SU879469A1 - Method of biological article mixture separation - Google Patents

Description

one

The invention relates to physicochemical technology, in particular to methods for the isolation and purification of biological preparations.

There is a known method for the separation and purification of biopreparations by electrophoresis in a free stream 3 in which the separable mixture is injected into the flow of the carrier buffer solution and is subjected to an electric field throughout the length of the flat electrophoretic chamber during the entire separation time. Under the action of the floor, the initial mixture is divided into a series of fractions, which are taken from the stream by a collector.

However, with the known method it is impossible to use the electric field strength above 120 V / cm in the chambers with a larger gap, at which there is a better separation and high-performance purification of biological preparations.

The use of large electric field loads leads to an increase in temperature and the occurrence of significant convective currents, which reduces the efficiency of the method.

The aim of the invention is to improve the accuracy of the method and preserve the viability of the cells.

The goal is achieved next.

The mixture of biological particles is separated by introducing it into the buffer stream and subjecting it to an electric field, while the buffer stream is subjected to an electric field periodically.

An example of the proposed method.

Claims (1)

Electrophoretic separation of the primary suspension of rabbit kidney cells is carried out. The composition of the components of the buffer solution (per 1000 ml):. 1 mmol of triethocolMine P mmol CHjCOOH 2.5 μmol 5 μmol CaC I 5 μM glucose 283 μmol sucrose. 5 mg of albumin of bovine serum 111 the duration of the period of life is chosen from the condition that the temperature gradient does not yet occur in the buffer stream, convective mixing, i.e. by 0.3 mm. Upon reaching such a temperature gradient, the voltage is removed from the electrodes as well. cooling of the buffer solution is continued to its original temperature. The transverse dimensions of the electrophoretic chamber 0-cg are 3,100 mm, where GI is the size of the gap of the chamber slit, mm is the width of the cooled surface of the chamber, mm. The temperature of the chamber is cooled by cooling both surfaces of the chamber. The initial temperature of the buffer solution is 4 ° C, the flow rate of the buffer solution is 1.2 ml / s (the maximum speed of the buffer flow in the center of chamber 4). The voltage on the electrodes of the chamber is 1800 V (the field strength in the chamber is respectively 180 10 V / m. The pulse duration is 2.5 s, the duration of the pause is 10 s. The maximum temperature difference between the 94 most heated part of the buffer flow and the cooled surface of the chamber is The result of separation is 10 fractions, and by the known method b fractions. The preemptive method has such advantages as: it increases the accuracy and efficiency of the free-flow electrophoresis method, expands the possibilities of the free-electrophoresis method single stream, increasing the number of biologics that can be separated; allows the use of high electric field strengths for electrophoretic chambers with a gap greater than 1 mm. Formula of the invention A method for separating a mixture of biological particles by introducing it into a buffer stream field, so that, in order to improve the accuracy of the method and preserve the viability of the cells, the buffer flow is effected by an electric field periodically. Sources of information taken into account in the examination}. K.RannIg, H.WIrth Tree-flow electrophoresis. Theoretfcal and experimental InvestIgation Z. PhysioT. Chem., 1209, 1975.