Electrophoresis cell, free-flow electrophoresis device containing it and its uses

Abstract

The invention relates to a free-flow electrophoresis cell for purifying and / or separating an initial solution containing a product to be purified and / or separated, comprising at least one plate X between two adjacent plates Y: X being an electrophoresis plate made of an inert material, Y being a sealed plate made of sapphire or 99% α-Al2O3 alumina, an electrophoresis chamber comprising s subunits Ui, s representing an integer from 2 to 5 and i being an integer from 1 to s, and a cooling circuit, characterized in that the electrophoresis chamber comprises: intermediate channels connecting the two successive subunits. (No figure)

Description

Title of the invention: Electrophoresis cell, free-flow electrophoresis device containing it and its uses

[0001] The present invention relates to a new electrophoresis cell, a new free-flow electrophoresis device with a new configuration and its uses.

[0002] High-performance liquid chromatography (HPLC) and zone electrophoresis using a support are analytical and / or preparative techniques for separating molecules, particularly biomolecules, present in a mixture. However, the need for a stationary phase in HPLC used in industrial processes for separation and / or purification leads to significant costs due to the recycling of various solvents and the regular replacement of solid phases. Furthermore, zone electrophoresis has limitations for industrial-scale use as a preparative method and is primarily used in analytical techniques.

[0003] There is a need to develop a new electrophoresis cell allowing effective and optimized purification and / or separation of products contained in an initial solution by free-flow electrophoresis.

[0004] There is therefore a need to develop an adaptable, robust device without the need for demanding maintenance, operating in continuous flow, designed for easy use and modular implementation, in order to allow purification and / or separation of the expected products in volumes and quantities on an industrial scale.

[0005] One of the aims of the invention is to provide an electrophoresis cell.

[0006] Another object of the invention is to provide a device for the purification and / or separation of molecules by free-flow electrophoresis that can operate in continuous flow and on an industrial scale.

[0007] Another object of the invention is a process for purifying and / or separating molecules, in particular biomolecules, adaptable to an industrial scale.

[0008] A first object of the present invention relates to a free-flow electrophoresis cell for purifying and / or separating an initial solution containing a product to be purified and / or separated, comprising - at least one plate X between two adjacent plates Y:

[0009] X being an electrophoresis plate (1) made of inert material,

[0010] Y being a sealed plate (2) made of sapphire or alumina A12O3 with 99% a-Al2O3, - an electrophoresis chamber comprising s subunits U;, s representing an integer from 2 to 5 and i being an integer ranging from 1 to s, - in which each subunit U; comprises • a hollowed-out section in a closed plate X sandwiched between two plates Y, • in the shape of a rectangular parallelepiped with 4 lateral faces (ai, bi5 Ci, di) and 2 upper and lower faces (e, f), • said hollowed-out part being of length Lo; and of width La;, • of height h; corresponding to the thickness of the plate X-ray electrophoresis, from 25 µm to 20 mm, • the lateral faces (a, b) being parallel to each other, the face (a) being delimited by two edges (Ah, A2) of dimension La, and the face (b) being delimited by two edges (Bh, B2) of dimension La • the lateral faces (c;, d;) being parallel to each other, the face (Ci) being delimited by two edges (Ch, C2;) of dimension Lûi and the face (d;) being delimited by two edges (Dh, D2;) of dimension Lo;, • n; successive entries E;(1), E;(2) to Ei(n;-1), Ei(n;), n; being an integer from 4 to 9, distributed on the face (a,) between Ah and A2; and aligned along a direction parallel to Ah and A2b • m; successive outputs from Si(1), Si(2) to Si(mi-1), Si(mi), m; being an integer from 4 to 12, distributed on the face (b;) between B h and B2; and aligned along a direction parallel to B h and B2;, so that Si(l) faces E;(l) and Si(m;) faces Ei(n;) along a direction parallel to Ch and Dh, - a cooling circuit,

[0011] characterized in that the electrophoresis chamber comprises: - (s-1) intermediate channels Vk, k being an integer ranging from 1 to (s-1),

[0012] each Vk channel linking the two successive subunits Uk and Uk+i

[0013] by one of the outputs SVk(Uk) chosen from Sk(2) to Sk(mk-1) of the subunit Uk to one of the inputs EVk(Uk+i) of the subunit Uk+choisie from Ek+i(2) to Ek+i(nk+rl).

[0014] The Inventors have demonstrated a new electrophoresis cell configuration that optimizes the purification and / or separation of an initial solution containing a product to be purified and / or separated, which may consist of a mixture of compounds of interest. In this cell, the separation and / or purification are effectively implemented, in the sense that the cell allows to recover in its different outlets a solution of the purified and / or separated product or solutions of the compounds of said product which have been purified and / or separated.

[0015] The configuration of the cell of the invention, comprising an electrophoresis chamber made up of subunits connected by intermediate channels, allows for a succession of separations and / or purifications by free-flow electrophoresis by selecting the portion of the flow as the solution to be purified and / or separated, with the possibility of modulating and renewing the free-flow electrophoresis separation conditions independently in each subunit. Furthermore, the cell is adapted to recover, at the outlet of one of the subunits, one of the compounds of interest from the purified and / or separated product and to continue the separation and / or purification of the other compounds of said product in the other subunits.

[0016] This new electrophoresis cell can be integrated into an industrial-sized electrophoresis cell device and used in a continuous flow free-flow electrophoresis purification and / or separation process.

[0017] The electrophoresis cell according to the invention is characterized by the presence of an intermediate channel or intermediate channels linking two subunits of the electrophoresis chamber.

[0018] An electrophoresis chamber of s subunits has (s-1) intermediate channels Vk, k being an integer ranging from 1 to (s-1), each intermediate channel connecting the two successive units Uk and Uk+i.

[0019] Two subunits Uk and Uk+[ are said to be successive when they are connected to each other by at least one intermediate channel Vk, and when, in the circulation of fluids in the electrophoresis chamber, a part of the flux passes from the subunit Uk to the subunit Uk+[ through the channel Vk.

[0020] Each intermediate channel extends from an output of a first subunit, distinct from the two outputs located respectively at the two ends of the alignment of outputs, to an input of the next subunit, distinct from the two inputs located respectively at the two ends of the alignment of inputs.

[0021] The Vk channel extends from an output of the unit Uk, hereafter named SVk(Uk), which is different from the two outputs Sk(l) and Sk(mk) located at the ends of the alignment of outputs, to one of the inputs of the unit Uk+i, hereafter named SEk(Uk+i), which is different from the two inputs Ek+i(l) to Ek+i(nk+i) located respectively at the ends of the alignment of inputs.

[0022] The intermediate channel Vk functions to direct a portion of the flow circulating in the subunit Uk, this flow containing the product to be separated and / or purified or one of the compounds to be purified and / or separated from said product or a mixture of compounds said product or a mixture of said compounds product to be purified and / or separated, towards the Uk+i subunit, this in the absence of an electric field.

[0023] The intermediate channel allows a portion of the outgoing flow circulating from a first subunit to be selected, excluding the two flows corresponding respectively to the liquid cathode and the liquid anode, and the aforementioned portion of this flow to be introduced as a new solution to be purified and / or separated into a subsequent second subunit to allow another separation and / or purification step by free-flow electrophoresis under adaptable conditions which may be identical or different from those of the first subunit.

[0024] These adaptable conditions include the dimensions of the subunit, the electric field, the fluidic flow rate, and the buffer solutions.

[0025] Thus for s equal to 2, the electrophoresis chamber comprises two subunits Ui and U2 and an intermediate channel Vi linking the two subunits Ui and U2.

[0026] For s equal to 3, the electrophoresis chamber comprises three subunits Ui, U2 and U3 and two intermediate channels Vi and V2, Vi linking Ui and U2, V2 linking U2 and U3.

[0027] For s equal to 4, the electrophoresis chamber comprises four subunits Ub U2, U3 and U4 and three intermediate channels Vb V2 and V3, Vi linking Ui and U2, V2 linking U2 and U3, V3 linking U3 and U4.

[0028] For s equal to 5, the electrophoresis chamber comprises five subunits Ui, U2, U3, U4 and U5 and four intermediate channels Vi, V2, V3 and V4, Vi linking Ui and U2, V2 linking U2 and U3, V3 linking U3 and U4, V4 linking U4 and U5.

[0029] The intermediate channel Vi extends from an output SVi(Ui) of the subunit Ui, chosen from the (mr2) outputs of Si(2) to Si(mrl), to one of the inputs EVi(U2) of the subunit U2 chosen from the (n2-2) inputs of E2(2) to E2(n2-1), mi being the number of outputs of Ui and n2 being the number of inputs of U2.

[0030] The intermediate channel V2 extends from an output SV2(U2) of the subunit U2, chosen from the (m2-2) outputs of S2(2) to S2(m2-1), to one of the inputs EV2(U3) of the subunit U3 chosen from the (n3-2) inputs of E3(2) to E3(n3-1), m2 being the number of outputs of U2 and n3 being the number of inputs of U3.

[0031] The intermediate channel V3 extends from an output SV3(U3) of the subunit U3, chosen from the (m3-2) outputs of S3(2) to S3(m3-1), to one of the inputs EV3(U4) of the subunit U4 chosen from the (n4-2) inputs of E4(2) to E4(n4-1), m3 being the number of outputs of U3 and n4 being the number of inputs of U4.

[0032] The intermediate channel V4 extends from an output SV4(U4) of the subunit U4, chosen from the (rrq-2) outputs of S4(2) to S / rru-1), to one of the inputs EV4(U5) of the subunit U5 chosen from the (n5-2) inputs of E5(2) to E5(n5-1), m4 being the number of outputs of U4 and n5 being the number of inputs of U5.

[0033] The term "free-flow electrophoresis" means electrophoresis that does not employ a stationary phase, i.e., without the use of a solid phase to support the migration of species during electrophoresis.

[0034] The "electrophoresis cell" is a component unit of a device. The electrophoresis cell comprises an electrophoresis chamber, including the walls of plates X and Y that constitute and close the electrophoresis chamber, and a fluidic circuit connecting the inlets and outlets of said chamber. This internal fluidic circuit includes supply, recovery, and discharge channels connecting the inlets and outlets of said electrophoresis chamber to circuits outside the cell.

[0035] The term "plate" refers to a rigid element, generally in the form of a rectangular parallelepiped, in which at least two faces are parallel to each other and these faces represent primarily the total area of ​​this element, namely the area of ​​said two parallel faces represents more than half of this total area. These two faces are called "surfaces" or "lower or upper surfaces" of the plate.

[0036] The distance between these two surfaces of the plate defines the thickness of the plate.

[0037] The term "electrophoresis plate" refers to the plate, designated X, containing at least one subunit or a hollowed-out portion of the electrophoresis chamber. Plate X is made of a material inert to electrophoresis.

[0038] By "sealing plate", named Y, we mean a plate intended to ensure the fluidic sealing of the electrophoresis cell.

[0039] The cell comprises at least one plate X located between two plates Y, which ensure the sealing of the electrophoresis chamber of plate X.

[0040] The Y plate is made of inert material, electrically insulating and thermally conductive in order to ensure the chemical and electrical inertness of the device necessary for the free-flow electrophoresis process and in order to ensure thermal conductivity between the cooling circuit and the subunits of the electrophoresis chamber to allow control of the temperature of the electrophoresis cell.

[0041] Plate Y is made of sapphire or alumina A12O3 with 99% a-Al2O3.

[0042] In the preferred embodiment of the invention, the Y plates are made of sapphire or alumina A12O3 with 99% a-Al2O3 preferably in sapphire.

[0043] By "sapphire" is meant a material consisting of corundum or an alumina A12O3 comprising 99% by weight of the α-Al2O3 phase.

[0044] Advantageously, the Y plate has a Mohs hardness of 9 (Coridon), a thermal conductivity of 30 W / m / K to 50 W / m / K. The Y plate has high mechanical resistance, especially compared to a glass plate.

[0045] By way of non-limiting example, the sapphire Y plate is supplied by Saint-Gobain (Luxium Solutions).

[0046] The mechanical resistance of the sapphire plates makes it possible to ensure the sealing of the electrophoresis cell by mechanical tightening of the plates, advantageously, without resorting to the use of a chemical glue to ensure the sealing.

[0047] The sapphire plate also has the advantage of being transparent.

[0048] The introduction of a cooling circuit using sapphire or alumina (Al2O3) plates with 99% a-Al2O3 as a means of separation between the cooling circuit and the subunits of the electrophoresis chamber induces excellent temperature control and homogeneity such that it allows: - a thickness h; subunits constituting the electrophoresis cell can vary from micrometers to millimeters, thus increasing the processing capacity of the solutions to be purified or separated, - purification and / or separation of thermosensitive molecules, - introduction of a temperature gradient in the subunits.

[0049] The nature of the materials of the Y plates, which are sapphire or alumina A12O3 with 99% a-Al2O3, is an important characteristic of the electrophoresis cell of the invention for regulating and controlling the temperature, which is a critical parameter in the free-flow electrophoresis process, in particular for maintaining a laminar flow system, in addition to the aspect of protein denaturation.

[0050] The electrophoresis cell comprises at least one sequence of YXY plates, plate X being located between two adjacent Y plates. By "adjacent plates" is meant plates directly joined by their surface.

[0051] The electrophoresis cell comprises a single YXY sequence when all subunits are located on the same plate. It can comprise up to 5 different YXY plate sequences when all subunits are distributed on different X plates, allowing the subunit height h to be adjusted according to the operating conditions for separation and / or purification. The number of YXY sequences is less than s when at least two subunits are located in the same plate.

[0052] The "electrophoresis chamber" is the part of the electrophoresis cell in which the circulation and separation of the product to be purified and / or separated take place.

[0053] In the present invention, the electrophoresis chamber is made up of multiple subunits connected to each other by intermediate channels.

[0054] By "a subunit of the electrophoresis chamber" is meant a part of an electrophoresis plate X comprising:

[0055] - a hollowed-out portion in the X-ray electrophoresis plate, delimited by walls lateral and wall surfaces of plates Y adjacent to the aforementioned plate X closing the hollowed-out part, and

[0056] - the inlets and outlets located in the side walls of said hollowed-out part.

[0057] The subunits of the electrophoresis chamber form the space in which free-flow electrophoresis takes place.

[0058] The number of subunits "5" is at least two, it can vary from 2 to 5, namely from 2, 3, 4 and 5, but it can also be beyond 5, between 6 and 10, for example from 6, 7, 8, 9 or 10 when the separation conditions, in particular flow rate and yield of the product to be purified and / or separated, are appropriate.

[0059] Each subunit U; in which free-flow electrophoresis takes place, comprises a hollowed-out portion of a plate X, closed by the lateral walls in the plate X and by the upper and lower walls of the surfaces of the two adjacent plates Y, said hollowed-out portion being in the general shape of a rectangular parallelepiped.

[0060] The term "in the shape of a rectangular parallelepiped" means that this hollowed part is generally inscribed within a rectangular parallelepiped of length Lo;, width La; and height h; corresponding to the thickness of the plate X. In defining the general shape of the hollowed part, no account is taken of any means possibly present in the electrophoresis chamber, such as the channeling means present at the inlets and outlets and the protrusions.

[0061] The rectangular parallelepiped comprises four lateral faces (a;, b;, Ci, d;) and two faces (e;, h), respectively lower and upper.

[0062] The lateral faces (ai5 b;) are parallel to each other and of the same dimensions.

[0063] The face (a;) is delimited by two edges (Ah, A2;) of dimension La;, spaced by h;.

[0064] The face (b;) is delimited by two edges (B h, B2;) of dimension La;, spaced by h;.

[0065] The lateral faces (c;, d;) are parallel to each other and of the same dimensions.

[0066] The face (cO is delimited by two edges (Ch, C2;) of dimension Lo;, spaced by h;.

[0067] The face (d;) is delimited by two edges (Dh, D2;) of dimension Lo;, spaced by h;.

[0068] The upper (e;) and lower (h) faces are parallel to each other and of the same dimensions. They constitute the surfaces framing the hollowed-out part, cut into the plate X.

[0069] The face (e;) is delimited by the edges (Ah, B h, Ch, Dh) forming a rectangle of width La; and of length Lo;.

[0070] The face (h) is delimited by the edges (A2;, B2;, C2;, D2;) forming a rectangle of width La; and of length Lo;.

[0071] The height h; corresponds to the thickness of the plate X, and thus to the height of the subunit U; and of its hollowed part.

[0072] Two subunits of the chamber can be of different heights when their hollowed part are respectively in two different YXY plate sequences.

[0073] The height h; is from 25 pm to 20 mm.

