CN1553192A

CN1553192A - Magnetic separator for immunological analytical separation

Info

Publication number: CN1553192A
Application number: CNA031383491A
Authority: CN
Inventors: 林金明; 赵利霞; 屈锋
Original assignee: Research Center for Eco Environmental Sciences of CAS
Current assignee: Research Center for Eco Environmental Sciences of CAS
Priority date: 2003-05-27
Filing date: 2003-05-27
Publication date: 2004-12-08
Anticipated expiration: 2023-05-27
Also published as: CN1231759C

Abstract

A magnetic separator consists of top yoke iron plate and bottom yoke iron plate which are placed in parallel, magnetic plate which is located in between the two and three being in the same size, a piece of plate frame which is set on each side face of bottom yoke iron plate with barb bending towards inside. The space distance between two frames is 7-10.50 cm and space distance between top yoke iron plate and barb bottom end is 1.0-2.0 cm. The magnetic plate is made by permanent ferrite, soft magnetic ferrite, electromagnet or some other materials.

Description

A kind of immunoassay separates uses magnetic separator

Technical field

The invention belongs to the separation vessel in the immunoassay isolation technics, particularly a kind of be applicable in the orifice-plate type magnetic immuno Analytical Separation magnetic separator.

Background technology

Since nineteen fifty-nine, Yallow and Berson have invented after the immunoassay, and immunoassay becomes main analysis tool day by day in clinical chemistry and environmental monitoring field.Immunoassay has homogeneous immunoassay and heterogeneous immunoassay.Fast, the convenient and easily robotization of homogeneous immunoassay speed, but in actual applications and successful unlike the latter.Heterogeneous immunoassay comprises one combination and free part separation steps in addition.Therefore, tripping device plays key effect in heterogeneous immunoassay.The separation method of Shi Yonging is electrophoresis and chromatography the earliest, and they are to separate according to bulk of molecule and charge differences, only are applicable to smaller molecule.The shortcoming of these class methods is non-specific adsorption height, and is simultaneously also very time-consuming.Afterwards, 70 and the glucosan that grown up the eighties apply absorbent charcoal method and precipitation classification.Although this method is practical, non-specific adsorption is still very high, needs refrigerated centrifuge equipment simultaneously.In the present heterogeneous immunoassay, generally adopt two anti-or anti-immobilizations, the solid phase carrier of use comprises magnetic particle, tunica fibrosa, polystyrene tube, microwell plate etc.In these several solid phase carriers, it is big that magnetic material has a surface area, and it is fast to catch analytic target, can effectively separate and advantage such as washing, is subjected to immunologist's favor most.Therefore, thing followed immunomagnetic isolation (IMS) technology is also assented and is given birth to.

In immunoassay, employed any separation method and tripping device all should be simply, fast, expend lowly, and manyly as far as possible must be applicable to more analytic target.At present, in the immunoassay, the separation method of public use and device have following several:

(1) board-like separation method and device: antibody or antigen by physisorption or covalently bound mode coated elisa plate, are added antigen or antibody then, after the immune response, remove supernatant, washing can be finished separation.This isolation technics is used in stronger ELISA Plate of adsorbability or tissue culturing plate, but reaction velocity is slow, and package amount is difficult for evenly.Staimer, N. wait the people that antigen is covalently bound on the ELISA Plate of 96-hole by N-hydroxyl succinimide ester method, use enzyme immunoassay method that the phenyl-β in the human urine-D-thioglucuronide is analyzed, Darwish, I.A. wait the people with antibody by the physisorption coated elisa plate, use a step immunoassay to the Cd in the serum ²⁺Study.(Norbert?Staimer，Shirley?J.Gee，Bruce?D.Hammock，Development?of?asensitive?enzyme?immunoassy?for?the?detection?of?phenyl-β-D-thioglucuronide?in?humanurine，Fresenius.J.Anal.Chem.，2001，369，273-279；Ibrahim?A.Darwish?and?Diane?A.Blake，Development?and?Validation?of?OneOStep?Immunoassay?for?Determination?ofCadminm?in?Human?Serum，Anal.chem.，2002，74，52-58.)。

