Electrophoresis method with varying electrical field
A technology of electrophoresis and electric field, applied in the direction of material analysis, measurement device, and instrument by electromagnetic means, which can solve the problems of reducing non-specific binding and unable to completely eliminate it.
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Embodiment 1
[0105] Example 1: Using the electrophoresis method of the present invention to detect the transcription factor Stat5.
[0106] This paper uses the CycloChip developed by the applicant and called CycloChip TM device to illustrate the invention. CycloChips TM is a disposable plastic chip comprising a separation channel and a plurality of electrodes as described herein. An exemplary immunoassay was performed and the term CycloELISA was used to indicate that the chip was used for a purpose other than that for which it was originally designed. Thus, immunoassays (such as ELISAs) that use antibodies to detect analytes effectively use antibodies in a completely different manner.
[0107] Using CycloChips TM Then (as described above), once the analyte is introduced into the channel, a non-uniform electric field in spacetime is applied in the channel. The electric field is essentially a moving wavefront, which moves with velocity k through the separation channel. Like electrophor...
Embodiment 2
[0123] Example 2: Illustrative visualization of analytes using the method of the invention.
[0124] image 3 (Case 1) illustrates the kind of fluorescent signal that can be expected when using the method of the present invention to concentrate the signal over time on the background. In this case, a narrow electric field window between Emax and Emin is used to focus on a specific analyte. moving from left to right, image 3 It was shown that the signal from the antibody / antigen complex (analyte 2) provided a peak at a specific point, whereas the antibody alone (analyte 1) was not concentrated in the window. The graph below is the sum of the signals. Thus, when bound to fluorescently labeled antibodies, a strong signal was obtained over the background. like Image 6 As shown, this can also be used to provide more qualitative and quantitative information by measuring the transient change in fluorescence amplitude over time.
[0125] Figure 4 (Case 2) illustrates the type...
Embodiment 3
[0128] Example 3: Selective collection enrichment
[0129] In conventional electrophoresis, if the user is only interested in part of the sample, the entire separation still needs to be performed. For 2D electrophoresis, this means a week's run. On the contrary, with the present invention, it is possible to set the electric field characteristic and the field parameter k to separate values that achieve a preset transition window. For example, this allows direct isolation of specific biomarkers without isolating any molecules outside the migration window. This is effectively a focusing action, that is, selective collective concentration. The ability to run targeted migration windows allows application specific products.
[0130] Figure 7 A description of selective pool enrichment in a binding assay is provided. Figure 7 The left part of , shows a typical electric field configuration (simple linear field), where Emax = 400V / cm and Emin = 0V / cm. The electric field propag...
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