran monoclonal antibody concentration is 2.4 mg/L.
9. The method of claim 6, wherein the system to be flocculated is a kaolin suspension.
10. The method for improving the flocculation capacity of dextran according to any of claims 6-9, wherein said anti-dextran monoclonal antibody is anti-dextran monoclonal antibody D24, and said anti-dextran monoclonal antibody D24 is produced by dextran hybridoma cell strain D24 with the preservation number of CGMCC No. 21002.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116649458A (en) * | 2023-06-26 | 2023-08-29 | 广东省科学院生物与医学工程研究所 | Method for improving gel performance of soybean protein isolate copolymer |
| CN116813458A (en) * | 2023-06-26 | 2023-09-29 | 广东省科学院生物与医学工程研究所 | Method for improving loading performance of bovine serum albumin copolymer |
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| EP3141558A1 (en) * | 2015-09-08 | 2017-03-15 | Merck Patent GmbH | Affinity chromatography media for removal of anti-a and/or anti-b antibodies |
| CN110437378A (en) * | 2019-09-09 | 2019-11-12 | 安徽春霖环保科技有限公司 | Cationic dextran based polyalcohol, preparation and the application as flocculant |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116649458A (en) * | 2023-06-26 | 2023-08-29 | 广东省科学院生物与医学工程研究所 | Method for improving gel performance of soybean protein isolate copolymer |
| CN116813458A (en) * | 2023-06-26 | 2023-09-29 | 广东省科学院生物与医学工程研究所 | Method for improving loading performance of bovine serum albumin copolymer |
| CN116649458B (en) * | 2023-06-26 | 2024-03-12 | 广东省科学院生物与医学工程研究所 | Method for improving gel performance of soybean protein isolate copolymer |
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Address after: No.10, shiliugang Road, Haizhu District, Guangzhou City, Guangdong Province 510000 Patentee after: Institute of biological and medical engineering, Guangdong Academy of Sciences Address before: No.10, shiliugang Road, Haizhu District, Guangzhou City, Guangdong Province 510000 Patentee before: Institute of bioengineering, Guangdong Academy of Sciences |