CN103897196A - Polymer and preparation method thereof - Google Patents
Polymer and preparation method thereof Download PDFInfo
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- CN103897196A CN103897196A CN201210578582.2A CN201210578582A CN103897196A CN 103897196 A CN103897196 A CN 103897196A CN 201210578582 A CN201210578582 A CN 201210578582A CN 103897196 A CN103897196 A CN 103897196A
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Abstract
The invention is applicable to the field of high polymer materials and provides a polymer and a preparation method thereof. The preparation method for the polymer comprises the following steps: preparation of a first intermediate; preparation of a second intermediate; preparation of a polymer with a carboxyl group; preparation of the polymer; etc. The polymer has poly(2-methyl-2-oxazoline) and other groups, has high biological inertness, can both specifically bind to certain proteins and block non-specific bonding of other macro-molecular substances on the surface of the polymer and is applicable to accurate, sensitive, quantitative and highly efficient immunological detection. According to the preparation method for the polymer, the polymer having poly(2-methyl-2-oxazoline) and other groups is prepared through polymerization, so the polymer has high biological inertness, can both specifically bind to certain proteins and block non-specific bonding of other macro-molecular substances on the surface of the polymer and is applicable to accurate, sensitive, quantitative and highly efficient immunological detection.
Description
Technical field
The invention belongs to field of new, relate in particular to a kind of polymkeric substance and preparation method thereof.
Background technology
In recent years, blood circulation tumour cell be detected as for biomedical sector pay close attention to a focus.As everyone knows, the generation evolution of cancer generally will be experienced following process.First be formerly to send out generation carcinoid, next is the vasculogenesis of tumor tissues, then be the formation of primary malignant neoplasm, be that malignant cell is invaded blood vessel by basilar membrane and becomes blood circulation tumour cell again, last malignant cell is invaded remaining healthy tissues by vessel wall and forms second malignant neoplasm.Once second malignant neoplasm forms, excision has just lost meaning, and it is very difficult that the treatment of cancer becomes.Under normal circumstances, the circulating tumor cell in blood can stop the longer time in blood, just can invade remaining healthy tissues and form second malignant neoplasm.Therefore, the detection of blood circulation tumour cell becomes an important indicator of cancer diagnosis and definite treatment plan gradually.In addition, by the monitoring to blood circulation tumour cell concentration in cancer patient, can also assess the curative effect of cancer therapy drug.Due to the concentration of circulating tumor cell in blood very little (every 100,000 to 1,000,000,000 blood cell kinds have one), therefore the screening of circulating tumor cell becomes a challenging problem.Current major programme is to utilize the volume size difference of circulating tumor cell and normal blood cell and circulating tumor cell and some antibody to be used for sieving as the specific binding of anti-epithelial cell adhesion molecule, antikeratin antibody etc.For first scheme, needing the problem solving is that preparation can some biomacromolecule of specific binding, can stop the material of all the other biomacromolecule non-specific binding simultaneously.In recent years, immunology detection chip becomes the focus that biomedical sector is paid close attention to.Accurately, sensitive, quantitative, efficient immunology detection is the target that people pursue always.Equally, in order to reach above requirement, immunology chip need to possess can some protein molecular of specific binding, the surface that can stop all the other biomacromolecule non-specific binding simultaneously.
At present, the material of this respect is actually rare.
Summary of the invention
In view of this, the object of the present invention is to provide the one can some biomacromolecule of specific binding, can stop the macromolecular material of all the other biomacromolecule non-specific binding simultaneously.
The present invention is achieved in that
A kind of polymkeric substance, has following structural formula:
This x, y, z is all selected from the natural number of 1-500, and the x+y+z natural number that is 50-500.