[0074] The range from 25 µm to 20 mm includes the following ranges: from 25 to 50 µm; from 50 to 75 µm; from 75 to 100 µm; from 100 to 200 µm; from 200 to 300 µm; from 300 to 400 µm; from 400 to 500 µm; from 500 to 600 µm; from 600 to 700 µm; from 700 to 800 µm; from 800 to 900 µm; from 900 µm to 1.0 mm; from 1.0 to 2.0 mm; from 2.0 to 3.0 mm; from 3.0 to 4.0 mm; from 4.0 to 5.0 mm; from 5.0 to 6.0 mm; from 6.0 to 7.0 mm; from 7.0 to 8.0 mm; from 8.0 to 9.0 mm; from 9.0 to 10.0 mm; from 10.0 to 11.0 mm; from 11.0 to 12.0 mm; from 12.0 to 13.0 mm; from 13.0 to 14.0 mm; from 14.0 to 15.0 mm; from 15.0 to 16.0 mm; from 16.0 to 17.0 mm; from 17.0 to 18.0 mm; from 18.0 to 19.0 mm; from 19.0 to 20.0 mm.

[0075] Advantageously the height h; is from 25 to 200 pm.

[0076] Advantageously the height h; is from 1.0 to 5.0 mm.

[0077] The dimensions of the subunits are independent of each other, namely the width La;, the length Lo; and the height h; of each subunit U; can be identical or different from each other.

[0078] Similarly, the number of inputs n; and the number of outputs m; of the subunits Ui can be identical or different from each other.

[0079] Thus, for the "5" subunits, these dimension parameters and numbers of inputs and outputs (La;, Lo;, h;, n;, m; ) of each subunit U;, i being an integer ranging from 1 to s, can be adapted and configured in the cell to optimize the purification or separation of a product in an initial solution.

[0080] By "inlet" is meant a passage allowing the flow of a liquid fluid from the outside to the inside of a system containing a closed hollow part, configured to contain said fluid, for example such as the U subunits of the electrophoresis chamber.

[0081] By "outlet" is meant a passage allowing the flow of a liquid fluid from the inside to the outside of a system containing a closed hollow part, configured to contain said fluid, for example such as the U subunits; of the electrophoresis chamber.

[0082] On face (a;), the n; inputs into the hollowed-out part of a subunit U; of the electrophoresis chamber are distributed successively, respectively referenced E;(l) to E; (n), i.e. Ej(l), Ej(2) to E(nrl), E^n;). The number of inputs n; varies from 4 to 9, i.e. 4, 5, 6, 7, 8 and 9, preferably 5 or 6.

[0083] The entries E;(l) to Ei(n;) are located between the edges Al; and A2; and aligned along a direction parallel to Al; and A2;, advantageously at a substantially equal distance from each other.

[0084] By "aligned" is meant that the inlets, respectively the outlets, are spatially close to the same straight line. The inlets can therefore be positioned slightly in front of or behind the face (a;) and / or slightly above or below each other. The positioning of the inlets is configured to introduce the different fluxes into the subunit U; of the electrophoresis chamber in such a way as to allow their circulation in said subunit U; parallel to the edge Cl;.

[0085] On face (b;), the m; outputs are distributed successively, respectively referenced S;(l) to S^mO, i.e. S;(l), S;(2) to Si(m;-1), S^mO. The number of outputs m; varies from 4 to 12, i.e. 4, 5, 6, 7, 8, 9, 10, 11 and 12, preferably 5 or 7.

[0086] The entries S;(l) to Si(m;) are located between the edges Bl; and B2; and aligned along a direction parallel to B h and B2;, advantageously at a substantially equal distance from each other.

[0087] The entries E;(l) and E; (n;) are located near the two ends of the edges Al; and A2;, that is to say as close as possible to the faces (c;) or (d;).

[0088] The outputs S;(l) and Si (m,) are located near the two ends of the edges Bl; and B2;, that is to say as close as possible to the faces (cO or (d;).

[0089] Thus, for each subunit Ui, the inlet E;(l) is configured for the introduction of a liquid cathode CAL(i). The outlet S;(l) is configured for the evacuation of said liquid cathode after circulation in the subunit U; of the electrophoresis chamber.

[0090] The inlet E;(n) is configured for the introduction of a liquid anode ANL(i). The outlet Si(m;) is configured for the evacuation of said liquid anode after circulation in the subunit U; of the electrophoresis chamber.

[0091] The inlets and outlets of a subunit U; of the electrophoresis chamber are configured so that E;(l) faces Si(l) and Ei(n;) faces Si(m;) so as to induce the electric field CE(i) in the subunit U; of the electrophoresis chamber between the liquid anode ANL(i) and the liquid cathode CAL(i) during the circulation of the fluids in order to allow the implementation of an electrophoresis step.

[0092] The liquid anode and the liquid cathode are electrolyte solutions, that is to say solutions comprising ions.

[0093] By "liquid cathode" is meant an electrolyte solution configured to act as a cathode during electrophoresis.

[0094] By "liquid anode" is meant an electrolyte solution configured to act as an anode during electrophoresis

[0095] During operation in fluid circulation, the presence of the liquid cathode CAL(i) and the liquid anode ANL(i) generates an electric field in the subunit U; of the electrophoresis chamber when the electrolytic solutions are charged.

[0096] The liquid cathode and the liquid anode are electrolytic solutions, capable of generating an electric field, which can be increased or decreased by action of the generator which charges the liquid anode and the liquid cathode and / or with an increase in the concentration of ions.

[0097] Thus the electric fields generated CE(i) in the subunits Upeuvent be identical or different from one subunit to another depending on the liquid anode and the liquid cathode in circulation.

[0098] Function of the inputs and outputs of the subunit U i

[0099] In the first subunit Ui, one of the inlets Ei(2) to Ei(nrl) is configured for the introduction of the initial solution to be purified and / or separated into the first subunit Ui of the electrophoresis chamber.

[0100] At least one of the inputs Ei(2) to Ei(nrl), distinct from the aforementioned input of the initial solution to be purified and / or separated, is configured for the introduction of at least one buffer solution into the subunit Ui in order to allow free-flow electrophoresis from the initial solution in the presence of at least one buffer solution under an electric field CE(1) generated by the circulation of the liquid cathode and anode.

[0101] This subunit Ui of ni entries therefore contains (nr3) remaining entries dedicated to the introduction of one or more buffer solutions of identical or different compositions, in addition to the entries dedicated to the liquid electrodes Ei(l) and E|(nJ and that dedicated to the introduction of the initial solution to be purified and / or separated.

[0102] The outputs Si(1) and Si(mi) are dedicated to the respective circulation of the two electrodes liquids.

[0103] At least one of the outputs Si(2) to Ei(mrl), named SVi(Ui) is dedicated to the intermediate channel Vi linking the subunit Ui to the subunit U2.

[0104] At least one of the remaining outlets is dedicated either to recovering a stream of one of the compounds present in the product to be purified and / or separated, or to evacuating a buffer solution that has circulated in the Up subunit

[0105] Function of the inputs and outputs of the subunit U t

[0106] In each subunit U, different from Ui and Us, at least one of the inlets E(2) to Edn-1 is connected at least to the intermediate channel V, which allows a portion of the flux from the subunit Um, containing the purified and / or separated product or a mixture of the compounds constituting said product, to be recovered and introduced into the subunit Ui, without the presence of an electric field, as a solution to be purified and / or separated in order to perform a polishing step, namely a purification and / or separation step, by electrophoresis in the subunit Up

[0107] At least one other of the inputs E;(2) to E(n -1), distinct from the aforementioned input connected to the Vu channel, is configured for the introduction of at least one buffer solution into the subunit U; in order to permit free-flow electrophoresis from said solution to be purified and / or separated in the presence of at least one buffer solution under an electric field CE(i) generated by the circulation of the cathode CAL(i) and the anode (ANL(i) liquids.

[0108] This subunit U; of n; entries therefore contains (nr3) remaining entries dedicated to the introduction of one or more buffer solutions of identical or different compositions, in addition to the entries dedicated to the liquid electrodes E;(l) and E^nO and that dedicated to the V^ channel.

[0109] The outlets Si(l) and Si(m;) are dedicated to the respective circulation of the two liquid electrodes.

[0110] At least one of the outputs S;(2) to Ei(mi-1), named SVi(Ui) is dedicated to the intermediate channel V; linking the subunit U; to the subunit Ui+i.

[0111] At least one of the remaining outlets is dedicated either to the recovery of a stream of one of the compounds present in the product to be purified and / or separated, or to the evacuation of a buffer solution that has circulated in the U subunit;.

[0112] Function of the inputs and outputs of the subunit U s

[0113] In the Us subunit, at least one of the inlets Es(2) to Es(ns-1) is connected at least to the intermediate channel V, which allows to recover a part of the flux containing the purified and / or separated product or of a mixture of the compounds constituting said product, said part being from the Us subunit which has been purified and / or separated and (s-2) times polished and to introduce it into the Us subunit, without presence of electric field, as a solution to be purified and / or separated in order to be able to carry out a polishing step (purification and / or separation step) by electrophoresis.

[0114] At least one other of the inputs Es(2) to Es(ns-1), distinct from the aforementioned input connected to the Vs.i channel, is configured for the introduction of at least one buffer solution into the Us subunit in order to permit free-flow electrophoresis from said solution to be purified and / or separated in the presence of at least one buffer solution under an electric field CE(s) generated by the circulation of the liquid cathode CAL(s) and the liquid anode (ANL(s).

[0115] This Us subunit of ns inputs therefore contains (ns-3) remaining inputs dedicated to the introduction of one or more buffer solutions of identical or different compositions, in addition to the inputs dedicated to the liquid electrodes Es(l) and Es(ns) and the one dedicated to the intermediate channel

[0116] In the last Us subunit of the electrophoresis cell, at least one of the Ss(2) to Ss(ms-1) outputs is configured for the recovery of the purified solution and / or separated after the electrophoresis process carried out in the electrophoresis chamber.

[0117] The Ss(l) and Ss(ms) outlets are dedicated to the respective circulation of the two liquid electrodes.

[0118] In the electrophoresis cell of the invention, as described above, the inputs and outputs of each subunit have distinct functions:

[0119] - the inlets and outlets at the ends are configured to allow circulation liquid electrodes to generate an electric field across the entire subunit,

[0120] - an inlet is dedicated to the introduction of the solution to be purified and / or separated or said to polish,

[0121] - the other entry(ies) are dedicated to the introduction of one or more solutions buffer,

[0122] - in the subunits Ui to Us.i, at least one of the outputs, distinct from those of the electrolytes, is dedicated to recovering a portion of the stream containing the solution to be purified and / or separated in the next subunit.

[0123] - in all subunits at least one of the outputs, distinct from that of the electrolytes, is dedicated either to the recovery of a compound from the product to be purified and / or separated, or to the disposal of the buffer solution after circulation.

[0124] - in the Us subunit, at least one of the outputs, distinct from that of the electrolytes, is dedicated to recovering the purified and / or separated product or one of the compounds of said product and another outlet is dedicated to evacuating the buffer solution.

[0125] The cell according to the invention may include supply channels for the liquid electrodes (anode and cathode), one or more buffer solutions, a supply channel for the product to be purified and / or separated into Ui cells, and at least one output recovery channel in Us.

[0126] According to a particular embodiment, the present invention relates to a cell as defined above, comprising: - supply channels configured to connect the 5 inputs E;(l), i varying from 1 to s, of each subunit of the electrophoresis chamber to at least one external micro / milli fluidic circuit supplying at least one liquid cathode, - supply channels configured to connect the E;(ni) inputs of each subunit of the electrophoresis chamber to at least one external micro / milli fluidic circuit supplying at least one liquid anode, - a supply channel configured to connect at least one of the inputs Ei(2) to Ei(nrl) of the subunit Ui to an external micro / milli fluidic circuit for supplying an initial solution containing a product to be purified and / or separated, - supply channels configured to connect at least one inlet of each subunit of the electrophoresis chamber to at least one external micro / millifluidic circuit supplying at least one buffer solution, - at least one recovery channel configured to connect one of the outputs of the Us subunit chosen from Ss(2) to Ss(ms-1) of the electrophoresis chamber to an external micro / milli fluidic recovery circuit.

[0127] The inlets and outlets of the electrophoresis chamber are those connected to the outside of the cell, i.e., those not connected to intermediate channels linking the subunits. These inlets and outlets are connected by supply, recovery, or discharge channels to external supply, recovery, or discharge circuits configured for micro- or millifluidic circulation of the fluid flows.

[0128] The term "supply channel" means a channel connected to an inlet, adapted to supply a subunit of the chamber with flow.

[0129] The term "recovery channel" means a channel connected to an output, adapted to recover at least one stream of interest, namely containing the product or one of its compounds.

[0130] The term "evacuation channel" means a channel connected to an outlet, adapted to evacuate a flow that has circulated in the chamber.

[0131] The term “microfluidic circuit” means a set of channels with a cross-sectional dimension on the order of a micrometer.

[0132] The term "millifluidic circuit" means a set of channels with a cross-sectional dimension on the order of a millimeter.

[0133] These supply, recovery or evacuation channels are advantageously partially or totally engraved in the plates X.

[0134] Liquid electrolyte supply and discharge channels

[0135] The electrophoresis cell containing s subunits comprises s inputs E;(l) and s outputs Si( 1 ), i varying from 1 to s.

[0136] Each input E;(l) is linked to a supply channel of a liquid cathode CAL(i). These s supply channels are connected to at least one external supply circuit of at least one solution constituting the liquid cathode. Other external supply circuits of a liquid cathode, up to a number of s circuits, can be linked to these inputs when different CE(i) electric fields are sought.

[0137] Each outlet S;(l) is linked to a liquid cathode evacuation channel CAL(i).

[0138] The electrophoresis cell containing s subunits comprises s inputs Ei(n;) and s output S^m;), i varying from 1 to s.

[0139] Each input Ei(n;) is linked to a supply channel of a liquid anode ANL(i). These s supply channels are connected to at least one external supply circuit of at least one solution constituting the liquid anode. Other external supply circuits for a liquid anode, up to a number s circuits, can be linked to these inputs when different electric fields CE(i) are required.

[0140] Each outlet S^m;) is linked to a liquid anode evacuation channel ANL(i).

[0141] Supply channels for buffer solution(s) for electrophoresis

[0142] In the subunit Ub, at least one of the inputs Ei(2) to Ei(nrl) is connected by a supply channel to an external micro / millifluidic circuit supplying an initial solution containing a product to be purified and / or separated. The remaining (nr3) inputs, with at least one remaining input ni being greater than 4, are connected by supply channels to one or more buffer solution(s).

[0143] In the other subunits U2 to Us, the (n;-3) inputs of the subunit U; which are distinct from the two inputs located respectively at the ends and distinct from the input dedicated to the intermediate channel, are linked respectively by supply channels to one or more buffer solution(s).

[0144] Channels for recovering the product or product compounds or for evacuating buffer solutions

[0145] In the last subunit Us>at least one of the outlets Ss(2) to Ss(ms-1) is linked to a recovery channel connected to an external micro / milli fluidic circuit for the recovery of the purified and / or separated product and at least one of these outlets is linked to a discharge channel for a buffer solution that has circulated in Us.

[0146] In the other subunits Uià Us.i, for (m;-3) outputs of the subunit U; which are distinct from the two outputs located respectively at the ends and distinct from the output dedicated to the intermediate channel connecting the subunits, at least one of these outputs is: - either linked to a recovery channel, advantageously for recovering a purified and / or separated compound constituting said product, - either linked to a discharge channel of a buffer solution that has circulated in U;.

[0147] These supply, recovery and evacuation channels fulfill the distinct function of each inlet and outlet of each subunit described above.

[0148] The cell advantageously contains supply and discharge channels (distinct from those of the liquid electrodes) which are present for at least one buffer solution so as to effectively allow the separation and / or purification by free-flow electrophoresis of the product to be purified and / or separated contained in a solution.

[0149] According to a particular embodiment, the present invention relates to a cell as defined above, said cell being configured to: • in each subunit U;,, in the presence of an electric field generated CE(i) between a liquid cathode CAL(i) and a liquid anode ANL(i) parallel to Ah and perpendicular to CL, • and in operation,

[0150] a) in the Ui subunit: • circulate the liquid cathode CAL(l) from the inlet Ei(l) to the outlet Si(l), • circulate the liquid anode ANL(l) from the inlet Ei(nO) to the outlet Si(mi), • circulate the initial solution containing the product to be separated and / or purified and at least one buffer solution in the Ui subunit, between the liquid cathode CAL(l) and the liquid anode ANL(l), from the inlets Ei(2) to Ei(nr 1) to the outlets Si(2) to Si(mrl), to obtain the purified and / or separated product from the Ui subunit,

[0151] b) in each intermediate channel Vk, k varying from 1 to (s-1), • recover at the SVk(Uk) outlet the purified and / or separated product from the Uk subunit; • to circulate in channel Vk the aforementioned product from the output SVk(Uk) of the subunit Uk, from SVk(Uk) to the input EVk(Uk+i) of the subunit Uk+i,

[0152] c) in the subunit Uk+i: • circulate the liquid cathode CAL(k+l) from the inlet Ek+i(l) to the outlet Sk+[ (1), • circulate the liquid anode ANL(k+l) from the inlet Ek+i(nk+i) to the outlet Sk+i (mk+i), • circulate said purified and / or separated product from the SVk(Uk) outlet of the Uk subunit and at least one buffer solution in the Uk+i subunit, between the liquid cathode CAL(k+l) and the liquid anode ANL(k+l), from the Ek+i(2) to Ek+i(nk+rl) inlets to the Sk+i(2) to Sk+i(mk+rl) outlets,

[0153] d) recover at one of the outputs Ss(2) to Ss(ms-1) the purified and / or separated product.

