The Development Of The Fluorescence Immunochromatographic Strip

In this study we chose salbutamol as target detection,selected quantum dots and Upconversion Fluorescence the two kinds of fluorescent material as markers,to study the two fluorescence immunochromatographic strip analysis method,and applied to the analysis of salbutamol residues in food of animal origin.Purified the albutamol antibody through Protein A-Sepharose 4B,obtained the antibody concentration is 2.77 mg mL^-1.Through the method of activated ester to realize the conjugation of antibody and quantum dots?CdTe-COOH-PEG?,selected quantum dots:antibody:EDC = 1:10:3000?molar ratio?as the best reaction ratio,in order to obtain purified antibody quantum dot compound,after the reaction used size exclusion column to purify the product.Optimized the reaction liquid pH is 8.3,connection time is 3 h,the loading solution is phosphate buffer,antibody quantum dot compound adding amount is 2 ?L,working liquid adding amount is 10 ?L,the dilute time of the second antibody and the coating antigen is 100 times and 18 times,selected the HF135 NC membrane,used 0.5%BSA to handle the sample pad.Under the optimal condition to assemble the strip,using the method of wet process,the method detection limit is 2.0 ?g L^-1.Chose pork,beefinutton,pork liver,pork kidney,chicken,six kinds of actual samples to do the sanmple detection,the sample detection limit is 10?g kg^-1.Synthesized UCNPs by using Hydrothermal method,and synthesized particle size uniform NaYF₄:Er³⁺,Yb³⁺?OA-UCNPs?,used polyacrylic acid?PAA?by ligand exchange method at high temperature modified OA-UCNPs to water-soluble PAA-UCNPs.Through the method of activated ester to realize the conjugation of antibody and PAA-UCNPs.Optimized the best amount Ab to conjugated the UCNPs is 0.5 mg/5 mg,the dilute time of the second antibody and the coating antigen is 10 times and 18 times,the antibody-UCNPs compound adding amount is 10 ?L,the loading solution is phosphate buffer,working liquid adding amount is 10 ?L.Under the optimal condition to assemble the strip,using the method of wet process,the method detection limit is 5.0 ?g L^-1.Chose pork,beef,mutton,pork liver,pork kidney,five kinds of actual samples to do the sample detection,the sample detection limit is 25 ?g kg^-1.The study developed two fluorescence immunochromatographic strip analysis method to detect salbutamol.The methods have the advantage of strong specificity,high sensitivity,small background interference,short time-consuming and easy to judge the result,is suitable for salbutamol residues detection in the meat.