And above-mentioned method for producing polymer, comprises the steps:
2-methyl-2-oxazoline and initiator are added in organic solvent, be to react 6-48 hour under 60-80 DEG C of condition in temperature, temperature is adjusted to room temperature, add terminator, stirring reaction 2-24 hour, solvent in solution after reaction is removed, stirring adds water, be 9-14 Water Under solution 2-36 hour by the reaction product adding water after stirring in pH value, pH after hydrolysis is adjusted to 6-8, dry first intermediate product that obtains of dialysis, this initiator is selected from 3-ethyl bromide or 3-methyl bromide c, this terminator be selected from two ends all with amino polymkeric substance as quadrol, butanediamine, hexanediamine,
By first solution that obtains soluble in water this first intermediate product, folic acid or vitamin H are dissolved in dimethyl sulfoxide (DMSO) or dimethyl formamide and add N-(3-dimethylamino-propyl)-N '-ethyl-carbodiimide hydrochloride stirs and obtains the second solution, this second solution is added in the first solution, at room temperature react 2-48 hour, solution dialysis after reaction is obtained to the second intermediate product;
2-methyl-2-oxazoline and initiator are added in organic solvent, be to react 6-48 hour under 60-80 DEG C of condition in temperature, temperature is adjusted to room temperature, add terminator, stirring reaction 2-24 hour, solvent after reaction is removed, stirring adds water, be 9-14 Water Under solution 2-36 hour by the reaction product adding water after stirring in pH value, pH after hydrolysis is adjusted to 6-8, the dry polymkeric substance obtaining with ester group of dialysis, this initiator is selected from trifluoromethanesulfonic acid methyl esters or methyl iodide, this terminator is selected from an end with primary amine or secondary amine, another end is selected from 4-piperidine carboxylate with the polymkeric substance of ester group, 4-piperidine carboxylic acid methyl esters, glycine ethyl ester, glycine methyl ester, 3-alanine ethyl ester, 3-alanine methyl esters etc.,
The bromate of PLL is added in damping fluid, add this polymkeric substance and this second intermediate product with carboxyl, add N-(3-dimethylamino-propyl)-N '-ethyl-carbodiimide hydrochloride and N-hydroxy-succinamide sulfonate sodium, react at ambient temperature 1-32 hour, after reacting, solution dialysis is dried, and obtains polymkeric substance.
Polymkeric substance of the present invention, there are poly-(2-methyl-2-oxazoline) and other groups, there is very high biologically inert, can either some protein of specific binding, can stop that remaining macromolecular substance non-specific binding, on its surface, can be used in accurate, sensitive, quantitative and efficient immunology detection simultaneously; Polymerization process for preparing of the present invention, obtain having the polymkeric substance of poly-(2-methyl-2-oxazoline) and other groups by polyreaction, realize this polymkeric substance and there is very high biologically inert, can either some protein of specific binding, can stop that remaining macromolecular substance non-specific binding, on its surface, can be used in accurate, sensitive, quantitative and efficient immunology detection simultaneously.
Brief description of the drawings
Fig. 1 is polymkeric substance prepared by the embodiment of the present invention one
1h-nuclear magnetic spectrum.
Embodiment
In order to make object of the present invention, technical scheme and advantage clearer, below in conjunction with drawings and Examples, the present invention is further elaborated.Should be appreciated that specific embodiment described herein, only in order to explain the present invention, is not intended to limit the present invention.
The embodiment of the present invention provides a kind of polymkeric substance, and this polymkeric substance has following structural formula:
This x, y, z is all selected from the natural number of 1-500, and the x+y+z natural number that is 50-500.This polymkeric substance can be regarded the multipolymer of three kinds of repeating units of x, y, z representative as, is random copolymers, but not segmented copolymer.
Polymkeric substance of the present invention, there are poly-(2-methyl-2-oxazoline) and other groups, there is very high biologically inert, can either some protein of specific binding, can stop that remaining macromolecular substance non-specific binding, on its surface, can be used in accurate, sensitive, quantitative and efficient immunology detection simultaneously.