[0154] The Ui subunit is configured to perform a first step of separation of the product to be separated and / or purified in an initial solution by free-flow electrophoresis under an electric field CE(1) generated by the circulation of the liquid cathode and liquid anode and in the presence of at least one buffer solution.

[0155] The following successive subunits are configured to refine the separation, i.e. "polish", by successive purification and / or separation steps, called "polishing" steps, carried out by free-flow electrophoresis under an electric field CE(i) in the presence of at least one other buffer solution, on a part of the outgoing flow from the previous subunit through an intermediate channel linking the two subunits.

[0156] The subunits are also advantageously configured to be able to recover at the output of one of the subunits U; different from the last subunit Us, an outgoing stream containing one of the compounds of the product to be separated and / or purified and to continue the separation and / or purification in the successive subunits of the other outgoing streams containing the product.

[0157] Complementary Intermediate Channel W

[0158] According to a particular embodiment, the present invention relates to a cell as defined above, further comprising:

[0159] at least one complementary intermediate channel W

[0160] said complementary intermediate channel Wrelie two successive subunits Uq and Uq+i, q being an integer from 1 to (s-1),

[0161] said channel W extending: - from one of the outputs of the unit Uq chosen from Sq(2) to Sq(mq-1), different from the output linked to the channel Vq, - at the input linked to the Vq channel of the Uq+i subunit,

[0162] the aforementioned input of the Vq channel being connected to at most (mq-3) output(s) chosen from Sq (2) to Sq(mq-1).

[0163] In this embodiment, the electrophoresis chamber comprises, between two successive subunits, in addition to the intermediate channel V connecting them, another complementary intermediate channel W connecting the two subunits, extending from one of the outputs of the first subunit, distinct from the three outputs of the cathode, the anode, and the aforementioned intermediate channel V, respectively, to the same input as that of the aforementioned intermediate channel V of the second subunit. The complementary intermediate channel W can be located between any combination of two successive subunits of the electrophoresis chamber, q being any integer from 1 to (s-1).

[0164] This complementary intermediate channel W allows the collected portion of the flow in the first subunit to be expanded, intended to be introduced as a solution to be purified and / or separated in the subsequent second subunit. This is particularly advantageous when the flow containing the desired product or compound spreads between two adjacent outlets of the first subunit.

[0165] Advantageously, in a particular embodiment, the output corresponding to channel V may not be adjacent to that of channel W. This configuration makes it possible to recover in an output of the first subunit located between that of channel V and that of channel W, a stream containing the purified product or a compound separated from this product and to continue in the second following subunit the purification and / or separation with the streams collected by the intermediate channels V and W containing a product not sufficiently purified or a mixture of compounds of the product.

[0166] The subunits are designed to purify and / or separate a product circulating in the flow between the two electrodes in the presence of at least one buffer solution, and the intermediate channels are designed to collect a portion of said flow circulating between the liquid electrodes. Consequently, the inlet connected to the Vq channel of a Uq+i subunit can only be connected to a maximum of (mt| -3) Uq outputs, where mq is the number of outputs of the Uq subunit, because the two outputs dedicated to electrolytes are excluded, as well as one output dedicated either to the recovery of a purified and / or separated product or compound, or to the removal of the buffer solution that has circulated in the Uq unit. Indeed, there would be no benefit in introducing the electrolyte solutions and the entire outgoing Uq flow into the second subunit through this inlet.

[0167] According to a particular embodiment, the present invention relates to a cell as defined above, further comprising:

[0168] R complementary intermediate channel(s), from Wj to WR, R being an integer from 1 to Q:

[0169] [Math.l] Q = ( — :¾ — 4(s ■- 1) and Q > 0

[0170] when r varies from 1 to R,

[0171] the complementary intermediate channel Wr links the subunit Uqr to the successive subunit Uqr+i, qr being an integer from 1 to (s-1),

[0172] said complementary intermediate channel Wr extends: - from one of the outputs of the unit Uqr chosen from Sqr(2) to Sqr(mqr-1) different from the output linked to the channel Vqr, - at the input of EVqr(Uqr+i) linked to the Vqr channel of the subunit Uqr+i,

[0173] the aforementioned input EVqr(Uqr+i) being connected to at most (mqr-3) output(s) chosen from Sqr (2) to Sqr(mq-1).

[0174] In this embodiment, the number Q represents the maximum number of complementary intermediate channels W that can be introduced. It corresponds to the total sum of the number of chamber outlets, subtracting the number of outlets of the last subunit that are not connected to intermediate channels and subtracting, for the (s-1) spacings between two successive subunits, the four outlets dedicated respectively to the two liquid electrodes, the intermediate channel V and the recovery of a product or the evacuation of a buffer solution.

[0175] A chamber can contain R complementary intermediate channels W, with R being an integer from 1 to Q.

[0176] These complementary intermediate channels are named Wr, r varying from 1 to R.

[0177] The complementary intermediate channel Wr is associated with two successive subunits Uqr and Uqr+i, qr being any integer from 1 to (s-1), namely that there is no link between the index r of the channel and the index of the subunits qr.

[0178] This being subject, as previously indicated, to the input linked to the Vqr channel of a subunit Uqr+i can only be linked at most to (mqr -3) outputs of Uqr, mqr being the number of outputs of the subunit Uqr.

[0179] According to a particular embodiment, the present invention relates to a cell as defined above, comprising R complementary intermediate channel(s) Wi to WR, R being an integer from 1 to (s-1), and in which said channels Wi to WR each connect two different successive subunits.

[0180] In this embodiment, the W channels of the electrophoresis chamber each connect two different successive subunits, so the chamber comprises at most one W channel per two successive units.

[0181] Cell of subunits of the same height

[0182] According to a particular embodiment, the present invention relates to a cell as defined above, comprising a plate X between two adjacent plates Y, said plate X comprising the 5 hollowed parts of the subunits U; of said electrophoresis chamber, the height of the hollowed part of each subunit being of identical value.

[0183] In this embodiment, since all the hollowed parts of the subunits are located on the same plate and have the same height h, the electrophoresis cell comprises only one plate X of height h.

[0184] Cell of subunits of different height

[0185] According to a particular embodiment, the present invention relates to a cell as defined above, comprising at least two plates X, each plate X being between two adjacent plates Y,

[0186] each plate X comprising at least one hollowed-out portion of a subunit U; of said electrophoresis chamber.

[0187] This particular embodiment corresponds to a cell configuration in which at least two subunits of the electrophoresis chamber are not on the same plate and may have different heights h. The cell thus comprises at least two different YXY sequences, with plates X that may be of different heights.

[0188] According to a particular embodiment, the present invention relates to a cell as defined above, comprising a plate X between two adjacent plates Y, said plate X comprising the 5 hollowed-out parts of the U subunits; of said electrophoresis chamber, the height of the hollowed-out part of each subunit being of identical value,

[0189] or comprising at least two plates X, each plate X being between two adjacent plates Y, each plate X comprising at least a hollowed-out portion of a subunit U; of said electrophoresis chamber.

[0190] Z. Cooling plates

[0191] According to a particular embodiment, the present invention relates to a cell as defined above, in which said cooling circuit comprises at least one cooling plate Z comprising a heat transfer system, said plate Z being adjacent to one of the two plates Y adjacent to said plate X.

[0192] According to a particular embodiment, the present invention relates to a cell as defined above, in which said a cooling circuit comprises at least two plates Z comprising a heat transfer system, said cell comprising the following sequence of plates ZYXYZ, in particular the sequence YZYXYZY.

[0193] According to a particular embodiment, the present invention relates to a cell as defined above, in which said heat transfer system is a network of pipes configured to allow the circulation of one or more heat transfer fluids parallel to the edges C; of the subunits U;.

[0194] In this embodiment, the heat transfer fluid flows in the same direction as the electrophoresis flow in the chamber, facilitating temperature regulation, which is important for controlling electrophoretic mobility. Advantageously, the piping network can be adapted to generate a temperature gradient.

[0195] According to a particular embodiment, the present invention relates to a cell as defined above, in which said heat transfer system is a network of pipes configured to allow the circulation of one or more heat transfer fluids perpendicular to the edges C; of the subunits U;.

[0196] In this embodiment, the heat transfer fluid is perpendicular to the direction of the electrophoresis flow in the chamber. In this configuration, the inlets and outlets are perpendicular to the inlet and outlet alignments of the subunits, thus simplifying the size of the cell's fluid circuits by distributing the inlets and outlets.

[0197] According to a particular embodiment, the present invention relates to a cell as defined above, in which said heat transfer system is a network of pipes configured to allow the circulation of one or more heat transfer fluids parallel to the edges Ci or perpendicular to the edges Ci of the subunits U;,

[0198] in particular said piping network comprising inlets and outlets of the heat transfer system, engraved in plate Z.

[0199] According to a particular embodiment, the present invention relates to a cell as defined above, comprising at least one cooling plate Z including a heat transfer system, said plate Z being adjacent to one of the two plates Y adjacent to said plate X,

[0200] and / or comprising at least two plates Z comprising a heat transfer system, said cell comprising the following plate sequence ZYXYZ, in particular the sequence YZYXYZY,

[0201] in particular wherein said heat transfer system is a network of pipes configured to allow the circulation of one or more heat transfer fluids parallel to the edges Ci or perpendicular to the edges Cides subunits Ui,

[0202] in particular said piping network comprising inlets and outlets of the heat transfer system, engraved in plate Z.

[0203] Channeling means

[0204] According to a particular embodiment, the present invention relates to a cell as defined above, in which the subunits U; of the electrophoresis chamber comprise means for channeling the flows opening onto the inlets E;(l) to Ei(n;) and / or onto the outlets Si(l) to Si(m;) of the subunits U; of the electrophoresis chamber,

[0205] said channeling means being preferably engraved in the X-ray electrophoresis plate,

[0206] said means being in particular in the form of a triangle, situated between two adjacent inputs or outputs of a subunit.

[0207] These channeling means are configured to allow the orientation of the flows in the subunit of the electrophoresis chamber at the level of each of the inlets and outlets, in order to better distribute the flows in the case of the inlets over the entire width of the hollowed part and to better concentrate the flows in the case of the outlets.

[0208] Advantageously, these channeling means are an integral part of the electrophoresis chamber subunit, i.e., fused with the walls of the electrophoresis chamber subunit and are made of the same material as the electrophoresis plate. These channeling means are located along the faces (a;) and (b;) of the rectangular parallelepiped in which the hollowed-out portion of the subunit U; is inscribed.

[0209] Channeling means are, for example, triangular or beveled point-shaped elements situated between two inlets or two outlets.

[0210] Dimensions of subunits

[0211] According to a particular embodiment, the present invention relates to a cell as defined above, in which

[0212] the width La; of each of the s subunits U; is from 1.0 to 8.0 cm,

[0213] and / or the length Lo; of each of the s subunits U; of the electrophoresis chamber is 2.0 to 20.0 cm.

[0214] The range of "1.0 to 8.0 cm" includes the ranges: from 1.0 to 2.0 cm; from 2.0 to 3.0 cm; from 3.0 to 4.0 cm; from 4.0 to 5.0 cm; from 5.0 to 6.0 cm; from 6.0 to 7.0 cm; from 7.0 to 8.0 cm.

[0215] The range of "2.0 to 20.0 cm" includes the following ranges: from 2.0 to 3.0 cm; from 3.0 to 4.0 cm; from 4.0 to 5.0 cm; from 5.0 to 6.0 cm; from 6.0 to 7.0 cm; from 7.0 to 8.0 cm; from 8.0 to 9.0 cm; from 9.0 to 10.0 cm; from 10.0 to 11.0 cm; from 11.0 to 12.0 cm; from 12.0 to 13.0 cm; from 13.0 to 14.0 cm; from 14.0 to 15.0 cm; from 15.0 to 16.0 cm; from 16.0 to 17.0 cm; from 17.0 to 18.0 cm; from 18.0 to 19.0 cm; from 19.0 to 20.0 cm

[0216] X-Plate Material

[0217] According to a particular embodiment, the present invention relates to a cell as defined above, in which the X-ray electrophoresis plate is made of a material selected from polytetrafluoroethylene (PTFE), perfloroalkoxy (PFA) and fluoroethylene propylene (FEP), in particular Teflon™, Teflon™-PFA and Teflon™-FEP plates.

[0218] According to a particular embodiment, the present invention relates to a cell as defined above, in which the subunits U; of the electrophoresis chamber comprise means for channeling the flows opening onto the inlets E;(l) to Ei(n;) and / or onto the outlets Si(l) to Si(m;) of the subunits U; of the chamber electrophoresis, the said channeling means preferably being engraved in the electrophoresis plate X,

[0219] said means being in particular triangular in shape, situated between two adjacent inputs or two adjacent outputs of a subunit,

[0220] and / or wherein the width La; of each of the s subunits U; is from 1.0 to 8.0 cm, and / or the length Lo; of each of the s subunits U; of the electrophoresis chamber is from 2.0 to 20.0 cm,

[0221] and / or wherein the X electrophoresis plate is made of a material selected from polytetrafluoroethylene (PTFE), perfloroalkoxy (PFA) and fluoroethylene propylene (FEP), in particular Teflon™, Teflon™-PFA and Teflon™-FEP plates.

[0222] Two-subunit electrophoresis cell

[0223] According to a particular embodiment, the present invention relates to a cell as defined above, in which 5 equals 2, the electrophoresis chamber comprising two subunits Ui and U2 - The Ui subunit includes • a hollowed-out section in a plate X closed by two plates Y • in the shape of a rectangular parallelepiped with 4 lateral faces (ab bb Ci, di) and 2 upper and lower faces (eb fi), • said hollowed-out part being of length Loi and width Lai • of height hi corresponding to the thickness of the plate X-ray electrophoresis, from 25 µm to 20 mm, • the lateral faces (ab bj being parallel to each other, the face (ai) being delimited by two edges (Alb A2i) of dimension Lai and the face (bi) being delimited by two edges (Bli, B2i) of dimension Lai, • the lateral faces (ci, di) being parallel to each other, the face (cj) being delimited by two edges (Clb C2i) of dimension Law and the face (dj) being delimited by two edges (Dli, D2i) of dimension Law, • neither successive entries Ei(1), Ei(2) to Ei(nrl), Ei(ni), nor being an integer from 4 to 9, distributed on the face (ai) between Ali and A2i and aligned along a direction parallel to Ali and A2b • mi successive outputs from Si(1), Si(2) to Si(mrl), Si(mi), mi being an integer from 4 to 12, distributed on the face (bi) between Bh and B2i and aligned along a direction parallel to Bli and B2b so that Si(1) face Ei(l) and Si(mi) face Ei(ni) in a direction parallel to Cli and Dlb - Subunit U2 comprises • a hollowed-out portion in a plate X, possibly in the aforementioned plate X of the aforementioned subunit Ui, • in the shape of a rectangular parallelepiped with 4 lateral faces (a2, b2, c2, d2) and 2 upper and lower faces (e2, f2), • said hollowed-out part being of length Lo2 and width La2 • of height h2 corresponding to the thickness of the plate X-ray electrophoresis, from 25 µm to 20 mm, • the lateral faces (a2, b2) being parallel to each other, the face (a2) being delimited by two edges (Al2, A22) of dimension La2 and the face (b2) being delimited by two edges (Bl2, B22) of dimension La2, • the lateral faces (c2, d2) being parallel to each other, the face (c2) being delimited by two edges (Cl2, C22) of dimension Lo2 and the face (d2) being delimited by two edges (Dl2, D22) of dimension Lo2, • n2 successive entries E2(1), E2(2) to E2(n2-1), E(n2), n2 being an integer from 4 to 9, distributed on the face (a2) between Al2 and A22 and aligned along a direction parallel to Al2 and A22, • m2 successive exits from S2(l), S2(2) to S2(m2-1), S2(m2), m2 being an integer from 4 to 12, distributed on the face (b2) between B12 and B22 and aligned along a direction parallel to B12 and B22, such that S2(l) faces E2(l) and S2(m2) faces E2(n2) along a direction parallel to B12 and B22,

[0224] characterized in that it comprises - an intermediate channel Vb linking the two subunits Ui and U2

[0225] by one of the outputs SVi(Ui) chosen from Si(2) to Si(mrl) of the subunit Ui to one of the inputs EVi(U2) of the subunit U2 chosen from E2(2) to E2(n2-1).

