JPS5952743A - Method and apparatus for imparting hydrogen ion concentration gradient, migrating and dispensing hydrogen ion in noncarrier thin laminar flow isoelectric point electrophresis - Google Patents

Abstract

PURPOSE:To separate minutely a sample liquid into isoelectric point graduation, by arranging a liquid sending side slender tube row for electrophoresis separation along an upper verge of an electrophoresis separation vessel and also, arranging a sample dispensing side slender tube along a lower verge of it, and dispensing by sending the sample liquid and migrating at a sample's own isoelectric point. CONSTITUTION:Whole tube rows 7, 8 consisting of plural slender tubes are arranged facing each other along both upper and lower facing verges of an electrophoresis separation vessel and a liquid for migrating separation is circulated with a pump Po by using the slender tube row 7 for a liquid sending side for migrating separation and also, using the slender tube row 8 for a dispensing side and then, the sample dispensing liquid is dispensed together with the liquid for separation into test tubes 61, 62... by a valve 10 of the dispensing side slender tube row. A synthetic amphoteric electrolyte (an ampholite solution) is used for the liquid for migrating separation and circulated, while conducting electricity by electrodes 5, 5'. After a gradient of hydrogen ion concentration is imparted sufficiently, a sample liquid 4 is sent by a pump P and is dispensed at any time by circulating it.

Description

[Detailed Description of the Invention] A synthetic amphoteric snow melt solution that creates a hydrogen ion concentration gradient by applying electricity is used as the solution, and the sample is sent into an electrophoresis separation tank that creates a hydrogen ion concentration gradient to determine the specific characteristics of the sample. This invention relates to a method and apparatus for electrophoresis and fractionation.

A certain amount of the electrophoretic separation liquid 2 is caused to flow in a laminar flow through an electrophoretic separation tank consisting of two flat plates 1 and 1' placed in parallel. The method of separating and recovering a sample injected from a sample by electrophoresis is known as carrier-free continuous electrophoresis (because no carrier is used in the electrophoresis separation tank), and the principle of this method is shown in Figure 1. An example of the conventional configuration of the device is shown in Fig. 2.
5' is the electrode, Pl, P2... is the preparative pump, P
is a sample injection pump, and 61, 62, . . . are preparative test tubes.

In all of these conventional devices, as shown in Fig. 2, the electrophoretic separation liquid 2 flows in from the inlet 3 provided at the top of the electrophoretic separation tank. Only solutions with a uniform composition can be used as the solution, and it has been impossible to perform isoelectric focusing in which a hydrogen ion concentration gradient (p11 gradient) flows in with these devices.

In the present invention, thin tube rows 7 and 8, which are made up of a plurality of thin tubes, are arranged facing each other along the upper and lower opposing picture planes of an electrophoretic separation tank, and the upper thin tube row 7 is placed on the liquid feeding side for electrophoretic separation. Then, with the thin tube row 8 on the lower edge as the separation side, the electrophoretic separation liquid is also circulated on the side using the pump P [at 7 hours, the test tubes 61, 62...・It is a mechanism that can separate the sample separation liquid that stays in each of the thin tube rows together with the separation liquid.

Hydrogen ion concentration gradient (
A synthetic ampholyte solution (amphorite solution) with a pH gradient) was used. The aerophoresis separation tank is circulated while being energized by the electrode 5'5, and after a sufficient hydrogen ion concentration gradient has been created, the sample solution 4 is pumped by the pump P and circulated through the tank seven or more times, and is then inserted into each capillary at the appropriate time. The sample aliquot liquid remaining in the medium is divided together with the separation liquid into a portion J4'y. Valve 1 provided in each capillary of the side capillary row
0 and fractionated, or use a hydrogen ion concentration gradient (pl(
From the beginning of circulating the synthetic amphoteric electrolyte solution (amphorite solution) for gradient), the sample solution is sent, circulated through the electrophoresis separation tank, and fractionated in the same way.

According to the present invention, even a sample liquid for which one electrophoretic separation is insufficient can be precisely separated into equal points DI specific to the sample components while being circulated, and moreover, it can be easily fractionated in a short time. Also, due to this circulation structure, the electrophoresis tank can be made infinitely small, and after the separation is completed, the liquid in the circulation closed circuit is collected in the tube by reverse flow at 7°C, and the finely fractionated solution is stored at 7°C. It has the advantage of being able to separate all of them in large quantities.

[Brief explanation of the drawing]

Figures 1 and 2 are principle diagrams of an electrophoresis separation tank, Figure 2 is a diagram showing a conventional electrophoresis tank, and Figure 3 is a diagram showing the electrophoresis separation tank according to the present invention. FIG. 3 is a diagram showing a case where electrophoresis is performed in real time. In all figures, components having the same function are indicated by the same numerical symbols. 2 ... Electrophoretic separation liquid 4 ... Sample liquid 5, 5' ... Electrode 7 Electrophoretic separation liquid feeding side thin tube row 8 ... Sample separation side thin tube row P ... Sample liquid feeding pump P...Circulation pump 0 Fig. 1 2 Fig. 2

Claims (2)

[Claims] (1) Improving the pair of electrophoresis separation tanks, orienting the tubule rows consisting of a plurality of tubules along each lower edge so that they face each other, and moving from one tubule row to the sacrotubule row? A synthetic amphoteric electrolyte (amphorite), which is a liquid for electrophoretic separation to which a hydrogen ion gradient (P11 gradient) has been applied by applying electricity, is heated inside the electrophoretic separation tank while applying electricity to the electrophoresis separation tank. is circulated to create a hydrogen ion concentration gradient (
P11 gradient), the sample is pumped, circulated, and migrated to the isoelectric point unique to the sample. This is a method of inorganic continuous electrophoresis, applying a hydrogen ion concentration gradient (pH gradient) during electrophoresis, electrophoresis, and preparative separation. (2) Along the upper edge of both the upper and lower opposing edges of the electrophoresis separation tank, the electrophoresis separation liquid feeding side capillary tube row and the sample liquid feeding capillary tube, and the sample separation side capillary tube row along the lower edge, respectively. Pumps 70 are arranged facing each other and feed liquid from the liquid sending side capillary tube array to the preparative side capillary tube array via the electrophoresis separation tank, and further circulate the liquid to the liquid sending side capillary tube array via the preparative side capillary tube array. 1. An electrophoresis apparatus characterized in that a valve is provided in each thin tube row on the separation side, and a sample liquid remaining in each thin tube is separated together with a separation liquid.