The embodiment of the present invention further provides above-mentioned method for producing polymer, comprises the steps:
Step S01, preparation the first intermediate product:
2-methyl-2-oxazoline and initiator are added in organic solvent, be to react 6-48 hour under 60-80 DEG C of condition in temperature, temperature is adjusted to room temperature, add terminator, stirring reaction 2-24 hour, solvent in solution after reaction is removed, stirring adds water, be 9-14 Water Under solution 2-36 hour by the reaction product adding water after stirring in pH value, pH after hydrolysis is adjusted to 6-8, dry first intermediate product that obtains of dialysis, this initiator is selected from 3-ethyl bromide or 3-methyl bromide c, this terminator is selected from two ends all with amino polymkeric substance, these two ends all with amino polymkeric substance as quadrol, butanediamine or hexanediamine etc.,
Step S02, preparation the second intermediate product:
By first solution that obtains soluble in water this first intermediate product, folic acid or vitamin H are dissolved in dimethyl sulfoxide (DMSO) or dimethyl formamide and add N-(3-dimethylamino-propyl)-N '-ethyl-carbodiimide hydrochloride stirs and obtains the second solution, this second solution is added in the first solution, at room temperature react 2-48 hour, solution dialysis after reaction is obtained to the second intermediate product;
Step S03, the polymkeric substance of preparation with carboxyl:
2-methyl-2-oxazoline and initiator are added in organic solvent, be to react 6-48 hour under 60-80 DEG C of condition in temperature, temperature is adjusted to room temperature, add terminator, stirring reaction 2-24 hour, solvent after reaction is removed, stirring adds water, be 9-14 Water Under solution 2-36 hour by the reaction product adding water after stirring in pH value, pH after hydrolysis is adjusted to 6-8, the dry polymkeric substance obtaining with carboxyl of dialysis, this initiator is selected from trifluoromethanesulfonic acid methyl esters or methyl iodide, this terminator is selected from an end with primary amine or secondary amine, another end is with the polymkeric substance of ester group, for example be selected from 4-piperidine carboxylate, 4-piperidine carboxylic acid methyl esters, glycine ethyl ester, glycine methyl ester, 3-alanine ethyl ester, 3-alanine methyl esters etc.
S04, prepares polymkeric substance:
The bromate of PLL is added in damping fluid, add this polymkeric substance and this second intermediate product with carboxyl, add N-(3-dimethylamino-propyl)-N '-ethyl-carbodiimide hydrochloride and N-hydroxy-succinamide sulfonate sodium, react at ambient temperature 1-32 hour, after reacting, solution dialysis is dried, and obtains polymkeric substance.
In step S01, the mol ratio of this initiator and 2-methyl-2-oxazoline is 0.8-4:100, and the mol ratio of this terminator and 2-methyl-2-oxazoline is 4-100:100.This organic solvent is selected from acetonitrile or dimethyl formamide.
Step S01 is specific as follows:
After 2-methyl-2-oxazoline and initiator for reaction are completed, system temperature is adjusted to room temperature, for example 20-30 DEG C adds terminator in reaction system, then stirs 2-24 hour.Solvent after reaction is evaporated under condition of negative pressure, the organic solvent reacting in rear solution is removed, obtained oily thick liquid, in this oily thick liquid, add water, stir, obtaining mixing solutions, is 9-14 Water Under solution 2-36 hour by this mixing solutions in pH value, and pH after hydrolysis is adjusted to 6-8, the solution of adjusting pH to 6-8 is inserted in dialysis tubing, dialysis treatment in deionized water, the time more than 3 days, obtains the first intermediate product.