[0226] The cell according to the invention may include 2 subunits Ui and U2 and supply and evacuation channels and an outlet recovery channel in Us.

[0227] According to a particular embodiment, the present invention relates to a cell as defined above, in which 5 equals 2, comprising: - a supply channel configured to connect the Ei(l) inlet of the electrophoresis chamber subunit Ui to an external micro / milli fluidic supply circuit for a first liquid cathode CAL(l), - a supply channel configured to connect the Ei(ni) inlet of the Ui subunit of the electrophoresis chamber to an external micro / milli fluidic supply circuit for a first liquid anode ANL(l), - a supply channel configured to connect at least one of the inputs Ei(2) to Ei(nrl) of the subunit Ui to an external micro / milli fluidic circuit for supplying an initial solution containing a product to be purified and / or separated, - at least one supply channel configured to connect at least one remaining input of the Ui unit to at least one external micro / millifluidic circuit supplying at least one buffer solution, - a supply channel configured to connect the E2(l) inlet of the U2 electrophoresis chamber subunit to an external micro / milli fluidic supply circuit for a second liquid cathode CAL(2), - a supply channel configured to connect the E2(n2) inlet of the U2 subunit of the electrophoresis chamber to an external micro / milli fluidic supply circuit for a second liquid anode ANL(2), - at least one supply channel configured to connect at least one remaining input of unit U2 to at least one external micro / millifluidic circuit supplying at least one buffer solution, - at least one recovery channel configured to connect one of the outputs of the U2 subunit chosen from S2(2) to S2(m2-1) of the electrophoresis chamber to an external micro / milli fluidic recovery circuit,

[0228] said cell being configured for, in the presence of: • of an electric field generated CE(1) in the subunit Ui between the liquid cathode CAL(l) and the liquid anode ANL(l) parallel to Ali and perpendicular to Cli, • of an electric field generated CE(2) in the U2 subunit between the liquid cathode CAL(2) and the liquid anode ANL(2) parallel to Al2 and perpendicular to Cl2, • and in operation,

[0229] a) in the Ui subunit: • circulate the liquid cathode CAL(l) from the inlet Ei(l) to the outlet Si(l), • circulate the liquid anode ANL(l) from the inlet Ei(nO) to the outlet Si(mi), • circulate the initial solution containing the product to be separated and / or purified and at least one buffer solution in the subunit Ui, between the liquid cathode CAL(l) and the liquid anode ANL(l), from the inlets Ei(2) to Ei(nr 1) to the outlets Si(2) to Si(mrl),

[0230] b) in the intermediate channel Vb • recover at the SVi(Ui) output the purified and / or separated product contained in said initial solution; • to circulate the purified and / or separated product from the output SVi(Ui) of subunit Ui through channel Vi, from SVi(Ui) to the input EVi(U2) of subunit U2,

[0231] c) in subunit U2: • circulate the liquid cathode CAL(2) from the inlet E2(l) to the outlet S2(l), • circulate the liquid anode ANL(2) from the inlet E2(n2) to the outlet S2(m2), • circulate said purified and / or separated product from the SVi(Ui) outlet of the Ui subunit and at least one buffer solution in the U2 subunit, between the liquid cathode CAL(2) and the liquid anode ANL(2), from the inlets E2(2) to E2(n2-1) to the outlets S2(2) to S2(m2-1),

[0232] d) recover at one of the outlets S2(2) to S2(m2-1) the purified and / or separated product.

[0233] According to a particular embodiment, the present invention relates to a cell as defined above, in which s is equal to 2, the number of outputs mi of the unit Ui is greater than or equal to 5,

[0234] further comprising an intermediate channel Wi connecting subunit Ui to subunit U2,

[0235] extending: • one of the SWi(Ui) outputs of unit Ui chosen from Si(2) to Si(mrl), different from the SVi(Ui) output of channel Vb • at the EVi(U2) input of the U2 subunit.

[0236] In addition to a portion of the Ui subunit stream collected by the Vi channel and directed to the U2 subunit as a solution to be purified and / or separated, the Wi channel allows another part of the flow collected in the Ui subunit to be directed to complete the solution to be purified and / or separated in the U2 subunit.

[0237] Ratio of subunit dimensions and number of inputs and outputs.

[0238] According to a particular embodiment, the present invention relates to a cell as defined above, in which s is 2, in which the number of outputs mi and m2 of the subunits Ui and U2 is greater respectively than the number of inputs ni and n2 of the subunits Ui and U2.

[0239] According to a particular embodiment, the present invention relates to a cell as defined above, in which 5 is equal to 2 and in which the subunits Ui and U2 have width and length dimensions such that - the width La2 of the subunit U2 is equal to the width Lai of the subunit Ui and - The length Lo2 of the subunit U2 is equal to the length Law of the subunit Up

[0240] According to a particular embodiment, the present invention relates to a cell as defined above, in which 5 is equal to 2 and in which - the width La2 of the subunit U2 is greater than or equal to the width Lai of the subunit Ui and - The length Lo2 of the subunit U2 is less than or equal to the length Law of the subunit Up

[0241] According to a particular embodiment, the present invention relates to a cell as defined above, in which 5 is equal to 2 and in which - the width La2 of the subunit U2 is less than or equal to the width Lai of the subunit Ui and - The length Lo2 of the subunit U2 is greater than or equal to the length Law of the subunit Up

[0242] According to a particular embodiment, the present invention relates to a cell as defined above, in which 5 is equal to 2 and in which - the width La2 of the subunit U2 is less than or equal to the width Lai of the subunit Ui and - The length Lo2 of the subunit U2 is less than or equal to the length Law of the subunit Up

[0243] According to a particular embodiment, the present invention relates to a cell as defined above, in which 5 is equal to 2 and in which - the width La2 of the subunit U2 is greater than or equal to the width Lai of the subunit Ui and - The length Lo2 of the subunit U2 is greater than or equal to the length Law of the subunit Up

[0244] According to a particular embodiment, the present invention relates to a cell as defined above, in which the number of outputs mi and m2 of the subunits Ui and U2 is greater respectively than the number of inputs ni and n2 of the subunits Ui and U2;

[0245] and / or wherein the subunits Ui and U2 have width and length dimensions such that the width La2 of subunit U2 is equal to the width Lai of the subunit Ui and the length Lo2 of subunit U2 is equal to the length Law of subunit Ui,

[0246] or wherein the width La2 of subunit U2 is greater than or equal to the width Lai of subunit Ui and the length Lo2 of subunit U2 is less than or equal to the length Loi of subunit Ub

[0247] or wherein the width La2 of subunit U2 is less than or equal to the width Lai of subunit Ui and the length Lo2 of subunit U2 is greater than or equal to the length Loi of subunit Ub

[0248] or wherein the width La2 of subunit U2 is less than or equal to the width Lai of subunit Ui and the length Lo2 of subunit U2 is less than or equal to the length Loi of subunit Ub

[0249] or wherein the width La2 of subunit U2 is greater than or equal to the width Lai of subunit Ui and the length Lo2 of subunit U2 is greater than or equal to the length Loi of subunit Up

[0250] Electrophoresis cell with one chamber of 3 subunits.

[0251] According to a particular embodiment, the present invention relates to a cell as defined above, in which 5 is equal to 3, the electrophoresis chamber comprising three subunits Ub, U2, and U3, - the subunit Ui comprising • a hollowed-out section in a plate X • in the shape of a rectangular parallelepiped with 4 lateral faces (ab bb Ci, di) and 2 upper and lower faces (eb fi), • said hollowed-out part being of length Loi and width Lai • of height hi corresponding to the thickness of the plate X-ray electrophoresis, from 25 µm to 20 mm, • the lateral faces (ab bj) being parallel to each other, the face (aj) being delimited by two edges (A1bA2J) of dimension Lai and the face (bi) being delimited by two edges (Bli, B2i) of dimension Lap • the lateral faces (cp di) being parallel to each other, the face (ci) being delimited by two edges (Clp C2i) of dimension Law and the face (dj) being delimited by two edges (Dlp D2i) of dimension Law, • neither successive entries Ei(l), Ei(2) to Ei(nrl), Ei(ni), nor being an integer from 4 to 9, distributed on the face (ai) between Ali and A2i and aligned along a direction parallel to Ali and A2p • mi successive exits from Si(1), Si(2) to Si(mi-1), Si(mi), where mi is an integer from 4 to 12, distributed on the face (bi) between Bh and B2i and aligned along a direction parallel to B1 and B2H so that Si(1) faces Ei(1) and Si(mi) faces Ei(ni) along a direction parallel to C1 and D1 The U2 subunit comprises • a hollowed-out section in a plate X • in the shape of a rectangular parallelepiped with 4 lateral faces (a2, b2, c2, d2) and 2 upper and lower faces (e2, f2), • said hollowed-out part being of length Lo2 and width La2 • of height h2 corresponding to the thickness of the plate X-ray electrophoresis, from 25 µm to 20 mm, • the lateral faces (a2, b2) being parallel to each other, the face (a2) being delimited by two edges (Al2, A22) of dimension La2 and the face (b2) being delimited by two edges (Bl2, B22) of dimension La2, • the lateral faces (c2, d2) being parallel to each other, the face (c2) being delimited by two edges (Cl2, C22) of dimension Lo2 and the face (d2) being delimited by two edges (Dl2, D22) of dimension Lo2, • n2 successive entries E2(1), E2(2) to E2(n2-1), E(n2), n2 being an integer from 4 to 9, distributed on the face (a2) between Al2 and A22 and aligned along a direction parallel to Al2 and A22, • m2 successive exits from S2(l), S2(2) to S2(m2-1), S2(m2), m2 being an integer from 4 to 12, distributed on the face (b2) between B12 and B22 and aligned along a direction parallel to Bl2 and B22, so that S2(l) faces E2(l) and S2(m2) faces E2(n2) along a direction parallel to Cl2 and Dl2, The U3 subunit comprises • a hollowed-out section in a plate X • in the shape of a rectangular parallelepiped with 4 lateral faces (a3, b3, c3, d3) and 2 upper and lower faces (e3, f3), • said hollowed-out part being of length Lo3 and width La3 • of height h3 corresponding to the thickness of the plate X-ray electrophoresis, from 25 µm to 20 mm, • the lateral faces (a3, b3) being parallel to each other, the face (a3) ​​being delimited by two edges (Al3, A23) of dimension La3 and the face (b3) being delimited by two edges (Bl3, B23) of dimension La3, • the lateral faces (c3, d3) being parallel to each other, the face (c3) being delimited by two edges (Cl3, C23) of dimension Lo3 and the face (d3) being delimited by two edges (Dl3, D23) of dimension Lo3, • n3 successive entries E3(1), E3(2) to E3(n3-1), E(n3), n3 being an integer from 4 to 9, distributed on the face (a3) ​​between Al3 and A23 and aligned along a direction parallel to Al3 and A23, • m3 successive exits from S3(l), S3(2) to S3(m3-1), S3(m3), m3 being an integer from 4 to 12, distributed on the face (b3) between B13 and B23 and aligned along a direction parallel to B13 and B23, such that S3(l) faces E3(l) and S3(m3) faces E3(n3) along a direction parallel to B13 and B23,

[0252] characterized in that it comprises - an intermediate channel Vj, connecting the two subunits Ui and U2

[0253] by one of the outputs SVi(Ui) chosen from Si(2) to Si(mrl) of the subunit Ui to one of the inputs EVi(U2) of the subunit U2 chosen from E2(2) to E2(n2-1), and - an intermediate channel V2, linking the two subunits U2 and U3

[0254] by one of the outputs SV2(U2) chosen from S2(2) to S2(m2-1) of the subunit U2 to one of the inputs EV2(U3) of the subunit U3 chosen from E3(2) to E3(n3-1).

[0255] Electrophoresis device

[0256] Another object of the invention relates to a free-flow electrophoresis cell device comprising: - p plates X between two adjacent plates Y: • p being an integer from 1 to 50, in particular from 1 to 20, • X being an electrophoresis plate (1) made of inert material, • Y being a sealed plate (2) of sapphire or of alumina A12O3 with 99% a-Al2O3, - f electrophoresis cells as defined above, where f is an integer from 1 to 500, - means of clamping all the plates allowing the sealing of said device.

[0257] The device of the invention is an arrangement of one to several hundred electrophoresis cells, in particular from 1 to 500 electrophoresis cells, the cells being as defined above. The electrophoresis cell forms a constituent and repeatable unit of the device.

[0258] The use of the sapphire Y-plate provides a robust and watertight device that allows for disassembly and reassembly, facilitating the cleaning of the device's components and its maintenance. The modularity of the device allows for the reuse of the plates.

[0259] Advantageously, the mechanical strength of sapphire allows for an assembly system that enables significant pressure from the flows in the device while ensuring excellent sealing between each stage, which is not the case for glass, which scratches and cracks very easily under slight pressure; in the presence of water, the crack in the glass can propagate throughout the cell and induce significant sealing problems.

[0260] Thus, the use of sapphire Y-plates as adjacent plates closing a fluidic circuit allows for robustness and temperature control in the device thanks to the thermal conductivity of sapphire. Consequently, it provides a free-flow electrophoresis device capable of processing industrial volumes, for industrial applications of continuous sample purification and / or separation by free-flow electrophoresis.

[0261] Subunits shared by two electrophoresis chambers.

[0262] According to a particular embodiment, the present invention relates to a device as defined above, comprising at least two electrophoresis cells such that in at least one of their subunit Ut, t being an integer from 1 to (s-1), at least two of their respective outputs lead to the same input of the same successive subunit Ut+i through their respective channel Vt.

[0263] In this embodiment, for at least two successive subunits, respectively named the first and second subunits, the second subunit has an inlet linked to two intermediate channels V originating from two different first subunits. Thus, a second subunit is distributed by two distinct first subunits, thereby increasing the flow rate of the solution to be purified and / or separated.

[0264] According to a particular embodiment, the present invention relates to a device as defined above, comprising at least one pair of two electrophoresis cells G1 and G2 of identical configuration and each comprising two subunits,

[0265] in which - Vi(Gl) is the intermediate Vi channel of cell G1 connecting the subunit Ui of Gl, named Ui(Gl), to the subunit U2 of G1 named U2(G1), from the output SVi(Ui(Gl)) to the input EVi(U2(G1)), - Vi(G2) is the intermediate Vi channel of cell G2 connecting the subunit Ui of G2, named Ui(Gl), to the subunit U2 of Gl named U2(G1), from the output SVi(Ui(G2)) to the input EVi(U2(G2)),

[0266] and wherein the aforementioned Vi(Gl) and Vi(G2) channels open onto the same input of the U2 subunit.

[0267] It is understood that the aforementioned entries EVi(U2(G1)) and EVi(U2(G2)) represent the same entry, named EVi(U2(G1-G2)) and the aforementioned subunits U2(G1) and U2(G2) represent the same subunit, named U2(G1-G2).

[0268] The arrangement of the two electrophoresis chambers and their channels Vi(G1) and Vi(G2) is configured so as to distribute the purified and / or separated product from Ui(G1) through the Vi(G1) channel and the purified and / or separated product from Ui(G2) through the Vi(G2) channel to the same subunit U2(G1-G2) via the same inlet EVi(U2(G1-G2)) of the aforementioned subunit EVi(U2(G1-G2)). This allows the flow rate of the solution to be purified and / or separated in the second subunit, which is common to both electrophoresis chambers, to be doubled.

[0269] According to a particular embodiment, the present invention relates to a device as defined above, in which

[0270] each plate named Xj, j being an integer ranging from 1 to p, comprises q, electrophoresis cells as defined above, q, being an integer from 1 to 10.

[0271] In this embodiment, the device comprises p plates X, named Xi to Xp, p being an integer from 1 to 50 and each plate comprises from 1 to 10 electrophoresis cells, the number of electrophoresis cells on plate Xj being named q. The sum of the number of cells in each plate q, when [ ] varies from 1 to p is equal to the total number f of cells in the device.

[0272] When the subunits of the electrophoresis chamber extend over several plates X due to the use of distinct heights of the hollowed parts, the number q of electrophoresis cells on the plate Xj corresponds to the number of subunits Ui on the plate Xj.

[0273] Chamber subunits on the same plate

[0274] According to a particular embodiment, the present invention relates to a device as defined above,

[0275] in which - p is an integer from 1 to 50, in particular from 1 to 10, - when j varies from 1 to p, q is an integer from 2 to 50, in particular from 2 to 10

[0276] comprising p electrophoresis plates Xj each having from 2 to 50 electrophoresis chambers, in particular from 2 to 10 electrophoresis chambers, each electrophoresis chamber comprising 5 subunits, 5 representing an integer from 2 to 5,

[0277] wherein for each electrophoresis chamber, the hollowed-out parts of the 5 subunits are located in the same plate.

