Search

CN-116609310-B - CEA detection-based two-dimensional material biosensor and preparation method and application thereof

CN116609310BCN 116609310 BCN116609310 BCN 116609310BCN-116609310-B

Abstract

The invention relates to a two-dimensional material biosensor based on CEA detection, and a preparation method and application thereof, belonging to the technical field of two-dimensional material biosensing. According to the CEA detection-based two-dimensional material biosensor, a single-layer WS 2 crystal is grown on a Si/SiO 2 substrate by a chemical vapor deposition method, transferred on an ITO substrate by a wet transfer method, then the single-layer WS 2 crystal is treated by bis (trifluoromethane) sulfonyl imide, and an Anti-CEA probe protein is fixed by surface interface chemical modification, so that the biosensor is obtained. The single-layer WS 2 crystal prepared by the invention is easy to grow, has uniform fluorescence and better performance, and the biosensor has the characteristics of simple process, high sensitivity, low detection limit, no labeling of biomolecules and the like when used for detecting carcinoembryonic antigen.

Inventors

  • Xue tianyu
  • LIU ZHONGYUAN
  • LIU ZIQIAN
  • KONG JING
  • XIANG JIANYONG
  • NIE ANMIN
  • WEN FUSHENG
  • Mou Congpu
  • WANG BOLI
  • ZHAI KUN

Assignees

  • 燕山大学

Dates

Publication Date
20260508
Application Date
20230525

Claims (10)

  1. 1. A two-dimensional material biosensor based on CEA detection is characterized in that a single-layer WS 2 crystal is grown on a Si/SiO 2 substrate by a chemical vapor deposition method, transferred on an ITO substrate by a wet transfer method, then the single-layer WS 2 crystal is treated by bis (trifluoromethane) sulfonyl imide, and an Anti-CEA probe protein is fixed by surface interface chemical modification, so that the biosensor is obtained.
  2. 2. The method for preparing the two-dimensional material biosensor based on CEA detection as claimed in claim 1, comprising the following steps: (1) Transferring a single-layer WS 2 crystal grown on a Si/SiO 2 substrate by a chemical vapor deposition method onto the ITO substrate, removing the glue on the single-layer WS 2 crystal, placing the single-layer WS 2 crystal into a bis (trifluoromethane) sulfonyl imide solution, placing the single-layer WS 2 crystal into a hot plate for soaking, taking out the single-layer WS 2 crystal, drying the single-layer WS 2 crystal by nitrogen, and annealing the single-layer WS 2 crystal under the hot plate; (2) Soaking with 11-mercaptoundecanoic acid, soaking with carbodiimide hydrochloride and N-hydroxysuccinimide, and modifying the single-layer WS 2 crystal with Anti-CEA to obtain the biosensor.
  3. 3. The preparation method of claim 2, wherein the concentration of the bis (trifluoromethane) sulfonyl imide solution in the step (1) is 0.2 mg/mL, the soaking condition on the hot plate is that the temperature is 100-120 ℃ and the time is 10-12 min, and the annealing condition under the hot plate is that the temperature is 100-120 ℃ and the time is 5-6 min.
  4. 4. The preparation method according to claim 2, wherein the specific method for removing the glue on the single-layer WS 2 crystal in the step (1) is to soak the single-layer WS 2 crystal on the ITO substrate in acetone.
  5. 5. The method according to claim 4, wherein the soaking condition is that the temperature is 65-70 ℃ and the time is 15-20 min.
  6. 6. The preparation method of claim 2, wherein in the step (2), the concentration of the 11-mercaptoundecanoic acid is 50mM, the soaking time of the 11-mercaptoundecanoic acid is 24 hours, and the soaking treatment time of the carbodiimide hydrochloride and the N-hydroxysuccinimide is 4-6 hours.
  7. 7. The process according to claim 2, wherein the concentration ratio of the carbodiimide hydrochloride to the N-hydroxysuccinimide in the step (2) is 4:1, preferably the concentration of the carbodiimide hydrochloride is 400 mM/L and the concentration of the N-hydroxysuccinimide is 100 mM/L.
  8. 8. The method of claim 2, wherein the Anti-CEA modification in step (2) is performed for a time period of 12 h.
  9. 9. A kit for detecting carcinoembryonic antigen concentration is characterized in that, comprising the biosensor of claim 1.
  10. 10. The use of a CEA detection-based two-dimensional material biosensor of claim 1 in the preparation of a kit for detecting carcinoembryonic antigen concentration.

