A homogeneous method troponin chemiluminescent detection reagent and preparation method

Abstract

The invention discloses a homogeneous method troponin chemiluminescence detection reagent and a preparation method, comprising the steps of using nano-gold particles, adding a water-soluble photosensitizer to the nano-gold particles to obtain a nano-gold-photosensitizer complex, and then adding nano The gold-photosensitizer complex is then coupled with the troponin I monoclonal antibody, and the troponin I monoclonal antibody is electrostatically adsorbed under its isoelectric point environment through the charge of the gold nanoparticles. , so that troponin I monoclonal antibody is coupled to the surface of gold nanoparticles. A homogeneous troponin chemiluminescence detection reagent obtained by the preparation method of the present invention improves the utilization rate of the photosensitizer, simplifies the production process, reduces production costs, and uses the sandwich method detection principle to detect the detection substance during use. The effective light dose of the irradiation is increased, more photosensitizers are activated and a stronger photosensitizing effect is produced, and the difference between the excitation light wavelength and the emission wavelength is increased, thereby improving the recognition degree of detection.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a homogeneous troponin chemiluminescent detection reagent and a preparation method. Background technique [0002] Acute myocardial infarction (AMI) is an emergency and critical disease of the cardiovascular system, with high morbidity, poor prognosis and high mortality. For patients with myocardial infarction without typical chest pain and insignificant ECG changes, it is difficult to accurately diagnose only by ECG, echocardiography, and cardiac MRI. Most of the traditional detection items are myocardial enzyme spectrum, but there are deficiencies such as late rise of enzyme activity, poor specificity and short duration. Cardiac troponin is the only contractile protein present in the myocardium and is highly sensitive and specific to myocardial necrosis. [0003] cTnI is one of the three subunits (cTnI, cTnC and cTnT) of cardiac troponin (cTn), which is a regulatory protein of mus...

AI Technical Summary

Problems solved by technology

[0006] For the detection method of troponin I, the existing light-induced chemiluminescence method needs to use non-water-soluble photosensitizers (porphyrin, chlorophyll, phthalocyanine and their derivatives, etc.), and singlet oxygen quantum production of unmodified photosensitizers The rate is low, which affects the detection performance; and the derivatized photosensitizer, the chemical synthesis is difficult and the process is complicated

[0007] The photosensitizer used in the existing photoluminescence system needs to be embedded in polystyrene microspheres in the organic phase, which needs to be purified through complicated centrifugation, ultrafiltration, etc., and needs to be restored to the aqueous phase matrix. It is easy to produce loss during the process of burying and purification, which reduces the photosensitive effect

[0008] Existing light-induced chemiluminescence technology has little difference between the excitation wavelength and the detection wavelength, and is susceptible to interference, and the fluorescence background is greatly affected

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