In step S02, the mol ratio of this first intermediate product and described folic acid or vitamin H is 1:2-5.Step S02 is specific as follows:
By soluble in water this first intermediate product, stir and obtain the first solution;
Then folic acid or vitamin H are dissolved in dimethyl sulfoxide (DMSO) or dimethyl formamide and add N-(3-dimethylamino-propyl)-N '-ethyl-carbodiimide hydrochloride stirs and obtains the second solution;
This second solution is added in the first solution, at room temperature stirs, reaction 2-48 hour, proceeds to solution after reaction in dialysis tubing, dialyses more than 3 days in deionized water, by the solution lyophilize that contains polymkeric substance, obtains the second intermediate product;
In step S03, the mol ratio of this initiator and described 2-methyl-2-oxazoline is 0.8-4:100; The mol ratio of this terminator and 2-methyl-2-oxazoline is 4-100:100.Identical in this organic solvent and step S01, do not repeat to set forth at this.
Step S03 is specific as follows:
After 2-methyl-2-oxazoline and initiator for reaction are completed, system temperature is adjusted to room temperature, for example 20-30 DEG C adds terminator in reaction system, then stirs 2-24 hour.Solvent after reaction is evaporated under condition of negative pressure, the organic solvent reacting in rear solution is removed, obtain oily thick liquid, in this oily thick liquid, add water, stir, obtain mixing solutions, be 9-14 Water Under solution 2-36 hour by this mixing solutions in pH value, pH after hydrolysis is adjusted to 6-8, the solution that pH is adjusted to 6-8 is inserted in dialysis tubing, dialysis treatment in deionized water, obtains carboxylic polymkeric substance;
In step S04, the mol ratio of the bromate of this PLL and the described polymkeric substance with carboxyl is 1:0.002-0.5; The mol ratio of the bromate of this PLL and described the second intermediate product is 1:0.002-0.5; The bromate of this PLL and N-(3-dimethylamino-propyl) mol ratio of-N '-ethyl-carbodiimide hydrochloride is 1:0.02-10, the bromate of this PLL and with the mol ratio of N-hydroxy-succinamide sulfonate sodium be 1:0.004-0.5.This damping fluid is preferably 0.01mol/L, the phosphate buffered saline buffer of pH=7.4.
In step S04, after having reacted, solution after reaction is transferred in dialysis tubing, in phosphate buffered saline buffer and deionized water, dialyse respectively more than 3 days, the liquid freezing that contains polymkeric substance is dry, obtain polymkeric substance, this phosphate buffered saline buffer and aforementioned identical, does not repeat to set forth at this.
Polymerization process for preparing of the present invention, obtain having the polymkeric substance of poly-(2-methyl-2-oxazoline) and other groups by polyreaction, realize this polymkeric substance and there is very high biologically inert, can either some protein of specific binding, can stop that remaining macromolecular substance non-specific binding, on its surface, can be used in accurate, sensitive, quantitative and efficient immunology detection simultaneously.
Below in conjunction with specific embodiment, above-mentioned method for producing polymer is described in detail.
Embodiment mono-
The chemical structural formula of embodiment of the present invention polymkeric substance is as follows:
Embodiment of the present invention method for producing polymer, comprises the steps:
1, two ends are respectively with carboxyl and amino gathering synthesizing of (2-methyl-2-oxazoline):