Description

CEA detection-based two-dimensional material biosensor and preparation method and application thereof Technical Field The invention belongs to the technical field of two-dimensional material biosensing, and particularly relates to a method for detecting carcinoembryonic antigen by using a biosensor. Background Carcinoembryonic antigen is an acidic glycoprotein with human embryo antigen characteristics, exists on the surface of cancer cells differentiated from endoderm cells, and is a structural protein of cell membranes. Formed in the cytoplasm, secreted out of the cell through the cell membrane and then into the surrounding body fluids. Thus, it can be detected from various body fluids and excretions such as serum, cerebrospinal fluid, milk, gastric juice, hydrothorax and ascites, urine, feces, etc. CEA is a major embryonic antigen isolated from adenocarcinoma and increases in body tissue in proportion to the number of cancer cells, and if the CEA content in the body increases, the probability of suffering from adenocarcinoma increases. CEA is used as a tumor marker, the expression level of which can accurately reflect the number, proliferation and extinction of cancer cells, and the content of CEA in healthy population is extremely low. When a patient develops tumor cells, the tumor marker content in the body can be abnormally increased in a short time, and can be used for assessing disease progression. CEA elevation is commonly found in colorectal cancer, pancreatic cancer, gastric cancer, breast cancer, liver cancer, lung cancer, ovarian cancer, urinary system tumors, and the like. Single-layer WS 2 crystals have many unique physical and chemical properties and have received extensive attention and research from the academic scientific community. The single-layer WS 2 crystal has the characteristics of wider controllable band gap, stronger photoluminescence efficiency, excellent carrier mobility and the like, and has a wider application prospect in the fields of biosensing and the like. The current methods for preparing single-layer WS 2 crystals mainly comprise a mechanical stripping method, a liquid phase stripping method and a chemical vapor deposition method. The band gap of the single-layer WS 2 crystal is about 1.97eV, and the single-layer WS 2 crystal has stronger photoluminescence efficiency and light absorption performance. However, due to structural defects such as sulfur atom vacancies and the like of the single-layer WS 2 crystal, the problems of poor fluorescence and chemical stability and the like of the single-layer WS 2 crystal can be caused. In the prior art, document "ADNAzyme-catalyzedlabel-freeaptasensorbasedonmultifunctional dendrimer-likeDNAassemblyforsensitivedetectionofcarcinoembryonicantigen" discloses a method for detecting CEA based on a multifunctional dendrimer-like DNA nano-assembly that creates a simple, label-free and antibody-free aptamer sensor. However, the modification process of the document is complex, the operation process is more complex, the detection limit is higher, and the industrialization is not facilitated. Document "Magneticantifoulingmaterialbasedratiometricelectrochemicalbiosensorforthe accuratedetectionofCEAinclinicalserum" discloses a novel electrochemical biosensor based on a multifunctional nanocomposite. However, the sample synthesis process of the document is complex, the time is prolonged, the cost is high, the detection limit is high, and the industrialization is not facilitated. In summary, the main disadvantages of the current CEA biosensor are that, firstly, the detection limit is higher and the detection sensitivity is lower. Secondly, the substrate sample synthesis process is relatively complicated. Thirdly, the preparation process is complex, has high requirements on equipment and high cost, and is not suitable for mass production. Disclosure of Invention Aiming at the problems in the prior art, the invention aims to design and provide a CEA biosensor with low detection limit, high detection sensitivity and rapid and efficient detection and a preparation method thereof. The single-layer WS 2 crystal prepared by the invention is easy to grow, has uniform fluorescence and better performance, and the biosensor has the characteristics of simple process, high sensitivity, low detection limit, no labeling of biomolecules and the like when used for detecting carcinoembryonic antigen. Binding of the monolayer WS 2 crystals to carcinoembryonic antigen resulted in a change in the fluorescence signal of the monolayer WS 2 crystals, thereby detecting the concentration of carcinoembryonic antigen. The CVD method is mainly used in the invention to grow high quality single-layer WS 2 crystals. The single-layer WS 2 crystal is treated by using the TFSI to repair or eliminate the problem of poor fluorescence and chemical stability caused by structural defects such as sulfur atom vacancy and the like, thereby improving the fluorescence and chemical stability