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Preparation method and chiral separation application of poly-dopamine/oxidized grapheme/BSA (Bovine Serum Albumin)

A polydopamine and graphene technology, which is applied in chemical instruments and methods, ion exchange, ion exchange regeneration, etc., can solve the problems of low column capacity compared to the column, limiting the application of OT-CEC, etc., and achieves low cost, high efficiency separation, Simple and fast effects

Inactive Publication Date: 2013-10-23
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the amount of stationary phase in OT-CEC is relatively small, making the comparison and column capacity low, which greatly limits the application of OT-CEC in separation

Method used

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  • Preparation method and chiral separation application of poly-dopamine/oxidized grapheme/BSA (Bovine Serum Albumin)
  • Preparation method and chiral separation application of poly-dopamine/oxidized grapheme/BSA (Bovine Serum Albumin)
  • Preparation method and chiral separation application of poly-dopamine/oxidized grapheme/BSA (Bovine Serum Albumin)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Fabrication of PDMS chip: Using the SU-8 positive mold (Boao Biological Co., Ltd.) as a template, make a typical cross-shaped PDMS microfluidic chip channel, such as figure 1 shown. The specific production process is as follows: Take a certain amount of PDMS monomer and curing agent according to 10:1 (mass ratio), mix evenly, degas, pour on the SU-8 template, and cure at 70 oC for 2 hours. After cooling, peel off the PDMS chip containing the cross-shaped channel from the template, cut it into the desired shape with a blade, and punch holes in the buffer pool, sample pool and sample waste pool with a puncher to form a diameter of 3 mm holes. At the same time, using the smooth glass plate as a template, follow the same steps to prepare a PDMS chip without microchannels as a cover slip. The PDMS chip containing the cross channel and the PDMS cover sheet without the channel were ultrasonically cleaned with secondary water, methanol, and secondary water for 10 minutes resp...

Embodiment 2

[0024] (1) Using the strong adhesion properties of PDA to immobilize GO in the separation channel of the PDMS microfluidic chip: mix a certain concentration of DA and GO in a Tris-HCl buffer solution with a pH of 8.5, and pump it into the PDMS microfluidic flow with a vacuum pump. In the separation channel of the control chip, the solution was continuously pumped for 5 minutes to cover the entire separation channel, and left to react at room temperature for 2 hours, and the PDA / GO functionalized PDMS microfluidic chip was obtained;

[0025] (2) Preparation of PDA / GO / BSA functionalized PDMS microchip: Wash the separation channel of the PDA / GO functionalized PDMS microfluidic chip prepared in the above steps with PBS buffer solution for 5 min, and then use a vacuum pump to inject 1 mg / mL The BSA solution was pumped into the separation channel, continuously pumped for 2 min, and allowed to stand at 4 °C for 4 h to obtain a PDA / GO / BSA functionalized PDMS microfluidic chip, which wa...

Embodiment 3

[0030] Application of PDA / GO / BSA functionalized PDMS microfluidic chip:

[0031] (1) An important application of microfluidic chips is the separation of analytes, Figure 5 For (A) 3.5 mM D-Tryptophan and 7.0 mM L-Tryptophan, (B) 3.5 mM D-Threonine and 7.0 mM L-Threonine, (C) 4 mM Gly-D-Phe and Electrochromatographic separation curves of 4 mM Gly-L-Phe on (a) PDMS chip and (b) PDA / GO / BSA modified PDMS chip. Experiments were carried out in 20 mM PBS buffer, pH 7.17, with an injection voltage of 800 V, a separation voltage of 1300 V, and a detection potential of +0.6 V. Depend on Figure 5 It can be seen that in the PDMS chip microchannel, chiral amino acids and dipeptides have only one peak, which cannot be effectively separated; however, in the PDA / GO / BSA modified PDMS chip microchannel, tryptophan, threonine and Enantiomers such as dipeptide were separated efficiently, with resolutions of 1.57, 1.76 and 1.74, respectively. The above results show that the PDA / GO / BSA modifi...

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Abstract

The invention relates to an immobilization method of bovine serum albumin taking poly-dopamine / oxidized graphene as a second-order reaction platform in a micro-fluidic chip channel and chiral separation application of the bovine serum albumin, and belongs to the technical field of micro-fluidic chips. The immobilization method comprises the following steps of: pumping a dopamine and oxidized graphene mixed solution into a micro-fluidic chip separation channel by using a vacuum pump, wherein the dopamine fixes the oxidized graphene to the surface of the micro-fluidic chip separation channel to form a poly-dopamine / oxidized graphene film in a self-polymerization process; combining the bovine serum albumin with the surface of poly-dopamine / oxidized graphene under the action of Pi-Pi, a hydrogen bond, hydrophobicity, and the like between the poly-dopamine / oxidized graphene and the bovine serum albumin to obtain a poly-dopamine / oxidized graphene / bovine serum albumin functionalized micro-fluidic chip channel. The bovine serum albumin has good stability, biocompatibility and hydrophilicity, can be used for enhancing the loading capacity of the bovine serum albumin in the micro-fluidic chip separation channel, keeping the biological activity of the bovine serum albumin and providing the universal platform for the high-efficiency separation of an amino acid antipode and a dipeptide antipode.

Description

technical field [0001] The invention relates to a method for realizing BSA immobilization in a PDMS microchip channel by using polydopamine / graphene oxide as a secondary reaction platform and its application in chiral separation. Background technique [0002] Chirality is a common phenomenon in nature, and chiral compounds are divided into D-type and L-type. In an achiral environment, the physical and chemical properties of the D- and L-isomers are similar. While in chiral environment, especially in biological environment, they exhibit different pharmacological properties and biological activities. Therefore, chiral separation is particularly important in the fields of pharmacy and biochemistry. Separation of chiral enantiomers is difficult due to their similar physical and chemical properties. Capillary electrochromatography (CEC) has both the high efficiency of capillary electrophoresis and the good selectivity of liquid chromatography. In the past few decades, CEC has ...

Claims

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Application Information

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IPC IPC(8): B01L3/00B01D15/08
Inventor 梁汝萍刘春鸣邱建丁
Owner NANCHANG UNIV
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