(2) tubular type magnetic separation technique and device: at present, the Beijing Bei Ai Kang Biotechnology Co., Ltd is commercialization, public use the tubular type magnetic separator.This tubular type magnetic separator is at the bottom of each polystyrene round tube, is placed with a corresponding circular small magnet; After in immunological assay reagents adds pipe, test tube rack is placed on the magnetic separator, magnetic bead is deposited to the bottom in hole under the action of a magnetic field, will manage then with magnetic separator together back-off to clean filter paper, lay equal stress on kowtow bottom, remove supernatant.This design is suitable for manually-operated and separates, washs.(Beijing Bei Ai Kang Biotechnology Co., Ltd, HCG SEROZYME, CODE:10331504).

The most frequently used tripping device be magnetic two kinds in tripping device and the separated device of board-like bag.The magnetic resolution device that immunomagnetic isolation combines with microwell plate also progressively grows up and is subjected to increasing concern.The key of this technology is the design of magnetic separator.The magnetic field of the magnetic separator of general design or be scalariform, or balanced.Dissimilar separation vessels is used in the ELISA Plate of different hole counts, and then understands the efficient of impact analysis.

Summary of the invention

The objective of the invention is to be to provide a kind of immunoassay to separate and use magnetic separator; This magnetic separator uses conjuncted magnet as magnetic sheet, all produces identical magnetic induction density in each space, and is applied widely.Applicable to 6-, 12-, 24-, 32-, 40-, 48-, 96-, 192-, the different hole counts of all equidimensions such as 384-hole comprise transparent and opaque flat ELISA Plate or tissue culturing plate; Be applicable to various analyzing detecting method; Magnetic material can use all its magnetic induction density such as permanent magnet or electromagnet all greater than the material of 2000Hz; This magnetic

Separation vessel is made and is used all very simply, convenient, and velocity of separation is fast, efficient is high, will have broad application prospects in clinical chemistry, environmental pollution and to large-scale epidemic disease context of detection.

Technical scheme of the present invention is as follows:

Immunoassay provided by the invention separates uses magnetic separator, comprise that parallel placement, big plane stack and clamp the be magnetic last yoke plate 1 and the lower yoke plate 3 of plate 2 therebetween, magnetic sheet 2, last yoke plate 1 and lower yoke plate 3 consistent size, each fixes the board-like framework 4 of a strip on the two sides of described lower yoke plate 3, the barb that has turning-in on the upper surface of the board-like framework 4 of described strip, distance between the board-like framework 4 of described two strips is 7-10.50cm, leaves the 1.0-2.0cm spacing between the lower surface of last yoke plate 1 and described barb;

Described magnetic sheet 2 is the magnet plate of being made by ndfeb magnet, permanent ferrite, electromagnet, soft magnetic ferrite, aluminium nickel cobalt or SmCo magnetic material, or is a monoblock/several magnet plates that magnet plate is spliced.

During use, when all immune responses finish, in the time of need separating, ELISA Plate is inserted into the centre of magnetic separator from a side, magnetic particle all is adsorbed onto in the hole of ELISA Plate under the action of a magnetic field.After 1 minute,, supernatant is poured out, tipped upside down on then on the clean filter paper, gently residual liquid is deducted the related separation vessel back-off of ELISA Plate.Extract out from a side, can finish separation.

The present invention has following advantage compared with prior art:

The magnetic separator that is applicable in the orifice-plate type magnetic immuno Analytical Separation provided by the invention, can with ELISA Plate or tissue culturing plate's coupling of various different hole counts.Compare with known magnetic separator, simple, convenient, separation efficiency height, speed be fast, use widerly.Especially can be used for 384-hole or more ELISA Plate or tissue culturing plate's detection of porous plate number, this magnetic separator can carry out large batch of sample detection, has improved analysis efficiency greatly.To have broad application prospects in clinical chemistry, environmental pollution and large-scale epidemic disease context of detection.

