Method for testing interaction of temperature-sensitive random polymer with agglutinin

A determination method and polymer technology, which can be used in the preparation of test samples, measurement of scattering properties, measurement of color/spectral properties, etc., can solve the problems of unclear identification process between sugar-based polymers and lectins, and achieve fast test analysis. , Simple operation, low cost effect

Inactive Publication Date: 2019-10-29
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a method for measuring the interaction between thermosensitive random polymers and lectins, so as to overcome the defect that the recognition process between sugar-based polymers and lectins is not clear in the prior art

Method used

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  • Method for testing interaction of temperature-sensitive random polymer with agglutinin
  • Method for testing interaction of temperature-sensitive random polymer with agglutinin
  • Method for testing interaction of temperature-sensitive random polymer with agglutinin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Preparation of P(DEGMA-co-OVNGlu): DDATC was used as chain transfer agent and ACCN was used as initiator. Take 0.3313g DEGMA, 0.1470g OVNGlu, 8.08mg DDATC, 0.305mg ACCN dissolved in 1.5mL DMF solution, after 3 times of vacuum-filling nitrogen cycle, react in 90℃ oil bath for 10h, then place in liquid nitrogen to cool for 30s . The reaction product was precipitated with n-hexane, dialyzed for 3 days, and freeze-dried to obtain a yellow oil, which was a temperature-sensitive random polymer P(DEGMA-co-OVNGlu) with a molar ratio of OVNGlu and DEGMA of 1:4. The synthesis method of temperature-sensitive random polymers with OVNGlu and DEGMA molar ratios of 1:6 and 1:8 is the same as above, DEGMA is 0.3561g and 0.3685g, OVNGlu is 0.1029g and 0.0808g, respectively, figure 2 It is the synthetic route of P(DEGMA-co-OVNGlu).

[0045] The above-mentioned three kinds of random polymers with different ratios, thermosensitive monomer DEGMA and sugar ester monomer OVNGlu were tested...

Embodiment 2

[0049] Dissolve the 1:4 random polymer in Example 1 in the HEPES buffer to make polymers with concentrations of 0.005mg / mL, 0.05mg / mL, 0.1mg / mL, 0.3mg / mL, and 0.5mg / mL solution with a volume of 10 mL. The obtained series of solutions are filtered with a 0.45 μm aqueous phase filter membrane, and then added to a clean sample test bottle. First, test (the test angle is 90°, and the test time for each pair of interactions is 3min) of different concentrations of no The particle size of the regular polymer in HEPES buffer (D h ) and polydispersity (PDI), such as Image 6 shown. The results show that: for 37℃ nonlinear (T>LCST) random polymer, with the increase of polymer concentration, D h Gradually increases, PDI gradually decreases. When the polymer concentration is 0.005 mg / mL, the particle size of the polymer is 4.7nm, because the dust removal of the high polymer solution and its solvent is very important when testing light scattering, so the dust particles have a large eff...

Embodiment 3

[0051] (1) Prepare 0.4mg / mL P(DEGMA-co-OVNGlu): pipette 12mL of prepared 1mg / mL (20mL) P(DEGMA-co-OVNGlu) into a 50mL centrifuge tube, and then add 18 mL of the prepared HEPES buffer solution (pH=7.4), the obtained solution is 0.4 mg / mL P(DEGMA-co-OVNGlu) solution (30 mL).

[0052] (2) Prepare 0.4mg / mL Con A: Pipette 12mL of the prepared 1mg / mL (20mL) Con A into a 50mL centrifuge tube, and then add 18mL of the prepared HEPES buffer solution (pH = 7.4), the obtained solution is 0.4mg / mL Con A solution (30mL).

[0053] (3) Prepare 0.4mg / mL BSA: pipette 12mL of the prepared 1mg / mL (20mL) BSA into a 50mL centrifuge tube, and then add 18mL of the prepared HEPES buffer solution (pH=7.4) , the resulting solution was 0.4 mg / mL BSA solution (30 mL).

[0054] (4) Configure 0.4mg / mL RCA 120 : The prepared 1mg / mL (20mL) RCA 120 Pipette 12 mL into a 50 mL centrifuge tube, and then add 18 mL of the prepared HEPES buffer solution (pH=7.4), the resulting solution is 0.4 mg / mL RCA 120 sol...

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Abstract

The present invention relates to a method for testing the interaction of a temperature-sensitive random polymer with agglutinin. The method comprises the steps of: preparing a temperature-sensitive random polymer P (DEGMA-co-OVNGlu) solution; preparing an agglutinin protein Con A solution, an agglutinin protein BSA solution and an agglutinin protein RCA120 solution; UV testing the Con A solution,the BSA solution and the RCA120 solution; UV testing the mutual combination of the polymer with Con A, BSA, RCA120; subjecting different concentrations of polymer solutions to a dynamic light scattering test; and adding the agglutinin solution to the polymer solution and then performing a dynamic light scattering test. Compared with other test methods (surface plasmon resonance and isothermal titration calorimetry), the method is easy to operate, low in cost, and fast in test analysis.

Description

technical field [0001] The invention belongs to the field of measuring methods for the interaction between high molecular polymers and biomacromolecules, in particular to a measuring method for the interaction between temperature-sensitive random polymers and lectins. Background technique [0002] As an essential component in living organisms, sugar is one of the most abundant biomolecules in nature. Glycoproteins play important roles in the process of biological evolution, such as energy storage and metabolic intermediates, involved in signal transduction, immune response, cell growth regulation, cell-cell adhesion, cell differentiation and glycoprotein recognition. Among them, the recognition of sugar-protein plays a very important role in the process of biological evolution, but the molecular recognition ability of monovalent sugar is usually weak, and the interaction of sugar can be expanded by multivalent conjugation, which is usually called "sugar cluster". effect". ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/38G01N21/33G01N21/51
CPCG01N1/38G01N21/33G01N21/51
Inventor 权静郑永利张仪娜蔡豪
Owner DONGHUA UNIV
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