2-methyl-2-oxazoline (8.5mL, 100mmol) is dissolved in to acetonitrile (30mL), then 3-ethyl bromide (0.520mL, 4mmol) is joined in reaction system, mixture is heated to 70 DEG C afterwards, react 8 hours.Stop subsequently heating, thing to be mixed is cooled to room temperature, adds quadrol (6.65mL, 100mmol) to stir 12 hours.Under negative pressure, with heating, partial solvent is removed, obtained oily thick liquid.Add deionized water to 50mL, the Water Under solution of pH=14 24 hours, pH value of solution is adjusted to neutrality, then the solution that contains polymkeric substance is transferred in dialysis tubing, in deionized water, dialyse 24 hours.Finally, by the solution lyophilize that contains polymkeric substance or decompression heat drying, obtain poly-(2-methyl-2-oxazoline) white solid that molecular weight is about 2K, structural formula is as follows:
2, modified with folic acid poly-(2-methyl-2-oxazoline) is synthetic:
Two ends obtained above are dissolved in deionized water (6.0mL) to prepare the aqueous solution of polymkeric substance with carboxyl and amino poly-(2-methyl-2-oxazoline) white solid (2K, 0.20g, 0.1mmol) respectively.Again folic acid (132mg, 0.3mmol) is dissolved in dimethyl sulfoxide (DMSO) (4.0mL), adds afterwards N-(3-dimethylamino-propyl)-N '-ethyl-carbodiimide hydrochloride (577mg, 3.0mmol), be stirred to completely and dissolve.Then the solution containing folic acid is all added in polymers soln, at room temperature react 48 hours.After reaction finishes, the solution that contains polymkeric substance is transferred in dialysis tubing, in deionized water, dialysed 7 days.Finally, by the solution lyophilize that contains polymkeric substance, obtain poly-(2-methyl-2-oxazoline) yellow solid of modified with folic acid that molecular weight is about 2.5K, structural formula is as follows:
3,2-methyl-2-oxazoline (8.5mL, 100mmol) is dissolved in to acetonitrile (30mL), then trifluoromethanesulfonic acid methyl esters (0.44mL, 4mmol) is joined in reaction system, mixture is heated to 70 DEG C afterwards, react 8 hours.Stop subsequently heating, thing to be mixed is cooled to room temperature, adds 4-piperidine carboxylate (1.224mL, 8mmol) to stir 12 hours.Under negative pressure, with heating, partial solvent is removed, obtained oily thick liquid.Add deionized water to 50mL, the Water Under solution of pH=14 24 hours, pH value of solution is adjusted to neutrality, then the solution that contains polymkeric substance is transferred in dialysis tubing, in deionized water, dialyse 24 hours.Finally, by the solution lyophilize that contains polymkeric substance or decompression heat drying, obtaining molecular weight is poly-(2-methyl-2-oxazoline) white solid of 2K, and structural formula is as follows:
4, synthesizing of PLL-grafting-poly-(2-methyl-2-oxazoline) modified with folic acid poly-(2-methyl-2-oxazoline):
By the bromate of PLL (molecular weight 21K, 21mg, 1 μ mol) be dissolved in 3mL phosphate-buffered salt (0.01mol/L, pH=7.4) in solution, adding molecular weight is 2K, simple function reunion (2-methyl-2-oxazoline) (48mg, 24 μ mol) of end band carboxyl prepared by step 3 and poly-(2-methyl-2-oxazoline) (2.5K of modified with folic acid obtained above, 15mg, 6 μ mol).Add again N-(3-dimethylamino-propyl)-N '-ethyl-carbodiimide hydrochloride (57.6mg, 300 μ mol) and N-hydroxy-succinamide sulfonate sodium (6.51mg, 30 μ mol) react 16 hours under room temperature.After reaction finishes, the solution that contains polymkeric substance is transferred in dialysis tubing, in phosphate-buffered salt (0.01mol/L, pH=7.4) solution and deionization, dialysed 7 days respectively.Finally, by the solution lyophilize that contains polymkeric substance, obtaining molecular weight is 80K PLL-grafting-poly-(2-methyl-2-oxazoline) poly-(2-methyl-2-oxazoline) yellow solid of modified with folic acid (grafting density 0.3, wherein modified with folic acid poly-(2-methyl-2-oxazoline) accounts for 20%).
Refer to Fig. 1, Fig. 1 shows the polymkeric substance nuclear magnetic spectrogram that the embodiment of the present invention is prepared, and as can be seen from Figure 1, the prepared polymkeric substance of above-mentioned preparation method is the polymkeric substance that said structure formula characterizes.
