A method for detecting β-galactosidase based on surface enhanced Raman spectroscopy

By constructing Au NSs two-dimensional arrays and forming sandwich structures, combined with surface enhancement Raman technology, the existing β-galactosidase detection methods have solved the problems of large error, long time, low accuracy and high cost, and a rapid detection method with good sensitivity and specificity have been achieved.

CN115326779BActive Publication Date: 2025-04-01FUJIAN NORMAL UNIV
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Patent Information

Application Number
CN202210981029.7
Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Filing Date
2022-08-16
Publication Date
2025-04-01
Estimated Expiration
2042-08-16

AI Technical Summary

Technical Problem

The existing β-galactosidase detection methods have problems such as large errors, long staining time, low accuracy and high cost, and lack national and group standards.

Method used

Using a surface enhancement Raman technology (SERS)-based method, a two-dimensional array of Au NSs was constructed and attached to β-lactose after modification by PMBA, and connected to PATP-modified Au NSs to form a sandwich structure. The vitality of SA-β-gal was detected by laser Raman spectrometer.

Benefits of technology

It has achieved a more economical, faster and lower detection limit detection method, with high detection sensitivity, good specificity, wide application range, and can complete the detection within 1 hour, reducing operation difficulty and cost.

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Abstract

The present invention develops a method for detecting β-galactosidase in serum based on surface enhanced Raman spectroscopy technology. β-galactosidase is a marker enzyme for identifying senescent cells. The technical scheme includes: 1) preparing nano-Venus, 2) preparing a two-dimensional array of monolayer, self-assembled nano-Venus and labeled molecules of para-mercaptobenzoic acid, 3) preparing a label, and 4) combining the two-dimensional array with the label through lactose to form a functionalized gold plate as a detection platform. When β-galactosidase acts on the substrate, the two-dimensional array can be separated from the label by decomposing the β-galactosidic bond in lactose, and the data spectrum is collected by means of a laser Raman spectrometer, and the new signal 1437cm generated by PATP on the label when irradiated with laser ‑1 As the marker peak, the ultrasensitive detection of serum β-galactosidase content was achieved. When detecting the activity of the enzyme, the method ‑3 ‑0.5U·mL ‑1 and 0.5-100 U·mL ‑1 The detection limit can reach 4.79×10 ‑5 U·mL ‑1 .
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