[0278] In this embodiment, the subunits Ui to Us of the electrophoresis chamber are located on the same plate X. The device comprises p plates X, named Xi to Xp, p being an integer from 1 to 50 and each plate comprises from 1 to 50 electrophoresis cells with s subunits, s from 2 to 5.

[0279] Advantageously the X plates all comprise the same number of electrophoresis cells.

[0280] Electrophoresis chambers on the same plate

[0281] According to a particular embodiment, the present invention relates to a device as defined above, in which p is equal to 1 and qi is an integer from 2 to 50, in particular from 2 to 10, comprising a single electrophoresis plate Xi having from 2 to 50 electrophoresis chambers, in particular from 2 to 10 chambers, each electrophoresis chamber comprising 5 subunits, 5 representing an integer from 2 to 5.

[0282] In this embodiment, the device comprises a single plate X which has from 2 to 50 electrophoresis chambers, in particular from 2 to 10 electrophoresis chambers, each chamber containing s subunits, s representing an integer from 2 to 5.

[0283] Row alignment of subunits

[0284] According to a particular embodiment, the present invention relates to a device as defined above, in which in each plate Xj, the q, electrophoresis chambers are aligned so that the subunits U; of the q, electrophoresis chambers are adjacent to each other by the faces (c;) or (d;).

[0285] In this embodiment, the U subunits of the same index i in each electrophoresis chamber of the same plate are aligned in a row. Thus, the Ui subunits of the electrophoresis chambers form one row, and the U2 subunits form another row. This arrangement advantageously optimizes the cooling of the electrophoresis chambers by the cooling circuit and the use of the supply, recovery, and discharge channels of the device.

[0286] According to a particular embodiment, the present invention relates to a device as defined above, in which - p is an integer from 1 to 50, in particular from 1 to 10, - when j varies from 1 to p, q is an integer from 2 to 50, in particular from 2 to 10,

[0287] comprising p electrophoresis plates Xj each having from 2 to 50 electrophoresis chambers, in particular from 2 to 10 electrophoresis chambers, each electrophoresis chamber comprising 5 subunits, 5 representing an integer from 2 to 5,

[0288] wherein for each electrophoresis chamber, the hollowed-out parts of the 5 subunits are located in the same plate.

[0289] or in which

[0290] p is equal to 1 and qi is an integer from 2 to 50, in particular from 2 to 10, comprising a single electrophoresis plate Xi having from 2 to 50 electrophoresis chambers, in particular from 2 to 10 electrophoresis chambers, each electrophoresis chamber comprising 5 subunits, 5 representing an integer from 2 to 5,

[0291] and / or wherein in each plate Xj, the q, electrophoresis chambers are aligned so that the subunits U; of the q, electrophoresis chambers are adjacent to each other by the (cO or (d;) faces.

[0292] According to a particular embodiment, the present invention relates to a device as defined above, comprising a supply channel network configured to connect at least one of the inputs Ei(2) to Ei(nrl) of the subunit Ui of each electrophoresis chamber to an external micro / milli fluidic circuit for supplying an initial solution containing a product to be purified and / or separated.

[0293] In this embodiment, the supply of initial solution to be purified and / or separated in the electrophoresis chambers is carried out in parallel.

[0294] Device with cooling plate.

[0295] According to a particular embodiment, the present invention relates to a device as defined above, comprising cooling means.

[0296] According to a particular embodiment, the present invention relates to a device as defined above, comprising a vertical succession of plates X, Y, Z whose surfaces are stacked according to the sequence YZY(XYZY)p,

[0297] in which

[0298] X represents an electrophoresis plate made of inert material,

[0299] Y represents a sealed plate made of sapphire or alumina (Al2O3) with 99% a-Al2O3,

[0300] Z represents a cooling plate comprising a heat transfer system (4),

[0301] p, an integer from 1 to 50, in particular from 1 to 20, represents both the number of stages of said device and the number of plates X

[0302] each floor being defined:

[0303] - by the following sequence of plates YZYXYZY, in which:

[0304] - plate X is located between two plates Y,

[0305] - each of the two plates Z being respectively adjacent to a plate Y,

[0306] - and each of the two Y plates located at the ends of the YZYXYZY sequence, covers a plate Z respectively so that each plate Z is located between two plates Y.

[0307] By "vertical succession of plates" is meant a stacking of plates, in which the surfaces of the different plates are in contact.

[0308] Device with membrane

[0309] According to a particular embodiment, the present invention relates to a device as defined above, in which each electrophoresis chamber is configured to contain at least one membrane with selective permeability, in particular selective in size, positioned so as to be traversed by the solution containing the product to be separated or purified during the operation of the device,

[0310] preferably said at least one selective permeability membrane is placed in the Us subunit of each electrophoresis chamber.

[0311] The presence of these membranes also allows for selectivity of sizes of the product to be purified and / or separated.

[0312] Advantageously the membrane can be positioned in a subunit U; parallel to the face (Cj) and adjacent to an inlet of a buffer solution, so as to be crossed by the initial solution or by the solution to be purified and / or separated during the operation of the device and so that the part of said solution not having crossed said membrane is conveyed to one of the outlets by a buffer solution from an inlet adjacent to said membrane.

[0313] Device with protrusion.

[0314] According to a particular embodiment, the present invention relates to a device as defined above, in which the upper faces e; and / or the lower faces fi of the subunits U; of each of the electrophoresis chambers comprise at least one protrusion configured so as not to disturb, during the operation of the device, the circulation in the chamber of the product to be purified and / or separated, and configured to improve heat transfer and to maintain each of said electrophoresis chambers at a selected temperature,

[0315] in particular wherein said at least one protuberance is made of thermally conductive material, preferably sapphire or 99% alumina a-Al2O3.

[0316] The protrusions extend from faces (e) and / or (f) of the hollowed portion towards the interior of said hollowed portion. They may be located only on one of the faces or on both faces.

[0317] The presence of these protrusions promotes heat exchange between the cooling plates along the path and thus optimizes the separation of the desired molecule while avoiding denaturation of the molecules, while maintaining the optimal temperature.

[0318] According to a particular embodiment, the present invention relates to a device as defined above, in which each electrophoresis chamber is configured to contain at least one selective permeability membrane, in particular size-selective, positioned so as to be traversed by the solution containing the product to be separated or purified during the operation of the device, preferably said at least one selective permeability membrane is placed in the Us subunit of each electrophoresis chamber;

[0319] and / or wherein the upper faces e; and / or the lower faces £ of the subunits U; of each of the electrophoresis chambers comprise at least one protrusion configured not to disturb, during operation of the device, the circulation in the chamber of the product to be purified and / or separated, and configured to improve heat transfer and to maintain each of said electrophoresis chambers at a selected temperature,

[0320] in particular wherein said at least one protuberance is made of thermally conductive material, preferably sapphire or 99% alumina a-Al2O3;

[0321] and / or said device includes a supply channel network configured to connect less one of the inputs Ei(2) to Ei(nrl) of the subunit Ui of each electrophoresis chamber to an external micro / milli fluidic supply circuit of an initial solution containing a product to be purified and / or separated.

[0322] Usage

[0323] Another object of the present invention relates to the use of an electrophoresis cell according to the invention as defined above or of an electrophoresis cell device according to the invention as defined above, in the implementation of a process of purification and / or separation by free flow electrophoresis, of a product to be purified and / or separated contained in an initial solution, comprising a first step of purification and / or separation in the subunit(s) Ui followed by (s-1) successive polishing step(s) in the subunits U2 to Us of the electrophoresis chamber(s).

[0324] Electrophoresis purification and / or separation process

[0325] Another object of the present invention relates to a method for purifying and / or separating, by free-flow electrophoresis, a product to be purified and / or separated contained in an initial solution by implementing an electrophoresis cell device as defined above, comprising the following steps:

[0326] a) a purification and / or separation step of the aforementioned product in the Ui subunits of each electrophoresis chamber, under an electric field CE(1), comprising:

[0327] in the Ui subunit of each electrophoresis chamber: • the circulation of a liquid cathode CAL(l) from the inlet Ei(l) to the outlet Si (1), • the circulation of a liquid anode ANL(l) from the inlet E|(nJ to the outlet Si(mi), • the circulation of the initial solution containing the product to be separated and / or purified and at least one buffer solution in the Ub subunit between the liquid cathode CAL(l) and the liquid anode ANL(l), from the inlets Ei(2) to Ei(nr 1) to the outlets Si(2) to Si(mi-l), to obtain the purified and / or separated product from the Ui subunit,

[0328] b) (s-1) polishing step(s) k, k varying from 1 to (s-1), each step k comprising each:

[0329] i) in the intermediate channel Vk of each electrophoresis chamber, • the recovery at the SVk(Uk) outlet corresponding to the intermediate channel Vk of said purified and / or separated product from the Uk subunit; • the flow, in the absence of an electric field, of the aforementioned purified and / or separated product, from the output SVk(Uk) of the subunit Uk corresponding to the intermediate channel Vk to the input EVk(Uk+i) of the subunit Uk+i corresponding to the intermediate channel Vk,

[0330] ii) in the Uk+i subunit of each electrophoresis chamber, in the presence of an electric field CE(k+l): • the circulation of a liquid cathode CAL(k+l) from the inlet Ek+[(1) to the outlet Sk+1(l), • the circulation of a liquid anode ANL(k+l) from the inlet Ek+i(nk+i) to the outlet Sk+i(mk+i), • the circulation of said purified and / or separated product from the outlet of the Uk subunit and at least one buffer solution in the Uk+i subunit, between the liquid cathode CAL(k+l) and the liquid anode ANL(k+l), from the inlets Ek+[(2) to Ek+1(nk+1-l) to the outlets Sk+i(2) to Sk+i(mk+1-l),

[0331] to obtain the product (k+1) once purified and / or separated from the Uk+i subunit;

[0332] c) a recovery step comprising: - the recovery at one of the outputs Ss(2) to Ss(ms-1) of the unit Us of each electrophoresis chamber of the product(s) times purified and / or separated.

[0333] Electric fields

[0334] According to a particular embodiment, the present invention relates to a process as defined above, in which each electric field CE(i), generated in the subunit U;, i an integer varying from 1 to s, during the fluidic circulation in each of the electrophoresis chambers, is from 200V to 4000V.

[0335] According to a particular embodiment, the present invention relates to a method as defined above, in which at least two values ​​of electric fields CE(i), generated respectively in the subunits Ui, i an integer varying from 1 to s, are different.

[0336] According to a particular embodiment, the present invention relates to a process as defined above, in which said process is carried out under continuous flow of the initial solution containing the product to be purified and / or separated.

[0337] According to a particular embodiment, the present invention relates to a method as defined above, implemented by a device as defined above, in which s is equal to 2, comprising the following steps:

[0338] a) a purification and / or separation step of the product to be purified and / or separated in the Ui subunits of each electrophoresis chamber, under an electric field CE(1), comprising:

[0339] in the Ui subunit of each electrophoresis chamber: • the circulation of a liquid cathode CAL(l) from the inlet Ei(l) to the outlet Si (1), • the flow of a liquid anode ANL(l) from the inlet Ei(ni) to the outlet Si(mi), • the circulation of the initial solution containing the product to be separated and / or purified and at least one buffer solution in the Ui subunit, between the liquid cathode CAL(l) and the liquid anode ANL(l), from the inlets Ei(2) to Ei(nr 1) to the outlets Si(2) to Si(mi-l), to obtain the purified and / or separated product from the Ub subunit

[0340] b) a polishing step comprising:

[0341] i) in the intermediate channel, empty each electrophoresis chamber, • the recovery at the SVi(Ui) output corresponding to the intermediate channel Vi of said purified and / or separated product from the Ui subunit; • the flow, in the absence of an electric field, of the aforementioned purified and / or separated product, from the output SVi(Ui) of the subunit Ui corresponding to the intermediate channel Vi to the input EVi(U2) of the subunit U2 corresponding to the intermediate channel Vi,

[0342] ii) in the U2 subunit of each electrophoresis chamber, in the presence of an electric field CE(2): • the circulation of a liquid cathode CAL(2) from the inlet E2(l) to the outlet S2 (1), • the circulation of a liquid anode ANL(2) from the inlet E2(n2) to the outlet S2(m 2), • the circulation of said purified and / or separated product from the outlet of subunit U2 and at least one buffer solution in subunit U2, between the liquid cathode CAL(2) and the liquid anode ANL(2), from the inlets E2(2) to E2(n2-1) to the outlets S2(2) to S2(m2-1),

[0343] to obtain the purified and / or separated and polished product from the U2 subunit;

[0344] c) a recovery step comprising: - the recovery at one of the outlets S2(2) to S2(m2-1) of the U2 unit of each electrophoresis chamber of the purified and / or separated product, once polished.

[0345] Figures and examples

[0346] Fig. 1 represents the hollowed-out portion of a subunit of an electrophoresis chamber of an electrophoresis plate X (1) between two adjacent plates Y, respectively upper and lower. The hollowed-out portion is inscribed within a rectangular parallelepiped of width La, length Lo, and height h, delimited by the faces (ai, bi, ci, di). The faces (ai, bi, ci, di) form the lateral walls between the hollowed-out portion and the plate X, the faces (ai, bi) being parallel to each other and the faces (ci, di) being parallel to each other. The face (ai) is delimited by the edges (Ah, A2) of dimension La, the face (bi) by the edges (Bh, B2) of dimension La. The face (cO) is delimited by the edges (CL, C2;) of dimension Lo;, the face (d;) by the edges (DL, D2; ) of dimension Lo;. The edges (AL, BL, CL, DL) delimit the face (e;) formed by the upper plate Y and the edges (A2;, B2;, C2;, D2;) delimit the face (L) formed by the lower plate Y.

[0347] Figure 2 shows a schematic diagram of an electrophoresis chamber comprising two subunits. The first subunit Ui comprises 9 inputs from Ei(1) to Ei(9) and 5 outputs Si(1) to Si(5), with input Ei(1) facing output Si(1) and input Ei(9) facing output Si(5). The second subunit U2 comprises 8 inputs from E2(1) to E2(8) and 12 outputs from S2(1) to S2(12), with input E2(1) facing output S2(1) and input E2(8) facing output S2(12). The subunit Ui is connected to the second subunit U2 by an intermediate channel Vj which extends from the output Si(4) of Ui, named SVi(Ui) to the input E2(4) of U2, named EVi(U2).

[0348] Figure 3 shows a schematic of an electrophoresis chamber comprising three subunits. The subunit U1 is connected by an intermediate channel V1 to the subunit U2. The subunit U2 is connected by an intermediate channel V2 to the subunit U3.

[0349] Figure 4 shows in part (a) an exploded view diagram of an electrophoresis cell comprising a plate X between two adjacent plates Y forming a YXY sequence. Plate X comprises a chamber of two subunits Ui and U2 connected by an intermediate channel Vi. Subunit Ui has 7 inputs from Ei(1) to Ei(7) and 5 outputs from Si(1) to Si(5), with the output of the intermediate channel SVi(Ui) corresponding to Si(3). Subunit U2 has 7 inputs from E2(1) to E2(7) and 7 outputs from S2(1) to S2(5), with the input of the intermediate channel EVi(U2) corresponding to E2(4). The upper Y plate and the X plate are engraved the supply and retrieval channels of the outputs Si(1), Si(2), Si(4) and Si(5) and of the inputs E2(1), E2(2), E2(3), E2(5), E2(6), E2(7), the output Si(3) and the input E2(4) corresponding to the intermediate channel Vj being excluded.

[0350] In part b), (1) represents an electrophoresis plate comprising a single chamber having two rectangular parallelepiped-shaped subunits (6), including inlets or outlets (7) and supply or return channels (8). (2) represents a synthetic sapphire plate ensuring the separation of fluids between two plates but allowing heat exchange. (3) represents a cooling plate Z comprising a cooling system (4) which includes a recess allowing the circulation of a heat transfer fluid from an inlet to an outlet, the inlet of the heat transfer fluid being on the same side as the inlets of the electrophoresis chamber. The device consists of a succession of plates YZYXYZY.

[0351] Fig. 5 represents the schematic of an electrophoresis chamber comprising two subunits and including a complementary intermediate channel W, in part a) the intermediate channel Vj and the complementary intermediate channel W are adjacent, in part b) the two channels are not adjacent, an outlet between the two channels allows the recovery of a compound from the product to be purified and / or separated.