The polymkeric substance obtaining that the embodiment of the present invention one is prepared, can be used for preparing the biochip that detects circulating tumor cell in blood.Transition metal oxide as tantalum pentoxide, Niobium Pentxoxide, titanium dioxide, indium tin oxide target and silicon-dioxide etc. at physiological condition lower surface with negative charge, PLL is with positive charge.The dilute solution of poly-PLL-grafting-modified with folic acid (2-methyl-2-oxazoline) is dropped in to the silica chip of processing through Piranha solution oxide or be coated with the silica chip surface of above various transition metal oxides, PLL-grafting-modified with folic acid poly-(2-methyl-2-oxazoline) can spontaneously be adsorbed onto electronegative surface under electrostatic interaction, forms unimolecular layer.Because poly-(2-methyl-2-oxazoline) can stop the non-specific adsorption of biomolecules, simultaneously fixing folate molecule can with the folacin receptor specific binding of overexpression on tumor cell membrane, the chip of being prepared by this material can optionally adsorb the circulating tumor cell in blood.In addition, the technology such as binding immunoassay fluorescent staining method, polymerase chain reaction, optical waveguide optical spectroscopy, can carry out molecular biology or physicochemical property are analyzed to the circulating tumor cancer cells sieving out.
Embodiment bis-
The chemical structural formula of embodiment of the present invention polymkeric substance is as follows:
Embodiment of the present invention method for producing polymer, comprises the steps:
1, two ends are respectively with carboxyl and amino gathering synthesizing of (2-methyl-2-oxazoline):
2-methyl-2-oxazoline (8.5mL, 100mmol) is dissolved in to acetonitrile (30mL), then 3-ethyl bromide (0.520mL, 4mmol) is joined in reaction system, mixture is heated to 70 DEG C afterwards, react 8 hours.Stop subsequently heating, thing to be mixed is cooled to room temperature, adds quadrol (6.65mL, 100mmol) to stir 12 hours.Under negative pressure, with heating, partial solvent is removed, obtained oily thick liquid.Add deionized water to 50mL, the Water Under solution of pH=14 24 hours, pH value of solution is adjusted to neutrality, then the solution that contains polymkeric substance is transferred in dialysis tubing, in deionized water, dialyse 24 hours.Finally, by the solution lyophilize that contains polymkeric substance or decompression heat drying, obtain poly-(2-methyl-2-oxazoline) white solid that molecular weight is about 2K, structural formula is as follows:
2, biotin modification poly-(2-methyl-2-oxazoline) is synthetic:
Two ends obtained above are dissolved in deionized water (6.0mL) to prepare the aqueous solution of polymkeric substance with carboxyl and amino poly-(2-methyl-2-oxazoline) white solid (2K, 0.20g, 0.1mmol) respectively.Again vitamin H (73mg, 0.3mmol) is dissolved in dimethyl sulfoxide (DMSO) (4.0mL), adds afterwards N-(3-dimethylamino-propyl)-N '-ethyl-carbodiimide hydrochloride (577mg, 3.0mmol), be stirred to completely and dissolve.Then the solution containing vitamin H is all added in polymers soln, at room temperature react 48 hours.After reaction finishes, the solution that contains polymkeric substance is transferred in dialysis tubing, in deionized water, dialysed 7 days.Finally, by the solution lyophilize that contains polymkeric substance, obtain poly-(2-methyl-2-oxazoline) white solid of biotin modification that molecular weight is about 2.2K, structural formula is as follows:
