Homogeneous method heart-type fatty acid binding protein chemiluminiscence detection reagent and preparation method thereof

Abstract

The invention discloses a homogeneous method heart-type fatty acid binding protein chemiluminiscence detection reagent and a preparation method thereof. The method comprises steps of adopting gold nanoparticles, adding a water-soluble photosensitizer into the gold nanoparticles, acquiring a nanogold-photosensitizer compound, coupling the nanogold-photosensitizer compound with a heart-type fatty acid binding protein monoclonal antibody, generating electrostatic adsorption between the two under the isoelectric point environment condition of the heart-type fatty acid binding protein monoclonal antibody through the charge property of the gold nanoparticles, so that the heart-type fatty acid binding protein monoclonal antibody is coupled to the surfaces of the gold nanoparticles. The homogeneous method heart-type fatty acid binding protein chemiluminiscence detection reagent is obtained according to the preparation method disclosed by the invention; the utilization rate of the photosensitizer is increased, the production process is simplified, the production cost is reduced, a sandwich method detection principle is adopted to detect a detected object in use, the effective light dose ofirradiation is increased, more photosensitizers are activated, a stronger photosensitive effect is generated, the difference between the excitation light wavelength and the emission wavelength is increased, and the detection identification degree is improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a homogeneous core fatty acid binding protein chemiluminescence detection reagent and a preparation method. Background technique [0002] Heart-type fatty acid-binding protein (h-FABP) is a novel small cytoplasmic protein that is abundant in the heart and is highly cardiac-specific (ie, mainly expressed in cardiac tissue). Following myocardial ischemic injury, h-FABP can be detected in the blood as early as 1 to 3 hours after the onset of chest pain, peaking at 6 to 8 hours and plasma levels returning to normal within 24 to 30 hours. The cardiac fatty acid-binding cytoplasmic protein consists of 132 amino acids with a molecular weight of 15 kDa. The h-FABP gene is located on chromosome I. It is one of the most abundant proteins in the heart. h-FABP binds two fatty acid molecules and participates in the transport of fatty acyl-CoA, which is active in the oxidation process to genera...

AI Technical Summary

Problems solved by technology

[0004] For the detection method of heart-type fatty acid binding protein, the existing light-induced chemiluminescence method needs to use non-water-soluble photosensitizers (porphyrin, chlorophyll, phthalocyanine and their derivatives, etc.), and the singlet oxygen quantum of unmodified photosensitizers The yield is low, which affects the detection performance; and the derivatized photosensitizer is difficult to synthesize and the process is complicated

[0005] 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

[0006] 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

Images