[0352] Fig. 6 represents the two-subunit electrophoresis chamber scheme, in which two Ui subunits distribute the same U2 subunit; in part a), the respective outputs of the two intermediate channels Vi correspond to different outputs of the Ui(1) and Ui(2) subunits; in part b) the respective outputs of the two intermediate channels Vi correspond to the same subunit output.

[0353] Figure 7 shows the schematic of a plate X of a free-flow electrophoresis device comprising 10 electrophoresis chambers of two subunits of Figure 2. The 10 U1 subunits form one row and the U2 subunits form another row.

[0354] Figure 8 shows two devices in exploded view and without representation of the clamping means for all the plates, in part a) a single-stage device comprising a row of 10 electrophoresis chambers and in part b) a two-stage device, each stage comprising a row of 10 electrophoresis chambers.

[0355] (1) represents an electrophoresis plate comprising a row of 10 chambers of electrophoresis, each comprising two rectangular parallelepiped-shaped subunits (6), each chamber including inlets or outlets and supply or recovery channels; two adjacent subunits are separated by a single wall. (2) represents a synthetic sapphire plate ensuring the separation of fluids between two plates but allowing heat exchange. (3) represents a cooling plate comprising a cooling system (4) which includes a recess allowing the circulation of a heat transfer fluid from an inlet to an outlet. (5) represents a stage consisting of a succession of plates YZYXYZY.

[0356] The device in part a) comprises 1 stage and consists of the following sequence YZYXYZY.

[0357] The device in part b) comprises 2 stages and consists of the following sequence YZYXYZYXYZY in which the central sequence YZY is common to both stages.

[0358] Figure 9 shows a diagram of a device with a plate clamping system. In this particular embodiment, the clamping means consist of two plates (11) that clamp the entire sequence of plates X, Y, and Z, by means of fastening means (12) connecting the two plates (11), the distance between which can be adjusted. The fastening means (12) are, for example, screws.

[0359] Fig. 10 represents the schematic of the system used for the implementation to evaluate the purification and separation yields of a subunit.

[0360] Fig. 11 represents the schematic of a chamber with two identical subunits used for implementation to evaluate the purification and separation yield of an initial solution consisting of 3 dyes (fluorescein, rhodamine B and rhodamine 6G).

[0361] The [Fig. 12] is a photograph taken of the Ui subunit of an electrophoresis chamber having 5 inlets and 9 outlets, with a thickness of 100 pm, during a separation of a mixture of three colored compounds (fluorescein, rhodamine B and rhodamine 6G), carried out at 2500 V.

[0362] Fig. 13 represents the HPLC spectra of the products at the outputs of the first subunit Ui of an electrophoresis chamber having 5 inlets and 9 outlets, with a thickness of 100 pm, during a separation test of a mixture of three colored compounds (fluorescein, rhodamine B and rhodamine 6G), at 0V and at 2500 V.

[0363] Figure 14 is a graph representing the output flow rate of each dye at the different outlets of the first subunit Ui of a chamber electrophoresis apparatus comprising 5 inlets and 9 outlets, with a thickness of 100 pm, during a separation test of a mixture of three colored compounds (fluorescein, rhodamine B and rhodamine 6G), at 2500 V.

[0364] Fig. 15 represents the HPLC spectra of the products at the outputs of the second U2 subunit of an electrophoresis chamber having 5 inlets and 9 outlets, with a thickness of 100 pm, during a separation test of a mixture of three colored compounds (fluorescein, rhodamine B and rhodamine 6G), at 0V and at 2500V.

[0365] The [Fig. 16] is a graph representing the output flow rate of each dye at the different outputs of the second U2 subunit of an electrophoresis chamber having 5 inlets and 9 outlets, with a thickness of 100 pm, during a test of separation of a mixture of three colored compounds (fluorescein, rhodamine B and rhodamine 6G), at 2500 V.

[0366] Example 1: Optimization of the separation of a mixture of Fluorescein, Rhodamine B and Rhodamine 6G in a two-subunit electrophoresis cell. Materials and methods

[0367] The electrophoresis system

[0368] The electrophoresis system consists of an air supply system that feeds a pump connected to flow meters controlling the flow inside the electrophoresis chip ([Fig. 10]). This system ensures a stable flow rate for stable operation. The electrophoretic chip is interchangeable depending on the separation requirements of the various media. Before each experiment, the system requires a start-up and a stabilization period to ensure steady-state operation (stabilized flow rates, pressure, and temperature).

[0369] Model molecules

[0370] The free-flow electrophoresis system was set up for the separation of a mixture of three fluorescent molecules: fluorescein, rhodamine B, and rhodamine 6G. These are small dyes with similar molar masses, each with a distinct color that allows for visual identification prior to HPLC analysis. This visual check also allows for verification of cell operation before application of the electric field.

[0371] These dyes are easily analyzed by HPLC, they have distinct retention times, and their calibration curves have been defined. By HPLC, the retention time of: • Fluorescein, is 5.9 minutes, • Rhodamine B, is 7.6 minutes • Rhodamine 6G, is 8.6 minutes.

[0372] The chemical structures of the three molecules are shown below. Rhodamine SG Rhodamine B Fluorescein Mm = 479.01 g / mo! (Purple) Mm= 479.01 g / me (Red) Mm = 332.3 g / mol (Yellow) QsHîiCINjOb QsHsiClhbCF CzoHîzOs

[0373] These dyes also possess, in aqueous solution, a different surface charge that allows their migration and separation under the application of an electric field. This surface charge was measured by analyzing the zeta potential with the Malvern Zetasizer. The results obtained (Table 1) show that the three dyes have a significant difference in zeta potential and therefore in surface charge. The zeta potential of each compound was measured beforehand and is reported in Table 1.

[0374] Fluorescein at pH 7.49 exhibited a zeta potential of -23.5 mV, rhodamine B -0.3 mV and rhodamine 6G +9.4 mV.

[0375] [Tables 1] Compound Measured Zeta Potential (mV) at pH 7.5 Average Zeta Potential at pH 7.5 Fluorescein -27.85 -21.05 -21.06 -23.5 mV Rhodamine B -0.6 -0.1 -0.3 -0.3 mV Rhodamine 6G +10.62 +9.26 +8.41 +9.429 mV

[0376] Table 1: Measured zeta potential of the three dyes

[0377] It is important to note that rhodamine B is weakly charged and therefore its electrophoretic mobility is very low regardless of the strength of the electric field.

[0378] The separation and purification of a sample containing a mixture of these 3 compounds, in particular containing rhodamines B and 6G, was carried out in each of the two subunits constituting the electrophoresis cell, by analyzing the output fluxes in order to demonstrate:

[0379] - the technological feasibility of the cell composed of 2 or even more subunits who cooperate.

[0380] - the scientific interest relating to the continuous separation of various compounds with similar properties such as the mass of biomolecules.

[0381] The following examples describe the tests set up to separate the mixture of the 3 dyes in a first subunit U1 followed by a separation polishing of a sample stream exiting U1 introduced into a subunit U2, as illustrated in [Fig. 11]. The sample recovered in the second subunit corresponds to the output comprising a Rhodamine B / Rhodamine 6G mixture obtained at the outlet of the first subunit after the first separation step with a 9-output subunit. The liquid electrodes and the separation buffer maintain the same compositions in both subunits.

[0382] Example 2: First electrophoretic separation step in the first Ui subunit

[0383] A separation of a sample consisting of a mixture of the 3 dyes (fluorescein, rhodamine B and rhodamine 6G) was set up with a Ui subunit of EAD-008 configuration which comprises:

[0384] - ni = 5 entries

[0385] - mi = 9 outputs,

[0386] - a height hi of 100 pm

[0387] - width Lai of 3 cm,

[0388] - Length Law of 6.4 cm

[0389] The flow rates of the flows injected using the flow meters into the inlets of subunit Ui are, for the sample, buffer solutions and liquid electrode, respectively, 10 / 100 / 20 pL / min. The inlet solution setup is shown in Table 2 below.

[0390] The applied electric field voltage CE(1) is fixed at 2500 V.

[0391] [Tables2] Inlet Injected Solution Composition Flow Rate (pL / min) ^(1) Liquid Electrode (Cathode) Water / methanol (70 / 30), HEPES 10mM, HPMC 0.2%(w / v), KCl 1.5M pH = 7.75 Conductivity 102.6 mS / cm 20 Viscosity (25°C) 2.19 mPa / s E^) Buffer solution Water HEPES at 10mM, HPMC at 0.2%(w / v), Tween 20 at 0.1%(w / v) pH = 7.61 Conductivity 393.6 mS / cm Viscosity (25°C) 1.22 mPa / s 100 Ei(3) Sample = initial solution Mixture of 3 dyes: -Fluorescein, -Rhodamine 6G and -Rhodamine B Water 10 Ei(4) Buffer solution Water HEPES at 10mM, HPMC at 0.2%(w / v), Tween 20 at 0.1%(w / v) pH = 7.61 Conductivity 393.6 mS / cm Viscosity (25°C) 1.22 mPa / s 100 Ei(5) Liquid electrode (anode): Water / methanol (70 / 30), 10 mM HEPES, 0.2% (w / v) HPMC, 1.5 M KCl, pH = 7.75, Conductivity 102.6 mS / cm, Viscosity (25°C) 2.19 mPa / s

[0392] Table 2: Operating conditions in the inputs Ei(1) to Ei(5) of the subunit Ui

[0393] Thus, the inputs Ei(l) and Ei(5) were each supplied by a solution The electrolytic solutions for the anode and cathode are respectively identical. The liquid electrolytic solution has the following composition: HEPES at 10mM, HPMC 0.2% (w / v) electrolyte solution, 40% methanol, and 1.5M KCl were used. The flow rate was set at 20 pL / min. These electrolytic solutions exhibited a pH of 7.75, a conductivity of 102.6 mS / cm, and a viscosity of 2.19 mPa / s at 25°C. The electric field was generated by the electrolytic solutions in the electrophoresis chamber, which were prepared from solutions containing carbon electrodes, anode, and cathode, respectively.

[0394] The central inlet Ei(3) was fed with the sample to be purified, composed of 0.156 mg / mL of fluorescein, 0.23 mg / mL of Rhodamine 6G and 0.25 mg / mL of Rhodamine B; The flow rate set for the sample was 10 pL / min throughout the purification.

[0395] The two inlets Ei(2) and Ei(4) were supplied with a buffer solution composed of HEPES at 10 mM, HPMC at 0.2% (nVv), Tween 20 at 0.1% (m / v) in water, at a fixed flow rate of 100 pL / min. The buffer solution had a pH of 7.61, a conductivity of 393.6 pS / cm, and a viscosity at 25°C of 2.19 mPa / s.

[0396] The electric field was set at 2500 V. After stabilization of the flows, i.e. the system was left running for 1h45 minutes and the products at the outlet of the electrophoresis chamber were collected in tubes and analyzed by HPLC in order to determine the percentage of each compound at each outlet.

[0397] A control test was carried out with an electric field fixed at 0 V. The system was left running continuously for 30 minutes.

[0398] Example 3 - Result of electrophoretic separation in the Ui subunit Visual analysis

[0399] The [Fig. 12] is a photograph taken of the Ui subunit under an electric field of 2500 V during electrophoresis made possible by the use of transparent sapphire plates and colored products to be separated.

[0400] At 2500 V, [Fig. 12] visually shows an initial sample beam separating into 3 beams of different colors, mainly centered respectively at the output Si(3) of yellow color, at the output Si(5) of violet color, and at the output Si(6) of red color.

[0401] At 0 V, the sample beam composed of the 3 dyes is visible and centered. There is no electric field, so no deflection is visible. The dyes exit only at the Si(5) output, which is the central output facing the Ei(3) input. HPLC Analysis

[0402] At 2500 V, the outputs of the Si(1) to Si(9) electrophoresis subunit were analyzed by HPLC.

[0403] For the control, at 0 V, only the outputs of the electrophoresis subunit Si(4) to Si(6) were analyzed.

[0404] The HPLC spectra of the products exiting at the 0V and 2500V outputs are shown in [Fig.13].

[0405] The relative distribution of the concentration of each dye in the outputs by HPLC analysis at 2500 V is reported in Table 3.

[0406] [Tables3] Si(D Si(2) Si(3) Si(4) Si(5) Si(6) Si(7) Si(8) Si(9) Fluorescein (%) 0 10.7 80 9.3 0 0 0 0 0 Rhodamine B (%) 0 0 0 0 75.4 24.6 0 0 0 Rhodamine 6G (%) 0 0 0 0 0 44.5 55.5 0 0

[0407] Table 3: Percentage distribution of concentration of each compound in the outputs of the first subunit Ui

[0408] The results of the HPLC analysis at 2500 V reveal: - Fluorescein emerges pure at the 3 outputs Si(2), Si(3) and Si(4), primarily at output Si(3), without signal from rhodamine B and 6G - 75.4% of rhodamine B exits pure at the Si(5) exit - 55.5% of the rhodamine 6G exits pure at the Si(7) exit - An outlet flux Si(6) consisting of a mixture of Rhodamine B (24.6%) and Rhodamine 6G (44.5%)

[0409] Therefore, to optimize the separation, the output Si(6), composed of Rhodamine B (24.6%) and Rhodamine 6G (44.5%) must be treated in the next second subunit in order to finalize the separation of the Rhodamines.

[0410] Furthermore, it is also observed that the concentration of each dye is higher at the outlet of the subunit when the electric field is applied than when it is not, reflecting a concentration phenomenon of the sample during separation and no longer a dilution as previously observed when the electric field is zero. This is also reflected in the visual analysis of the recovered fractions at the outlet.

[0411] Analysis of outgoing flow rates

[0412] Table 4 below shows the distribution of the flow rates of the outgoing flows from the Up subunit

[0413] [Tables4] Output Flow Rate (pL / min) Si(D) 12.0 Si(2) 36.7 Si(3) 37.7 Si(4) 22.6 St(5) 42.0 St(6) 20.0 Si(7) 31.6 Si(8) 33.0 Si(9) 13.6 Total 250.0

[0414] Table 4: Flow rates at the outputs of subunit Ui

[0415] Table 5 reports the flow rate values ​​in mg / min of each molecule at each of the outputs of the subunit Ui, corresponding to the concentration multiplied by the flow rate of the outgoing flux, for a zero electric field.

[0416] [Tables5] Flow rate (mg / min) S4D S42) Si(3) S<(4) Si(5) Si (6) Si(0 Si (8) St (9) Réofescêtne 0 0 0 Û 0.00156 0 0 0 0 Rhodamine B 0 0 û 0 0.00255 i0 0 0 0 Rhodamine6î3 0 û 0 û' 0.00223 0 0 0 0

[0417] Table 5: Flow rate in mg / min of each dye at the outputs of the Ui subunit under zero electric field.

[0418] Table 6 and [Fig. 14] report the flow rate values ​​in mg / min of each molecule at each of the outlets of the Ul subunit, corresponding to the concentration multiplied by the outflow rate, for a zero electric field and for an electric field of 2500 V.

[0419] [Tableauxô] Flow rate (mg / min) S(1) S 42) Si(3) •M4) St(b) St (6) W) St(8) 5,(9) Fluorescein 0 0.000170 0.001266 0.000060 0 0 0 0 0 Rhodamine B 0 0 0 Û 0.001712 0.000560 0 0 0 Rhodamine 6G o 0 0 0 0 0.001023 0.001278 0 0

[0420] Table 6: Flow rate in mg / min of each dye at the outputs of the Ui subunit under an electric field of 2500 V.

[0421] In the present case, the flow rate at the outlet of Si(6) of the first subunit is on the order of 20 pL / min, therefore:

[0422] - rhodamine B present at 0.024 mg / mL is recovered at a rate of 0.00056 mg / min

[0423] - rhodamine 6G present at 0.045 mg / mL is recovered at a rate of 0.0013 mg / min

[0424] Example 3: Second subunit U2 Polishing step by electrophoretic separation

[0425] An electrophoretic separation of the outgoing Si(6) flux comprising a mixture of rhodamine 6G and rhodamine B, as the incoming flux at a rate of 20 pL / min, was then implemented in the second subunit U2 of configuration EAD-008, i.e., of the same configuration as that of subunit Ui, which comprises:

[0426] - n2 = 5 entries

[0427] - m2 = 9 outlets,

[0428] - a height h2 of 100 pm

[0429] - width La2 of 3 cm

[0430] - length Lo2 of 6.4 cm

[0431] The flow rates of the flows injected using the flow meters into the inlets of subunit U2 are, for the mixture to be purified and / or separated, the buffer solutions and the liquid electrode, respectively, 10 / 100 / 20 in pL / min. The inlet solution setup is shown in Table 7 below.