3,2-methyl-2-oxazoline (8.5mL, 100mmol) is dissolved in to acetonitrile (30mL), then trifluoromethanesulfonic acid methyl esters (0.44mL, 4mmol) is joined in reaction system, mixture is heated to 70 DEG C afterwards, react 8 hours.Stop subsequently heating, thing to be mixed is cooled to room temperature, adds 4-piperidine carboxylate (1.224mL, 8mmol) to stir 12 hours.Under negative pressure, with heating, partial solvent is removed, obtained oily thick liquid.Add deionized water to 50mL, the Water Under solution of pH=14 24 hours, pH value of solution is adjusted to neutrality, then the solution that contains polymkeric substance is transferred in dialysis tubing, in deionized water, dialyse 24 hours.Finally, by the solution lyophilize that contains polymkeric substance or decompression heat drying, obtaining molecular weight is poly-(2-methyl-2-oxazoline) white solid of 2K, and structural formula is as follows:
4, synthesizing of PLL-grafting-poly-(2-methyl-2-oxazoline) biotin modification poly-(2-methyl-2-oxazoline):
By the bromate of PLL (molecular weight 21K, 21mg, 1 μ mol) be dissolved in 3mL phosphate-buffered salt (0.01mol/L, in pH=7.4 solution, adding molecular weight is 2K, simple function reunion (2-methyl-2-oxazoline) (48mg, 24 μ mol) of end band carboxyl prepared by step 3 and poly-(2-methyl-2-oxazoline) (2.2K of biotin modification obtained above, 13.2mg, 6 μ mol).Add again N-(3-dimethylamino-propyl)-N '-ethyl-carbodiimide hydrochloride (57.6mg, 300 μ mol) and N-hydroxy-succinamide sulfonate sodium (6.51mg, 30 μ mol) react 16 hours under room temperature.After reaction finishes, the solution that contains polymkeric substance is transferred in dialysis tubing, in phosphate-buffered salt (0.01mol/L, pH=7.4) solution and deionized water, dialysed 7 days respectively.Finally, by the solution lyophilize that contains polymkeric substance, obtaining molecular weight is 80K PLL-grafting-poly-(2-methyl-2-oxazoline) poly-(2-methyl-2-oxazoline) white solid of biotin modification (grafting density 0.3, wherein biotin modification poly-(2-methyl-2-oxazoline) accounts for 20%).
The nuclear magnetic spectrogram of the prepared polymkeric substance of the embodiment of the present invention two and aforesaid similar, does not repeat to set forth at this.This material of this polymkeric substance can be used for preparation and detects the biochip of antigen in blood or antibody protein.Transition metal oxide as tantalum pentoxide, Niobium Pentxoxide, titanium dioxide, indium tin oxide target and silicon-dioxide etc. at physiological condition lower surface with negative charge, PLL is with positive charge.The dilute solution of poly-PLL-grafting-biotin modification (2-methyl-2-oxazoline) is dropped in to the silica chip of processing through Piranha solution oxide or be coated with the silica chip surface of above various transition metal oxides, PLL-grafting-biotin modification poly-(2-methyl-2-oxazoline) can spontaneously be adsorbed onto electronegative surface under electrostatic interaction, forms unimolecular layer.Due to fixing biotin molecule can with streptavidin selective binding, poly-(2-methyl-2-oxazoline) can stop the non-specific adsorption of biomolecules simultaneously, the antibody that streptavidin is modified or antigen protein can be by being fixed to the affinity interaction of fixing vitamin H the chip surface of being prepared by this material, thereby can optionally adsorb antigen or antibody protein corresponding in blood.The technology such as binding immunoassay fluorescent staining method, optical waveguide optical spectroscopy, can carry out accurate, sensitive, quantitative, analysis efficiently to various antigen or antibody protein in blood.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any amendments of doing within the spirit and principles in the present invention, be equal to and replace and improvement etc., within all should being included in protection scope of the present invention.
Claims (9)
1. a polymkeric substance, has following structural formula:
Described x, y, z is all selected from the natural number of 1-500, and the x+y+z natural number that is 50-500.
