Search

CN-121995047-A - Dual ligand peptide/UIO-66-NH2Preparation method of @ AYG fluorescent probe

CN121995047ACN 121995047 ACN121995047 ACN 121995047ACN-121995047-A

Abstract

The application belongs to the technical field of biomedical detection, and provides a preparation method of a dual ligand peptide/UiO-66-NH 2 @ AYG fluorescent probe and application thereof in CA153 detection, the fluorescent probe has the advantages of high sensitivity, high specificity, good stability, low cost, simple operation and the like, can meet the accurate detection requirement of trace CA153, and is particularly suitable for basic medical institutions and large-scale clinical screening.

Inventors

  • WEI ZEHUI
  • GE SHUXIAN
  • SHEN XIN
  • MOU LINA
  • TIAN MEI
  • SUN TAO

Assignees

  • 江苏海洋大学

Dates

Publication Date
20260508
Application Date
20251230

Claims (10)

  1. 1. A method for preparing a dual ligand peptide/UiO-66-NH 2 @ AYG fluorescent probe, the method comprising: Step 1, mixing UiO-66-NH 2 , a dual ligand peptide, a cross-linking agent and a reaction solvent, and performing constant temperature reaction to obtain dual ligand peptide/UiO-66-NH 2 composite nano particles, wherein the dual ligand peptide comprises SEQ ID NO.1: GTTFSNYW and SEQ ID NO.2: MHYLEYPF, and the cross-linking agent comprises o-phthalaldehyde (OPA); And 2, mixing and incubating the dual-ligand peptide/UiO-66-NH 2 composite nano particles with a fluorescent signal molecule Acridine Yellow G (AYG) at room temperature to obtain the dual-ligand peptide/UiO-66-NH 2 @ AYG fluorescent probe.
  2. 2. The preparation method of claim 1, wherein the step 1 comprises mixing UiO-66-NH 2 , GTTFSNYW, MHYLEYPF and phthalic aldehyde in proportion, performing ultrasonic dispersion for 1-3 min, performing nitrogen purging for 4-6 min, performing a water bath constant temperature reaction at 60-70 ℃ under a sealed condition, wherein the reaction solvent is one of methanol, N-butanol, dimethyl sulfoxide and N, N-dimethylformamide, preferably dimethyl sulfoxide, and the reaction time is 6-30 h, preferably 18-24 h.
  3. 3. The preparation method according to any one of claims 1 to 2, wherein the step 2 comprises the steps of mixing the dual ligand peptide/UiO-66-NH 2 composite nano-particles with a concentration of 50-500 μg/mL, acridine yellow G with a concentration of 5-15 μg/mL at room temperature for 20-40 min, and the reaction solvent is methanol.
  4. 4. A method of preparation according to any one of claims 1 to 3 wherein the molar ratio of GTTFSNYW to MHYLEYPF in the dual ligand peptide is 1:1.
  5. 5. The method according to any one of claims 1 to 4, wherein the molar ratio of GTTFSNYW, MHYLEYPF to phthalic aldehyde is 1:1 (1 to 24), preferably 1:1 (12 to 24), more preferably 1:1:18.
  6. 6. The method of any one of claims 1 to 5, wherein the molar ratio of UiO-66-NH 2 , GTTFSNYW, MHYLEYPF, o-phthalaldehyde is 1:1:1 (1-24), preferably 1:1:1 (12-24), more preferably 1:1:1:18.
  7. 7. The process according to any one of claim 1 to 6, wherein, Mixing UiO-66-NH 2 , GTTFSNYW, MHYLEYPF and phthalic aldehyde in proportion, performing ultrasonic dispersion for 1-3 min, then purging nitrogen for 4-6 min, performing water bath constant temperature reaction at 60-70 ℃ under a sealing condition, wherein the reaction solvent is dimethyl sulfoxide, the reaction time is 24h, and the feeding mole ratio of the UiO-66-NH 2 , GTTFSNYW, MHYLEYPF and the phthalic aldehyde is 1:1:1:18; The step 2 comprises the steps of reacting the dual-ligand peptide/UiO-66-NH 2 composite nano-particles with a fluorescent signal molecule Acridine Yellow G (AYG) to obtain a dual-ligand peptide/UiO-66-NH 2 @ AYG fluorescent probe, wherein the concentration of the dual-ligand peptide/UiO-66-NH 2 composite nano-particles is 500 mug/mL, the concentration of acridine yellow G is 15 mug/mL, the reaction time is 20-40 min, and the reaction solvent is methanol.
  8. 8. A dual ligand peptide/UiO-66-NH 2 @ AYG fluorescent probe prepared according to the method of any one of claims 1 to 7.
  9. 9. Use of a dual ligand peptide/UiO-66-NH 2 @ AYG fluorescent probe prepared by the method of any one of claims 1 to 7 in CA153 detection.
  10. 10. The use according to claim 9, wherein the test object comprises a mouse serum labeled sample or a clinical cancer patient serum sample, and the fluorescent probe and the sample are incubated for 10-40 min in a dark place under vibration, and the adsorption equilibrium time is 10-30 min.

