Mixed Protein Electrophoresis Titration Model And Leverage Princeple

Chip electrophoresis is an fundamental branch of capillary electrophoresis and a promising application tool.It is widely applied in analytical chemistry,life sciences,food testing,clinics,and other fields due to its integrated,cheap and easy to be commercialized,fast analysis speed,and portable characteristics.However,chip electrophoresis combined with Moving Reaction Boundary(MRB)was barely applied to the detection of mixed protein samples.Herein,we detect the ratio of proteins in mixed protein samples and real milk sample adultrated with soybean milk via chip platform integrating MRB.The specific contents are as follows:1.Design of chip and the construction of chip analysis system.We designed our chip with AutoCAD software and made it with electric processing technology.The size of the chip is 9.2 cm ? 4.5 cm with 30 units,which are uniformly distributed on its surface.Each unit consists of anode groove(3.5mm in diameter),cathode groove(3.5mm in diameter),and 6 parallel separation channels(0.2mm depth ? 0.2mm width ? 8mm length).In the Chip analysis system,chip acts as the carrier,and mercury lamp acts as the excitation light source.Then the excitation light goes through Olympus fluorescence microscope and finally irradiates detection channel.The sample in the channel is inspired by the excitation light and emits fluorescence.The emission fluorescence reflects to a CCD camera,and the data are collected by Toup View software.Ultimately a Meta Morph software processes and analyzes the data.Overall,the chip analysis system is composed of chip,mercury lamp,fluorescent microscope,Toup View software,and Meta Morph software.2.Retardation signal leverage theory in the titration of mixed protein samples.In the present work we address a simple,rapid and quantitative analytical method for detection of different proteins in biological samples.For this,we proposed the model of titration of mixed proteins(TMP)and its relevant leverage theory relied on the retardation signal of chip moving reaction boundary electrophoresis(MRBE).The leverage principle showed that the product of the first protein content and its absolute retardation signal is equal to that of the second protein content and its absolute one.3.Establishment of TMP model and the demonstration of the leverage principle.We chose BSA,Pepsin and protamine as model proteins.Every two model proteins were mixed with different proportion(1:1,1:2,1:3).Then the relevant experiments were conducted on the TMP-MRBE chip.The detection results based on leverage principle of TMP-MRBE agree with the actual protein ratio,which revealed that there was a leverage principle of retardation signal within the TMP of two pure proteins.4.Application of leverage theory in TMP-MRBE in real sample.The developed method was successfully applied to the determination of infant milk powder forged with soymilk powder via acid TMP-MRBE.The whole process of screening adulterated milk was from qualitative analysis via PAGE-MS to the quantitative determination by TMP-MRBE chip.Finally,we prove a lever also existed within these two complex protein samples evidently demonstrating the validity of TMP model and leverage theory in MRBE chip.