2. method for producing polymer as claimed in claim 1, comprises the steps:
2-methyl-2-oxazoline and initiator are added in organic solvent, be to react 6-48 hour under 60-80 DEG C of condition in temperature, temperature is adjusted to room temperature, add terminator, stirring reaction 2-24 hour, solvent in solution after reaction is removed, stirring adds water, be 9-14 Water Under solution 2-36 hour by the reaction product adding water after stirring in pH value, pH after hydrolysis is adjusted to 6-8, dry first intermediate product that obtains of dialysis, described initiator is selected from 3-ethyl bromide or 3-methyl bromide c, described terminator is selected from two ends all with amino polymkeric substance,
By first solution that obtains soluble in water described the first intermediate product, folic acid or vitamin H are dissolved in dimethyl sulfoxide (DMSO) or dimethyl formamide and add N-(3-dimethylamino-propyl)-N '-ethyl-carbodiimide hydrochloride, stirring obtains the second solution, described the second solution is added in the first solution, at room temperature react 2-48 hour, by dry solution dialysis after reaction second intermediate product that obtains;
2-methyl-2-oxazoline and initiator are added in organic solvent, be to react 6-48 hour under 60-80 DEG C of condition in temperature, temperature is adjusted to room temperature, add terminator, stirring reaction 2-24 hour, solvent after reaction is removed, stirring adds water, be 9-14 Water Under solution 2-36 hour by the reaction product adding water after stirring in pH value, pH after hydrolysis is adjusted to 6-8, the dry polymkeric substance obtaining with carboxyl of dialysis, described initiator is selected from trifluoromethanesulfonic acid methyl esters or methyl iodide, described terminator is selected from an end with primary amine or secondary amine, another end is with the polymkeric substance of ester group,
The bromate of PLL is added in damping fluid, add described polymkeric substance and described the second intermediate product with carboxyl, add N-(3-dimethylamino-propyl)-N '-ethyl-carbodiimide hydrochloride and N-hydroxy-succinamide sulfonate sodium, react at ambient temperature 1-32 hour, after reacting, solution dialysis is dried, and obtains polymkeric substance.
3. method for producing polymer as claimed in claim 2, is characterized in that, prepare in described the first intermediate product step, the mol ratio of described initiator and described 2-methyl-2-oxazoline is 0.8-4:100.
4. method for producing polymer as claimed in claim 2, is characterized in that, prepare in described the first intermediate product step, the mol ratio of described terminator and described 2-methyl-2-oxazoline is 4-100:100.
5. method for producing polymer as claimed in claim 2, is characterized in that, the mol ratio of described the first intermediate product and described folic acid or vitamin H is 1:2-5.
6. method for producing polymer as claimed in claim 2, is characterized in that, the mol ratio of the bromate of described PLL and the described polymkeric substance with carboxyl is 1:0.002-0.5.
7. method for producing polymer as claimed in claim 2, is characterized in that, the mol ratio of the bromate of described PLL and described the second intermediate product is 1:0.002-0.5.
8. method for producing polymer as claimed in claim 2, it is characterized in that, prepare in the described polymkeric substance step with carboxyl, a described end is with primary amine or secondary amine, another end is selected from 4-piperidine carboxylate with the polymkeric substance of ester group, 4-piperidine carboxylic acid methyl esters, glycine ethyl ester, glycine methyl ester, 3-alanine ethyl ester or 3-alanine methyl esters.
9. method for producing polymer as claimed in claim 2, is characterized in that, prepare in the described polymkeric substance step with carboxyl, the mol ratio of described initiator and described 2-methyl-2-oxazoline is 0.8-4:100; The mol ratio of described terminator and described 2-methyl-2-oxazoline is 4-100:100.
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|---|---|---|---|---|
| CN102604083A (en) * | 2011-01-18 | 2012-07-25 | 中国医学科学院医药生物技术研究所 | Polymer modified lipid material and application thereof |
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2012
- 2012-12-27 CN CN201210578582.2A patent/CN103897196B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102604083A (en) * | 2011-01-18 | 2012-07-25 | 中国医学科学院医药生物技术研究所 | Polymer modified lipid material and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| RUPERT KONRADI ET AL.: "Poly-2-methyl-2-oxazoline: a peptide-like polymer for protein-repellent sulfaces", 《LANGMUIR》 * |
| THOMAS VON ERLACH ET AL.: "Formation and characterization of DNA-polymer-condensates based on poly(2-methyl-2-oxazoline) grafted poly(L-lysine) for non-viral delivery of therapeutic DNA", 《BIOMATERIALS》 * |
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| CN103897196B (en) | 2016-12-28 |
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