|
Summary of various domestic and international papers in the microfluidic chip enzymatic reaction analysis system , based on a variety of microfluidic chips compared enzyme reaction analysis system characteristics, selected styrene monolith immobilized enzyme microarray analysis system , followed by the analysis system has been applied research . First, the functional monomer styrene , divinyl benzene crosslinking agent ethanol pore azobisisobutyronitrile (AIBN) as initiator , UV initiator , preparation of the whole inverted polystyrene column material ; to meet their requirements for enzyme immobilization , the use of sulfuric acid to obtain a sulfonated cation- modified polystyrene monolith material . By scanning electron microscopy , characterized by means of infrared spectroscopy of the structure and morphology of the material properties and the flow rate of the solution through the column is less than 1.5mL/min , the measured length of 10mm, a diameter of 1mm overall modified polystyrene column samples of L-Tyr maximum adsorption of about 3.4mg, column capacity about 0.43mg/mm3, and has good mechanical stability. GOD enzyme fixed on a polystyrene dielectric material , the specifications for the long 20mm, 1mm diameter polystyrene monolithic column , with UV detection benzoquinone measured by the method , when used in the free enzyme is 10U , GOD immobilized enzyme about 7U,, enzyme immobilization efficiency of 70%. In the microfluidic chip pipeline to achieve a monolithic column situ polymerization of polystyrene and sulfonated modified in modified polystyrene monolithic column immobilized HRP enzyme to determine the HRP enzyme in the enzyme concentration 5μg / mL and speed 0.2mL/min through the column , the fixed effect is good , in order to build a styrene monolithic column immobilized enzyme microreactor chip analysis systems . The microreactor chip analysis systems combined with chemiluminescence detection methods , the establishment of a whole column immobilized enzyme microreactor chips - chemiluminescence detection method. Chemiluminescence optimization experiments found that the sample flow rate through film 0.2mL/min, luminescent reagent and sample volume ratio of 1:1 , you can achieve the best chemiluminescence detection. With ascorbic acid (Vc) and the method of levodopa two samples was verified , to obtain a good detection. The analysis method , by fixing different enzymes or antibodies , can be used for a corresponding enzyme substrate or antigen detection with broad applicability.
|