[0432] The applied electric field voltage CE(2) is set at 2500 V, namely the same value as that of the subunit Ub

[0433] [Tables7] Inlets Injected Solution Composition Flow Rate (pL / min) E2(l) Liquid Electrode (Cathode) Water / methanol (70 / 30), 10mM HEPES, 0.2%(w / v) HPMC, 1.5M KCl pH = 7.75 Conductivity 102.6 mS / cm Viscosity (25°C) 2.19 mPa / s 20 E22) Buffer Solution Water 10mM HEPES, 0.2%(w / v) HPMC, 0.1%(w / v) Tween 20 pH = 7.61 Conductivity 393.6 mS / cm 100 Viscosity (25°C) 1.22 mPa / s E2(3) Mixture to be purified and / or separated corresponding to the outgoing Si flux (6) from Up Mixture of 3 dyes: -Fluorescein, -Rhodamine 6G and -Rhodamine B Water 20 E2(4) Buffer solution Water HEPES at 10mM, HPMC at 0.2%(w / v), Tween 20 at 0.1%(w / v) pH = 7.61 Conductivity 393.6 mS / cm Viscosity (25°C) 1.22 mPa / s 100 E2(5) Liquid electrode (Anode) Water / methanol (70 / 30), HEPES at 10mM, HPMC at 0.2%(w / v), KCl 1.5M pH = 7.75 Conductivity 102.6 mS / cm Viscosity (25°C) 2.19 mPa / s 20

[0434] Table 7: Operating conditions in the E2(1) to E2(5) inputs of the U2 subunit

[0435] Thus, the inputs E2(1) and E2(5) were each supplied by a solution The electrolytic solutions for the anode and cathode are identical. The composition is the same as that used for the Up subunit. The liquid electrolytic solution has the following composition: HEPES at 10 mM, HPMC at 0.2% (w / v), methanol at 40%, and KCl 1.5 M. The flow rate was set at 20 pL / min. These electrolytic solutions exhibited a pH of 7.75, a conductivity of 102.6 mS / cm, and a viscosity at 25°C of 2.19 rnPa / s. The electric field is generated by the electrolytic solutions in the electrophoresis chamber, which are the ones containing the carbon electrodes, anode and cathode, respectively.

[0436] The central inlet E2(3) was fed by the flow from the outlet Si(6), which constituted the solution to be purified and / or separated, comprising the mixture of two rhodamines. The flow rate was set at 20 pL / min throughout the separation.

[0437] The two inlets E2(2) and E2(4) were supplied with a buffer solution composed of HEPES at 10 mM, HPMC at 0.2% (nVv), Tween 20 at 0.1% (m / v) in water, at a fixed flow rate of 100 pL / min. The buffer solution had a pH of 7.61, a conductivity of 393.6 pS / cm, and a viscosity at 25°C of 2.19 mPa / s.

[0438] The electric field was fixed in this test at 2500 V. After stabilization of the flows, the products exiting the U2 subunit were collected for 30 minutes in tubes and analyzed by HPLC to determine the percentage of each compound at each exit.

[0439] A control test was carried out with an electric field fixed at 0 V. The system was left running continuously for 30 minutes.

[0440] Example 5 - Result of polishing by electrophoretic separation in the U2 subunit

[0441] At 2500 V, the initially faint beam becomes invisible, preventing visual analysis and photography. Only fraction recovery and HPLC analysis, performed after 30 minutes of sample collection, can determine whether or not the dyes were properly separated. However, the recovered fractions show different coloration in some outputs, particularly in the S2(5), S2(6), and S2(7) fractions.

[0442] For the control, at 0 V, only the outputs of the electrophoresis subunit S2(4) to S2(6) were analyzed.

[0443] The HPLC spectra of the products exiting at the 0V and 2500V outputs are shown in [Fig.15],

[0444] The relative distribution in concentration of each dye in the outputs by HPLC analysis at 2500 V is reported in Table 8 below.

[0445] [Tables8] S2(l) S2(2) S2(3) S2(4) S2(5) S2(6) S2(7) S2(8) S2(9) Rhodamine B (%) 0 0 0 0 80 20 0 0 0 Rhodamine 6G (%) 0 0 0 0 0 44.5 55.5 0 0

[0446] Table 8: Percentage distribution of concentration of each compound in the outputs of the second U2 subunit

[0447] At 2500 V, a separation of the dyes present at the outputs S2(5) to S2(7) is again observed: rhodamine B is pure in the fraction S2(5) which represents a gain of 18% of the total rhodamine to be purified.

[0448] We therefore obtain a purification yield of pure rhodamine B of the order of 88.5% with 2 successive subunits.

[0449] Furthermore, working continuously, it is still possible to recycle the output flux S2 (6) of the 2nd subunit which represents about 11.5% of the initial amount of rhodamine B.

[0450] Analysis of outgoing flow rates

[0451] Table 9 below shows the distribution of the flow rates of the outgoing flows from subunit U2.

[0452] [Tables9] Output Flow (pL / min) S2(l) 12.0 S2(2) 36.7 S2(3) 37.7 S2(4) 22.6 S2(5) 42.0 S2(6) 24.0 S2(7) 33.6 S2(8) 35.0 S2(9) 13.6 Total 250.0

[0453] Table 9: Flow rates at the outputs of subunit U2

[0454] Table 10 reports the flow rate values ​​in mg / min for each molecule at the level of each of the outputs of the second subunit U2, corresponding to the concentration multiplied by the outflow rate, for a zero electric field.

[0455] [TableauxlO] Flow rate (mg / min) S4L &(2) Sz(3) Sz(4) S;(7) Sz(8) SpS) Ffeorescétne 0 0 0 0 0 0 n 0^ 0 Rhodamine B 0 0 0 0 0..000576 Ô 0 0 0 Rhodamine 6G 0 0 0 0 0.001023 0 0 0 0

[0456] Table 10: Flow rate in mg / min of each dye at the outlets of the second subunit U2 under zero electric field.

[0457] Table 11 and [Fig. 16] report the flow rate values ​​in mg / min of each molecule at each of the outputs of the subunit Ui, corresponding to the concentration multiplied by the flow rate of the outgoing flux, for a zero electric field and for an electric field of 2500 V.

[0458] [Tables 11] Flow rate: (mg / min) 82(1) SF7Ï Ss(3) 82(4) S2(5) &(§) 82(7) Ss / S) Sz(9) Fluorescein g 0 Ô 0 0 0 0 0 0 Rhodamine B- U 0 0 <3 0.000448 1-003:12 0 0 o Rhodamine6G $ 0 ô 0 0 0.0-00455 0.000568 0 0

[0459] Table 11: Flow rate in mg / min of each dye at the outlets of the second subunit U2 under an electric field of 2500 V.

[0460] Example 6: Separation Summary

[0461] Table 12 below reports the separation yield of each compound in the mixture and the percentage of compounds still in mixture form.

[0462] [Tables 12] Initial composition mg / mL Separation yield Percentage remaining to be recycled Fluorescein 0.1559 100% 0 Rhodamine B 0.2502 88% 12% Rhodamine 6G 0.2291 80.5% 19.5%

[0463] Table 12: Separation efficiency

[0464] Thus, the cell comprising two subunits of EAD 008 configuration made it possible to purify the mixture composed of fluorescein, rhodamine B, and rhodamine 6G with excellent separation yields of 100% for fluorescein, 88% for rhodamine B, and 80% for rhodamine 6G. These three compounds are obtained diluted in an aqueous medium and can be concentrated to obtain their pure form.

[0465] In conclusion, the use of a cell comprising a chamber of two successive cooperating subunits allows for the optimization of separation and purification. In a mixture of three dyes, it was possible to eliminate a greater proportion of the unwanted compounds (fluorescein and rhodamine 6G) in favor of the compound of interest, in this case rhodamine B. Furthermore, the free-flow electrophoresis cell allows for the adaptation and optimization of operating conditions and configurations to obtain greater purification and / or separation in continuous flow.

[0466] The adaptability of the electrophoresis cell allows a configuration introducing a third subunit to increase the purification and / or separation yield of rhodamines B and 6G by separating and purifying the remaining percentage to be recycled contained in the S2(6) output of the U2 subunit.

Claims

Demands

1. Free-flow electrophoresis cell for purifying and / or separating an initial solution containing a product to be purified and / or separated, comprising - at least one plate X between two adjacent plates Y: X being an electrophoresis plate (1) of inert material, Y being a sealed plate (2) of sapphire or alumina A12O3 with 99% a-Al2O3, an electrophoresis chamber comprising s subunits Ui, where 5 represents an integer from 2 to 5 and i is an integer ranging from 1 to s, in which each subunit U; comprises • a hollowed-out section in a closed plate X sandwiched between two plates Y, • in the shape of a rectangular parallelepiped with 4 lateral faces (a, b, Ci, d) and 2 upper and lower faces (ei5 fj), • said hollowed-out part being of length Lo; and of width La;, • of height h; corresponding to the thickness of the X-ray electrophoresis plate, from 25 µm to 20 mm, • the lateral faces (a;, b;) being parallel to each other, the face (a;) being delimited by two edges (Al;, A2;) of dimension La; and the face (b;) being delimited by two edges (Bli, B2i) of dimension La;, • the lateral faces (c;, d;) being parallel to each other, the face (Cj) being delimited by two edges (Cl;, C2;) of dimension Lo; and the face (d;) being delimited by two edges (DI;, D2i) of dimension Lo;, • n; successive entries E;(1), E;(2) to Ei(n;-1), Ei(n;), n; being an integer from 4 to 9, distributed on the face (a;

2. ) between Ah and A2; and aligned along a direction parallel to Ah and A2; • m successive outputs from S;(l), S;(2) to S^rUi-l), Si(mi), ni; being an integer from 4 to 12, distributed on the face (b;) between B h and B2; and aligned along a direction parallel to B h and B2;, so that S;(l) faces Ei(l) and Si(nii) faces Ei(n;) along a direction parallel to Ch and Dh, - a cooling circuit, characterized in that the electrophoresis chamber comprises: - (s-1) intermediate channels Vk, k being an integer ranging from 1 to (s-1), each channel Vk connecting the two successive subunits Uk and Uk+i via one of the outputs SVk(Uk) chosen from Sk(2) to Sk(mk-1) of the subunit Uk to one of the inputs EVk(Uk+i) of the subunit Uk+ chosen from Ek+1(2) to Ek+1(nk+1-l). Cell according to claim 1, comprising: - supply channels configured to connect the 5 inputs E;(l), i varying from 1 to s, from each subunit of the electrophoresis chamber to at least one external micro / milli fluidic circuit supplying at least one liquid cathode, - supply channels configured to connect the E(n) inputs of each subunit of the electrophoresis chamber to at least one external micro / millifluidic supply circuit for at least one liquid anode, - a supply channel configured to connect at least one of the inputs Ei(2) to Ei(nrl) of the subunit Ui to an external micro / milli fluidic circuit supplying an initial solution containing a product to be purified and / or separated, - Supply channels configured to connect at least one input of each electrophoresis chamber subunit to at least one external micro / milli circuit fluidic supply of at least one buffer solution, - at least one recovery channel configured to connect one of the outputs of the Us subunit chosen from Ss(2) to Ss (ms-l) of the electrophoresis chamber to an external micro / milli fluidic recovery circuit.

3. A cell according to either claim 1 or 2, said cell being configured to: • in each subunit Ui,, in the presence of an electric field generated CE(i) between a liquid cathode CAL(i) and a liquid anode ANL(i) parallel to Al; and perpendicular to Cl;, • and in operation, a) in the subunit Ui: • circulate the liquid cathode CAL(l) from the inlet Ei(l) to the outlet Si(l), • circulate the liquid anode ANL(l) from the inlet Ei(ni) to the outlet Si(mi), • circulate the initial solution containing the product to be separated and / or purified and at least one buffer solution in the Ui subunit, between the liquid cathode CAL(l) and the liquid anode ANL(l), from the inlets Ei(2) to Ei(nrl) to the outlets Si(2) to Si(mi-l), to obtain the purified and / or separated product from the Ub subunit b) in each intermediate channel Vk, k varying from 1 to (s-1), • recover at the SVk(Uk) outlet the purified and / or separated product from the Uk subunit; • to circulate in channel Vk the aforementioned product from the output SVk(Uk) of the subunit Uk, from SVk(Uk) to the input EVk(Uk+i) of the subunit Uk+i, c) in the subunit Uk+i: • circulate the liquid cathode CAL(k+l) from the inlet Ek+[ (1) to the outlet Sk+i(l), • circulate the liquid anode ANL(k+l) from the inlet Ek+i(nk+i ) to the outlet Sk+i(mk+i), • circulate said purified and / or separated product from the outlet SVk(Uk) of the subunit Uk and at least one buffer solution in the subunit Uk+i, between the liquid cathode CAL(k+l) and the liquid anode ANL(k+l), from the inlets Ek+i(2) to Ek+1(nk+1-l) to the outlets Sk+i(2) to Sk+i(mk+1-l), d) recover at one of the outlets Ss(2) to Ss(ms-1) the purified and / or separated product.

4. Cell according to any one of claims 1 to 3, further comprising: at least one complementary intermediate channel W said complementary intermediate channel W connects two successive subunits Uq and Uq+i, q being an integer from 1 to (s-1), said channel W extending: - from one of the outputs of unit Uq chosen from Sq(2) to Sq(mq- 1), different from the output linked to channel Vq, - to the input linked to channel Vq of subunit Uq+i, said input of channel Vq being connected to at most (mq-3) output(s) chosen from Sq(2) to Sq(mq-1).

5. Cell according to any one of claims 1 to 4, comprising a plate X between two adjacent plates Y, said plate X comprising the 5 hollowed-out parts of the U subunits; of said electrophoresis chamber, the height of the hollowed-out part of each subunit being of identical value, or comprising at least two plates X, each plate X being between two adjacent plates Y, each plate X comprising at least one hollowed-out part of a U subunit; of said electrophoresis chamber.

6. Cell according to any one of claims 1 to 5, comprising at least one cooling plate Z comprising a heat transfer system, said plate Z being adjacent to one of the two plates Y adjacent to said plate X, and / or comprising at least two plates Z comprising a heat transfer system, said cell comprising the following plate sequence ZYXYZ, in particular the sequence YZYXYZY, in particular wherein said heat transfer system is a network of pipes configured to permit the circulation of one or more heat transfer fluids parallel to the edges Q or perpendicular to the edges C of the subunits Ui, in particular said network of pipes comprising inlets and outlets of the heat transfer system, engraved in plate Z.

7. Cell according to any one of claims 1 to 6, wherein the subunits U; of the electrophoresis chamber comprise means for channeling the flows opening onto the inlets E;(l) to Ei(n;) and / or onto the outlets S;(l) to Si(mi) of the subunits U; of the electrophoresis chamber, said channeling means preferably being etched into the electrophoresis plate X, said means in particular being triangular in shape, situated between two adjacent inlets or two adjacent outlets of a subunit, and / or wherein the width La; of each of the s subunits U; is from 1.0 to 8.0 cm, and / or the length Lo; of each of the s subunits U; the electrophoresis chamber is 2.0 to 20.0 cm, and / or in which the electrophoresis plate X is made of material selected from polytetrafluoroethylene (PTFE), perfloroalkoxy (PFA) and fluoroethylene propylene (FEP), in particular Teflon™, Teflon™-PFA and Teflon™-FEP plates.