Description

Preparation method of dual ligand peptide/UIO-66-NH 2 @ AYG fluorescent probe Technical Field The application relates to the technical field of biomedical detection, in particular to a preparation method of a double ligand peptide/UIO-66-NH 2 @ AYG fluorescent probe and application thereof in ultra-sensitive detection of saccharide antigen 153 (CA 153). Background Saccharide antigen 153 (CA 153) is a specific epitope of a mucinous glycoprotein encoded by MUC-1 gene, is used as a detection index of recurrence of II/III stage breast cancer, and is one of core serum tumor markers most closely related to breast cancer in clinic. A large number of clinical researches prove that the concentration of CA153 in serum of a breast cancer patient is obviously higher than that of a benign breast disease patient, the concentration of CA153 in serum of an early breast cancer patient is extremely low (only 20% -40% of the rise occurs), and the dynamic change of the concentration can provide a key basis for early diagnosis, disease progress evaluation, curative effect monitoring and recurrence early warning of breast cancer. In addition, CA153 can also be used as an auxiliary diagnosis marker for other malignant tumors such as lung cancer, ovarian cancer, cervical cancer and the like, and provides an important reference for clinical screening and differential diagnosis of related diseases. At present, the clinical routine detection CA153 mainly adopts an enzyme-linked immunosorbent assay (ELISA), and other detection technologies such as Surface Enhanced Raman Scattering (SERS), a microfluidic biosensor, an electrochemiluminescence immunoassay and the like are reported in related researches. Although the technologies have certain detection performance, the defects of complex operation flow and high requirements on professional skills of experimental equipment and operators generally exist, and particularly, the technologies have practical application limitations in special occasions such as medical resource deficiency scenes, large-scale community screening, invasive sampling difficulties and the like, and are difficult to popularize widely. Therefore, the development of the fluorescent probe based on the UiO-66-NH 2 and the dual ligand peptide is expected to solve the defects of the existing CA153 detection technology, realize the detection requirements of ultrasensitivity, low cost and convenient operation, and have important clinical application value. Disclosure of Invention The application aims to provide a preparation method of a dual ligand peptide/UIO-66-NH 2 @ AYG fluorescent probe and application thereof in CA153 detection, and the fluorescent probe has the advantages of high sensitivity, high specificity, good stability, low cost, simplicity and convenience in operation and the like, can meet the accurate detection requirement of trace CA153, and is particularly suitable for basic medical institutions and large-scale clinical screening. The technical scheme is that in the first aspect, the application provides a preparation method of a dual ligand peptide/UIO-66-NH 2 @ AYG fluorescent probe, which is characterized by comprising the following steps of: step 1, mixing UiO-66-NH 2, a dual ligand peptide, a cross-linking agent and a reaction solvent, and performing constant temperature reaction to obtain dual ligand peptide/UiO-66-NH 2 composite nano particles, wherein the dual ligand peptide comprises SEQ ID NO.1: GTTFSNYW and SEQ ID NO.2: MHYLEYPF, and the cross-linking agent comprises o-phthalaldehyde (OPA); And 2, mixing and incubating the dual-ligand peptide/UiO-66-NH 2 composite nano particles with a fluorescent signal molecule Acridine Yellow G (AYG) at room temperature to obtain the dual-ligand peptide/UiO-66-NH 2 @ AYG fluorescent probe. In some embodiments, the step 1 comprises mixing UiO-66-NH 2, GTTFSNYW, MHYLEYPF and phthalic dicarboxaldehyde in proportion, performing ultrasonic dispersion for 1-3 min, then purging nitrogen for 4-6 min, and performing a water bath constant temperature reaction at 60-70 ℃ under a sealing condition. In some embodiments, the reaction solvent is one of methanol, N-butanol, dimethylsulfoxide, N dimethylformamide, preferably dimethylsulfoxide. In some embodiments, the reaction solvent is one of methanol, N-butanol, dimethyl sulfoxide, N dimethylformamide in an amount of 20mL (corresponding to 0.01mmol of UiO-66-NH 2). In some embodiments, the reaction time is 6 to 30 hours, preferably 18 to 24 hours, more preferably 24 hours. In some embodiments, the step 2 comprises the steps of mixing and incubating the double ligand peptide/UIO-66-NH 2 composite nano-particle with the concentration of 50-500 mug/mL and the acridine yellow G with the concentration of 5-15 mug/mL at room temperature for 20-40 min, wherein the reaction solvent is methanol. In some embodiments, the molar ratio of GTTFSNYW to MHYLEYPF in the dual ligand peptide is 1:1. In some embodiments, the GTTFSNYW, MHYLEYPF,