Sensor detection kit based on 96-well plate immobilized molecular motor and method for in-vitro detection of P24 antigen

Abstract

The invention discloses a sensor detection kit based on a 96-well plate immobilized molecular motor and a method for P24 antigen in vitro detection.The kit is composed of a molecular motor sensor detection plate, and the detection plate is formed by immobilizing an F0F1-ATP enzyme molecular motor on a 96-well ELISA plate; meanwhile, a detection probe (P24 antibody) is combined to form a molecular motor biosensor for capturing a target. The detection method mainly comprises the following steps: (1) placing a sample in the detection plate, and capturing a target object by the sensor; (2) adding an ATP (adenosine triphosphate) synthesis solution containing ADP (adenosine triphosphate), and reacting And (3) adding an ATP detection reagent, quickly and uniformly mixing, and starting a chemical light emission detector to measure the luminous intensity. According to the present invention, the stability and the uniformity of the 96-well plate immobilized molecular motor, the mutual conversion characteristic of the kinetic energy and the chemical energy of the molecular motor, and the high sensitivity and the rapid performance of the chemiluminescence are utilized to detect the antigen, and the porosity of the 96-well plate enables the rapid and stable detection to be achieved while the high throughput screening requirement can be met.

Description

technical field [0001] The invention belongs to the field of in vitro diagnostic reagents, in particular to a sensor detection kit based on a 96-well plate immobilized molecular motor, and a method for in vitro detection of P24 antigen. Background technique [0002] Biomolecular motors are proteases that convert chemical energy into mechanical energy. Among them, ATP synthase, also known as F0F1-ATP synthase (F0F1-ATPase), is widely present in the mitochondrial inner membrane of animals and fungi, plant chloroplast thylakoid membranes and bacteria. The plasma membrane is the main site of ATP synthesis and is the smallest known rotary molecular motor. F0F1-ATP synthase can not only synthesize ATP using the transmembrane proton gradient and drive the rotor to rotate clockwise, but also hydrolyze ATP to transport protons and drive the rotor to rotate counterclockwise. Due to the special motion and rotation characteristics of F0F1-ATPase, it can be prepared as a molecular motor...

AI Technical Summary

Problems solved by technology

[0003] Traditional molecular motor biosensor detection kits often immobilize molecular motors on the surface of magnetic beads to construct biosensors, but this reconstruction method relies on the biotin-streptavidin-biological system to connect magnetic beads and molecular motors, which is easy Generate magnetic beads - ineffective by-products of magnetic beads

Because the magnetic beads are easy to settle, the batch-to-batch variation in the detection time is large, and the operation of the magnetic bead molecular motor biosensor is cumbersome and repeatable in large-scale detection.

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