Description of drawings

Fig. 1 is a structural representation of the present invention;

Fig. 2 is the synoptic diagram of magnetic sheet of the present invention;

Fig. 3 is the synoptic diagram of framework of the present invention;

Fig. 4 is the structural representation of upper and lower yoke plate of the present invention;

Wherein: go up yoke plate 1 lower yoke plate 2 magnetic sheets 3 frameworks 4

Hole 5

Embodiment

Embodiment with the magnetic separator of the present invention that is suitable for 96-hole ELISA Plate or tissue culturing plate is an example:

The making of the magnetic separator of present embodiment:, have facilitation for the splicing of magnet according to the magnetic conduction mechanism of yoke.We are spliced into a magnetic sheet 2 (also can directly use a magnetic sheet that is of a size of 105mm * 148mm * 8.5mm105mm * 148mm * 8.5mm) with the ndfeb magnet plate of two 105mm * 74mm * 8.5mm, be placed on the lower yoke plate 3 of a 105mm * 148mm * 8.5mm, put one above the magnetic sheet 2 again and go up yoke plate 1, and then will go up yoke plate 1, lower yoke plate 2 and yoke plate 3 be fixed together (upper and lower, left and right must alignment, consistent size); Simultaneously on two long side surfaces of lower yoke plate 3, punch 5 respectively; Article two, the board-like framework 4 of strip of aluminium alloy making (length is 148mm, highly is 28mm) corresponding site has corresponding hole, in order to the board-like framework 4 of two strips is separately fixed at the dual-side of lower yoke plate 3, promptly finishes the making of magnetic separator of the present invention; The top width that the upper surface of the board-like framework 4 of described strip is provided with turning-in is about the barb of 15mm, and this barb lower surface is 1.0-2.0cm apart from the distance between upper surface of last yoke plate 1, and the distance between the board-like framework 4 of two strips is 7-10.50cm.

Can be according to the flat ELISA Plate that is suitable for or the model and the size of tissue culturing plate, especially the design of width and gauge is designed to different sizes such as 7cm, 10.50cm, 8cm or 9cm with the distance between the board-like framework 4 of two strips, the spacing between the lower surface of last yoke plate 1 and described barb is stayed be designed to 1.0cm, 2.0cm, 1.20cm or 1.50cm equidimension;

During use, immunological assay reagents and magnetic resolution agent are added in ELISA Plate or the tissue culturing plate, after reacting completely, ELISA Plate is advanced the magnetic separator from a thruster, within the 1min, can obtain separating.Then with ELISA Plate together with the separation vessel back-off, supernatant is poured out, and is placed on the clean filter paper, gently residual liquid is deducted.Extract ELISA Plate out from opposite side, can finish separation.Treat after the washing reusing when separate next time, convenient, convenient, and also applied widely.

Claims

1, a kind of immunoassay separates uses magnetic separator, it is characterized in that, comprise parallel placement, big plane stacks and clamps the last yoke plate (1) and the lower yoke plate (3) of the plate that is magnetic (2) therebetween, magnetic sheet (2), last yoke plate (1) and lower yoke plate (3) consistent size, each fixes the board-like framework of a strip (4) on the two sides of described lower yoke plate (3), the barb that has turning-in on the upper surface of the board-like framework of described strip (4), distance between the board-like framework of two strips (4) is 7-10.50cm, leaves the 1.0-2.0cm spacing between the lower surface of last yoke plate (1) and described barb.

2, the described immunoassay separation of claim 1 magnetic separator, it is characterized in that, described magnetic sheet (2) is the magnet plate of being made by ndfeb magnet, permanent ferrite, electromagnet, soft magnetic ferrite, aluminium nickel cobalt or SmCo magnetic material, or is a monoblock/several magnet plates that magnet plate is spliced.