Preparation Of Carboxylated Polystyrene Fluorescent Nanoparticles And Its Application In Homogeneous Luminescence Immunity

Objective:The light-excited chemiluminescence homogeneous immunoassay technology for singlet oxygen channels using polystyrene fluorescent nanoparticles is the third-generation homogeneous immunoassay technology.The advantage of this technology is that the range of substances to be measured is wide,the speed of detection is fast,stable and it has higher sensitivity.During the detection process,The introduction of fluorescent tags is not required,which prevents steric hindrance from affecting the mutual binding of biomolecules.It can also be used to detect crude biological extracts without affecting the reading effect.However,the current domestic research on this aspect is not enough,because the photoexcitation chemiluminescence nanoparticles required for detection mainly rely on foreign,and the materials are expensive.As a result,it cannot be widely used.Therefore,Our study is focus on the preparation of photo-excited chemiluminescence nanoparticle,and to establish”Latex aggregation-triggered Amplified luminescent assay”detection system.Methods:(1)The emulsion polymerization method was used to study the preparation of polystyrene nanoparticles.The study used SDBS or SDS as emulsifiers,and acrylic acid was used to modify the surface of the nanoparticles.(2)Two kinds of fluorescent complexes were prepared with metal europium.The fluorescent substance with the largest fluorescence intensity was selected to react with the nanoparticles to synthesize fluorescent nanoparticles using the method of high temperature swelling adsorption method.The effect of the second fluorescent substance BPEA on the fluorescence intensity of fluorescent nanoparticles was also investigated.Then use the same method to synthesize photosensitive nanoparticles with materials containing phthalocyanine.Also use fluorescent complex,BPEA,thioxene to react with nanoparticles to synthesize luminescent nanoparticles.(3)"Soaking"the photosensitive nanoparticles into the luminescent solution,and"Soaking"the photosensitive nanoparticles into the photosensitive solution for testing,and verifying whether the two hands have photochemical chemiluminescence detection properties.Then Hcg detection and?₂-MG detection have been verified that the photo-induced chemiluminescence nanoparticles can be applied in actual detection.Results:(1)By adjusting the amount of SDS and SDBS,the carboxylated polystyrene nanoparticles with controllable particle size in the range of 70-250 nm were successfully prepared.And nanoparticles with different carboxy density were studied by changing the amount of acrylic acid added.The prepared nanoparticles have a regular spherical shape,uniform particle size,and good dispersibility.(2)Two kinds of fluorescent complexes were successfully synthesized.It can be seen from the comparison that Eu(TTA)₃phen has stronger fluorescence intensity,and its brightness is more obvious under UV light irradiation.Using Eu(TTA)₃phen as the fluorescent substance,carboxylated polystyrene fluorescent nanoparticles were synthesized under optimized conditions.After the addition of BPEA,the intensity of the fluorescent nanoparticles was stronger.Based on this method,photosensitive nanoparticles and luminescent nanoparticles with special properties are successfully prepared.(3)Experiments shown that the prepared photosensitive nanoparticles and luminescent nanoparticles have the special properties of photo-induced chemiluminescence detection.Different detection experiments and conditions have been tested.The obvious light-induced chemiluminescence signal value shown that the light-induced chemiluminescence particles can be successfully applied to immunoassay.Conclusion:In this project,carboxylated polystyrene fluorescent nanoparticles were successfully prepared,and the particle size was controlled and the carboxyl density was adjustable.After the successful synthesis of fluorescent complexes,a method for preparing carboxylated polystyrene fluorescent nanoparticles was developed.On this basis,photosensitive nanoparticles and luminescent nanoparticles suitable for photo-excitation chemiluminescence immunodetection systems were prepared.And Photosensitive nanoparticles and luminescent nanoparticles were successfully used in detection.So Latex aggregation-triggered Amplified luminescent assay was initially established.This method lays the foundation for further research in this area.