8. A cell according to any one of claims 1 to 7, wherein 5 is equal to 2, the electrophoresis chamber comprising two subunits Ui and U2 - the subunit Ui comprising: • a portion hollowed out of a plate X • in the form of a rectangular parallelepiped with 4 lateral faces (ab bh cb dj) and 2 upper and lower faces (eb fi), • said hollow portion having length Loi and width Lab • of height hi corresponding to the thickness of the electrophoresis plate X, from 25 µm to 20 mm, • the lateral faces (ab bj) being parallel to each other, the face (aj) being delimited by two edges (Alb A2i) of dimension Lai and the face (bj) being delimited by two edges (B1bB2i) of dimension Lab • the lateral faces (cb dj) being parallel to each other, the face (ci) being delimited by two edges (Clb C2i) of dimension Loi and the face (dj) being delimited by two edges (Dlb D2i) of dimension Lob • ni successive entries E^l), Ei(2) to Ei(nrl), Ei (nj, ni being an integer from 4 to 9, distributed on the face (aj between Ali and A2i and aligned along a direction parallel to Ali and A2b • mi successive exits from Si(1), Si(2) to Si(mi-1), Si(mi), where mi is an integer from 4 to 12, distributed on the face (bi) between B1 and B2i and aligned along a direction parallel to B1 and B2b so that Si(l) faces Ei(l) and Si(mi) faces Ei(ni) along a direction parallel to C1 and DIb The U2 subunit comprises • a hollowed-out portion in a plate X, possibly in the aforementioned plate X of the aforementioned subunit Ub • in the shape of a rectangular parallelepiped with 4 lateral faces (a2, b2, c2, d2) and 2 upper and lower faces (e2, f2), • said hollowed-out part being of length Lo2 and width La2, • of height h2 corresponding to the thickness of the X-ray electrophoresis plate, from 25 µm to 20 mm, • the lateral faces (a2, b2) being parallel to each other, the face (a2) being delimited by two edges (Al2, A22) of dimension La2 and the face (b2) being delimited by two edges (Bl2, B22) of dimension La2, • the lateral faces (c2, d2) being parallel to each other, the face (c2) being delimited by two edges (Cl2, C22) of dimension Lo2

9. and the face (d2) being delimited by two edges (Dl2, D22) of dimension Lo2, • n2 successive entries E2(1), E2(2) to E2(n2-1), E(n2), n2 being an integer from 4 to 9, distributed on the face (a2) between Al2 and A22 and aligned along a direction parallel to Al2 and A22, • m2 successive exits from S2(l), S2(2) to S2(m2-1), S2(m2), m2 being an integer from 4 to 12, distributed on the face (b2) between Bl2 and B22 and aligned along a direction parallel to B12 and B22, so that S2(l) faces E2(l) and S2(m2) faces E2(n2) along a direction parallel to Cl2 and Dl2, characterized in that it includes - an intermediate channel Vb connecting the two subunits Ui and U2 by one of the outputs SVi(Ui) chosen from Si(2) to Si(mrl) of the subunit Ui to one of the inputs EVi(U2) of the subunit U2 chosen from E2(2) to E2(n2-1). Cell according to claim 8, comprising: - a supply channel configured to connect the Ei(l) inlet of the electrophoresis chamber subunit Ui to an external micro / milli fluidic supply circuit for a first liquid cathode CAL(l), - a supply channel configured to connect the Ei(nO) inlet of the Ui subunit of the electrophoresis chamber to an external micro / milli fluidic supply circuit for a first liquid anode ANL(l), - a supply channel configured to connect at least one of the inputs Ei(2) to Ei(nrl) of the subunit Ui to an external micro / milli fluidic circuit for supplying an initial solution containing a product to be purified and / or separated, - at least one supply channel configured to connect to at least one remaining input of the Ui unit at least one external micro / millifluidic supply circuit for at least one buffer solution, - a supply channel configured to connect the E2(l) inlet of the U2 electrophoresis chamber subunit to an external micro / millifluidic supply circuit for a second liquid cathode CAL(2), - a supply channel configured to connect the E2(n2) inlet of the U2 electrophoresis chamber subunit to an external micro / millifluidic supply circuit for a second liquid anode ANL(2), - at least one supply channel configured to connect at least one remaining inlet of the U2 unit to at least one external micro / millifluidic supply circuit for at least one buffer solution, - at least one recovery channel configured to connect one of the outputs of the U2 subunit selected from S2(2) to S2(m2-l) of the electrophoresis chamber to an external micro / millifluidic supply circuit recovery, said cell being configured for, in the presence of: • of an electric field generated CE(1) in the subunit Ui between the liquid cathode CAL(l) and the liquid anode ANL(l) parallel to Ali and perpendicular to Cli, • of an electric field generated CE(2) in the U2 subunit between the liquid cathode CAL(2) and the liquid anode ANL(2) parallel to Al2 and perpendicular to Cl2, • and in operation, a) in the subunit Ui: • circulate the liquid cathode CAL(l) from the inlet Ei(l) to the outlet Si(l), • circulate the liquid anode ANL(l) from the inlet Ei(ni) to the outlet Si(mi), • circulate the initial solution containing the product to be separated and / or purified and at least one buffer solution in the Ub subunit between the liquid cathode CAL(l) and the liquid anode ANL(l), from the inputs Ei(2) to Ei(nrl) to the outputs Si(2) to Si(mi-l), b) in the intermediate channel Vb • recover at the output SVi(Ui) the purified and / or separated product contained in said initial solution; • circulate the purified and / or separated product from the SVi(Ui) outlet of the Ub subunit of SV(Ui) through channel Vi to the EVi(U2) inlet of subunit U2, c) in subunit U2: • circulate the liquid cathode CAL(2) from the E2(l) inlet to the S2(l) outlet, • circulate the liquid anode ANL(2) from the E2(n2) inlet to the S2(m2) outlet, • circulate the purified and / or separated product from the SVi(Ui) outlet of subunit Ui and at least one buffer solution in subunit U2, between the liquid cathode CAL(2) and the liquid anode ANL(2), from inlets E2(2) to E2(n2-l) to outlets S2(2) to S2(m2-1), d) recover at one of the S2(2) outlets S2(m2-1) the purified and / or separated product.

10. Cell according to any one of claims 8 or 9, wherein the number of outputs m, of unit Ui is greater than or equal to 5, further comprising an intermediate channel Wi linking subunit Ui to subunit U2, extending: • from one of the outputs SWi(Ui) of unit Ui chosen from Si (2) to Si(mi-1), different from the output SVi(Ui) of channel Vb • to the input EVi(U2) of subunit U2.

11. A cell according to any one of claims 8 to 10, wherein the number of outputs mi and m2 of subunits Ui and U2 is greater than the number of inputs ni and n2 of subunits Ui and U2, respectively; and / or wherein subunits Ui and U2 have width and length dimensions such that the width La2 of subunit U2 is equal to the width Lai of subunit Ui and the length Lo2 of subunit U2 is equal to the length Loi of subunit Ui, or in which the width La2 of subunit U2 is greater than or equal to the width Lai of subunit Ui and the length Lo2 of subunit U2 is less than or equal to the length Loi of subunit Ub or in which the width La2 of the subunit U2 is less than or equal to the width La1 of the subunit Ui and the length Lo2 of the subunit U2 is greater than or equal to the length Lo2 of the subunit Ui, or in which the width La2 of the subunit U2 is less than or equal to the width Lai of the subunit Ui and the length Lo2 of the subunit U2 is less than or equal to the length Loi of the subunit Ub or in which the width La2 of the subunit U2 is greater than or equal to the width La! of the subunit Ui and the length Lo2 of the subunit U2 is greater than or equal to the length Loi of the subunit Ui.

12. Cell according to any one of claims 1 to 7, wherein s is equal to 3, the electrophoresis chamber comprising three subunits Ub, U2 and U3, The Ui subunit includes • a hollowed-out section in a plate X • in the shape of a rectangular parallelepiped with 4 lateral faces (ab bb cb di) and 2 upper and lower faces (eb fi), • said hollowed-out part being of length Loi and width Lab • of height hi corresponding to the thickness of the X-ray electrophoresis plate, from 25 µm to 20 mm, • the lateral faces (ai, bi) being parallel to each other, the face (ai) being bounded by two edges (Alb A2i) of dimension Lai and the face (bi) being bounded by two edges (Blb B2i) of dimension Lab • the lateral faces (cb dj) being parallel to each other, face (ci) being delimited by two edges (Clb C2i) of dimension Loi and face (di) being delimited by two edges (Dlb D2i) of dimension Lob • ni successive entries E^l), Ei(2) to Ei(nrl), Ei (nj, ni being an integer from 4 to 9, distributed on the face (aj between Ali and A2i and aligned along a direction parallel to Ali and A2b • mi successive exits from Si(1), Si(2) to Si(mrl), Si(mi), where mi is an integer from 4 to 12, distributed on the face (bi) between B1 and B2i and aligned along a direction parallel to B1 and B2b so that Si(l) faces Ei(l) and Si(mi) faces Ei(ni) along a direction parallel to C1 and DIb The U2 subunit comprises • a hollowed-out section in a plate X • in the shape of a rectangular parallelepiped with 4 lateral faces (a2, b2, c2, d2) and 2 upper and lower faces (e2, f2), • said hollowed-out part being of length Lo2 and width La2, • of height h2 corresponding to the thickness of the X-ray electrophoresis plate, from 25 µm to 20 mm, • the lateral faces (a2, b2) being parallel to each other, the face (a2) being delimited by two edges (Al2, A22) of dimension La2 and the face (b2) being delimited by two edges (Bl2, B22) of dimension La2, • the lateral faces (c2, d2) being parallel to each other, the face (c2) being delimited by two edges (Cl2, C22) of dimension Lo2 and the face (d2) being delimited by two edges (Dl2, D22) of dimension Lo2, • n2 successive entries E2(1), E2(2) to E2(n2-1), E(n2), n2 being an integer from 4 to 9, distributed over the face (a2) between Al2 and A22 and aligned along a direction parallel to Al2 and A22, • m2 successive exits from S2(l), S2(2) to S2(m2-1), S2(m2), m2 being an integer from 4 to 12, distributed on the face (b2) between Bl2 and B22 and aligned along a direction parallel to B12 and B22, so that S2(l) faces E2(l) and S2(m2) faces E2(n2) along a direction parallel to Cl2 and Dl2, The U3 subunit comprises • a hollowed-out section in a plate X • in the shape of a rectangular parallelepiped with 4 lateral faces (a3, b3, c3, d3) and 2 upper and lower faces (e3, f3), • said hollowed-out part being of length Lo3 and width La3, • of height h3 corresponding to the thickness of the X-ray electrophoresis plate, from 25 µm to 20 mm, • the lateral faces (a3, b3) being parallel to each other, the face (a3) ​​being delimited by two edges (Al3, A23) of dimension La3 and the face (b3) being delimited by two edges (Bl3, B23) of dimension La3, • the lateral faces (c3, d3) being parallel to each other, the face (c3) being delimited by two edges (Cl3, C23) of dimension Lo3 and the face (d3) being delimited by two edges (Dl3, D23) of dimension Lo3, • n3 successive entries E3(1), E3(2) to E3(n3-1), E(n3), n3 being an integer from 4 to 9, distributed on the face (a3) ​​between Al3 and A23 and aligned along a direction parallel to Al3 and A23, • m3 successive outputs from S3(1), S3(2) to S3(m3-1), S3(m3), m3 being an integer from 4 to 12, distributed on the face (b3) between B13 and B23 and aligned along a direction parallel to B13 and B23, so that S3(l) faces E3(l) and S3(m3) faces E3(n3) in a direction parallel to Cl3 and Dl3, characterized in that it comprises - an intermediate channel Vb connecting the two subunits Ui and u2 by one of the outputs SVi(Ui) chosen from Si(2) to Si(mrl) of the subunit Ui to one of the inputs EVi(U2) of the subunit U2 chosen from E2(2) to E2(n2-1), and - an intermediate channel V2, connecting the two subunits U2 and u3 by one of the outputs SV2(U2) chosen from S2(2) to S2(m2-1) of the subunit U2 to one of the inputs EV2(U3) of the subunit U3 chosen from E3(2) to E3(n3-1).

13. Free-flow electrophoresis cell device comprising: - p plates X between two adjacent plates Y: • p being an integer from 1 to 50, in particular from 1 to 20, • X being an electrophoresis plate (1) of inert material, • Y being a sealed plate (2) of sapphire or alumina Al12O3 with 99% a-Al2O3, - f electrophoresis cells according to any one of claims 1 to 12, f is an integer from 1 to 500, - clamping means for all the plates enabling the sealing of said device.

14. Device according to claim 13, comprising at least two electrophoresis cells such that in at least one of their subunit Ut, t being an integer from 1 to (s-1), at least two of their respective outputs open into the same input of the same successive subunit Ut+i through their respective channel Vt.

15. Device according to any one of claims 13 or 14, wherein each plate X named Xj, j being an integer ranging from 1 to p, comprises q, electrophoresis cells according to any one of claims 1 to 12, q, being an integer from 1 to 10.

16. A device according to claim 15, wherein - p is an integer from 1 to 50, in particular from 1 to 10, - when j varies from 1 to p, q is an integer from 2 to 50, in particular from 2 to 10, comprising p electrophoresis plates Xj, each having from 2 to 50 electrophoresis chambers, in particular from 2 to 10 electrophoresis chambers, each electrophoresis chamber having 5 subunits, 5 being an integer from 2 to 5, wherein for each electrophoresis chamber, the hollowed-out portions of the 5 subunits are located in the same plate, or wherein p is equal to 1 and qi is an integer from 2 to 50, in particular from 2 to 10, comprising a single electrophoresis plate Xi having from 2 to 50 electrophoresis chambers, in particular from 2 to 10 chambers of electrophoresis, each electrophoresis chamber comprising 5 subunits, 5 representing an integer from 2 to 5, and / or in which in each plate Xj, the q,electrophoresis chambers are aligned so that the Uides q subunits, electrophoresis chambers are adjacent to each other by the (cO or (d;) faces.

17. A device according to any one of claims 13 to 16, comprising a vertical succession of plates X, Y, Z whose surfaces are stacked in the sequence YZY(XYZY)p, in which X represents an electrophoresis plate made of an inert material, Y represents a sealed plate made of sapphire or alumina Al2O3 with 99% a-Al2O3, Z represents a cooling plate comprising a heat transfer system, p being an integer from 1 to 50, in particular from 1 to 20, representing both the number of stages of said device and the number of plates X, each stage being defined: - by the following sequence of plates YZYXYZY, in which: - plate X is located between two plates Y, - each of the two plates Z being respectively adjacent to a plate Y, - and each of the two Y plates located at the ends of the YZYXYZY sequence, respectively covers a Z plate so that each Z plate is located between two Y plates.

18. A device according to any one of claims 13 to 17, wherein each electrophoresis chamber is configured to contain at least one selective permeability membrane, in particular size-selective, positioned so as to be traversed by the solution containing the product to be separated or purified during operation of the device, preferably said at least one selective permeability membrane is placed in the Us subunit of each electrophoresis chamber; and / or wherein the upper faces e; and / or the lower faces fi of the U subunits;of each of the electrophoresis chambers include at least one protrusion configured not to disturb, during the operation of the device, the circulation in the chamber of the product to be purified and / or separated, and configured to improve heat transfer and to maintain each of said electrophoresis chambers at a selected temperature, in particular wherein said at least one protrusion is made of thermally conductive material, preferably of sapphire or 99% alumina a-Al2O3; and / or said device includes a supply channel network configured to connect at least one of the inlets Ei(2) to Ei(nrl) of the subunit Ui of each electrophoresis chamber to an external micro / milli fluidic circuit for supplying an initial solution containing a product to be purified and / or separated.

19. A method for purifying and / or separating, by free-flow electrophoresis, a product to be purified and / or separated contained in an initial solution by implementing an electrophoresis cell device according to any one of claims 13 to 18, and comprising the following steps: a) a purification and / or separation step of said product in the subunits Ui of each electrophoresis chamber, under an electric field CE(1), comprising: in the subunit Ui of each electrophoresis chamber: • the circulation of a liquid cathode CAL(l) from the inlet Ei (1) to the outlet Si(l), • the circulation of a liquid anode ANL(l) from the inlet Ei(ni ) to the outlet Si(mi), • the circulation of the initial solution containing the product to be separated and / or purified and at least one buffer solution in the Ub subunit between the liquid cathode CAL(l) and the liquid anode ANL(l), from the inlets Ei(2) to Ei(nrl) to the outlets Si(2) to Si(mrl), to obtain the purified and / or separated product from the Ui subunit, b) (s-1) polishing step(s) k, k ranging from 1 to (s-1), each step k comprising each: i) in the intermediate channel Vk of each electrophoresis chamber, • the recovery at the SVk(Uk) outlet corresponding to the intermediate channel Vk of said purified and / or separated product from the Uk subunit; • the circulation, in the absence of an electric field, of the aforementioned purified and / or separated product, from the output SVk(Uk) of the subunit Uk corresponding to the intermediate channel Vk to the input EVk(Uk+i) of the subunit Uk+i corresponding to the intermediate channel Vk, ii) in the Uk+i subunit of each electrophoresis chamber, in the presence of an electric field CE(k+l): • the circulation of a liquid cathode CAL(k+l) from the inlet Ek+i(l) to the outlet Sk+i(l), • the circulation of a liquid anode ANL(k+l) from the inlet Ek +i(nk+i) to the outlet Sk+i(mk+i), • the circulation of said purified and / or separated product from the outlet of the Uk subunit and at least one buffer solution in the Uk+i subunit, between the liquid cathode CAL(k+l) and the liquid anode ANL(k+l), from the inlets Ek+[(2) to Ek+1(nk+1-l) to the outlets Sk+i(2) to Sk+i(mk+1-l), to obtain the product (k+1) times purified and / or separated from the subunit Uk+i; c) a recovery step comprising: - the recovery at one of the outputs Ss(2) to Ss(ms-1) of the unit Us of each electrophoresis chamber of the product(s) times purified and / or separated.

20. Use of an electrophoresis cell according to any one of claims 1 to 12 or of a device according to any one of claims 13 to 19, in the implementation of a process of purification and / or separation by free flow electrophoresis, of a product to be purified and / or separated contained in an initial solution, comprising a first step of purification and / or separation in the subunit(s) Ui followed by (s-1) successive polishing step(s) in the subunits U2 to Us of the electrophoresis chamber(s).

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