CN1069970C - Method for activating test-tube of maleic acid anhydride-phenylethylene copolymer - Google Patents

Abstract

The invention discloses an activation method for a maleic anhydride-styrene copolymer test tube, belonging to the field of biochemistry. The problem of weak binding ability between the copolymer and the antibody and low activation rate existing in the previous activation method is solved. It is characterized in that the prepared maleic anhydride-styrene copolymer solution is coated on a polystyrene test tube, and sodium hydroxide solution is added into the coated test tube, and the solution in the tube is discarded after being placed at room temperature After washing and drying, the test tube of maleic anhydride-styrene copolymer is activated. The maleic anhydride-styrene test tube prepared by the above activation method has an activation rate increased by more than 30 times compared with that before activation; and it can be applied to solid-phase immunoassays of various antibodies and proteins, and has the advantage of a wide range of applications.

Description

A kind of maleic anhydride-activation method of styrene copolymer test tube

The invention discloses a maleic anhydride-styrene copolymer test tube activation method, which belongs to the field of biochemistry.

Maleic anhydride------styrene copolymer test tubes are mainly used in solid-phase immunoassays. Its activation refers to covalently binding antibodies or other proteins and polypeptides to polystyrene test tubes ( Plate) surface technology; that is, the introduction of active groups on the wall of the test tube, so that the protein or antibody can be covalently bound to it, so as to achieve the purpose of immobilizing the antibody. The currently known method is mainly to implant an amino polystyrene carrier on the test tube wall, and make the antibody covalently bond to the tube wall through a bifunctional group coupling agent, thereby immobilizing the antibody; the disadvantage is that the copolymer itself is not active. High, the ability to bind to proteins and antibodies is weak; it can no longer meet the actual needs.

The purpose of the present invention is to provide a kind of method is simple, the activation method of maleic anhydride------styrene copolymer test tube with strong binding ability between activated test tube and protein and antibody and wide application range.

The purpose of the present invention is achieved like this:

The present invention consists of the following steps: (1) preparation of maleic anhydride------styrene copolymer solution; after maleic anhydride and styrene are mixed in a molecular ratio of 2:1, benzoyl peroxide is added as a catalyst , the addition ratio is 0.2% of the weight of maleic anhydride; heat it in a water bath at 75-80°C, and immediately turn it into an ice-water bath after forming a viscous substance and boiling, and condense into a light yellow solid; after cooling, add the equivalent of benzene Butanone with 8 times the volume of ethylene is used to fully dissolve it, then add acetonitrile solution for dilution, and adjust the concentration of butanone to 15% to 20% to obtain a copolymer solution;

(2) Coating polystyrene plastic test tube: take the copolymer solution, pour it into the polystyrene test tube, and suck out the whole solution after standing for 10 seconds. Dry under conditions;

(3) Activation of maleic anhydride------styrene copolymer test tube: add 0.1-1.0mol/L sodium hydroxide solution into the coated test tube, leave it at room temperature for 6-24 hours, discard For the solution in the tube, wash the test tube with distilled water for 5 times, and dry it at 50°C, and the maleic anhydride-styrene copolymer test tube is activated.

The maleic anhydride-styrene copolymer test tube prepared by the above-mentioned activation method, due to the addition of sodium hydroxide solution, the copolymer is changed from hydrophobicity to hydrophilicity, which improves the ability to bind to proteins and antibodies; after activation The ability of the copolymer test tube to bind protein and antibody has increased from 4.0ug/tube before activation to 131ug/tube, which has increased the activation rate by more than 30 times; the prepared test tube can be stored at 4°C for 3 months, It can be stored for 3 years under the condition of ℃; it can also be applied to the solid-phase immunoassay of various antibodies and proteins such as polyclonal, monoclonal, and avidin, and has the advantage of a wide range of applications.

Claims (1)

1. a maleic anhydride---the activation method of polymers test tube is characterized in that being made up of following steps:

(1) maleic anhydride---the preparation of polymers solution: after maleic anhydride and styrene mixed by 2: 1 molecular proportions, add benzoyl peroxide and make catalyzer, additional proportion is 0.2% of a maleic anhydride weight; Place 75～80 ℃ of water-baths to heat, change over to immediately in the ice-water bath after thick material to be formed and the boiling, faint yellow solid congeals into; The cooling back adds the butanone that is equivalent to 8 times of volume of styrene makes it abundant dissolving, adds acetonitrile solution again and dilutes, and transfers to butanone concentration and reaches 15～20%, promptly gets copolymer solution;

(2) coating polystyrene test tube: get copolymer solution, inject the polystyrene test tube, leave standstill sucking-off complete soln after 10 seconds, the test tube tube wall after the coating leaves the layer of even co-polymer membrane, and room temperature condition is dry down;

(3) maleic anhydride---the activation of polymers test tube: the 0.1～1.0mol/L of the adding in vitro sodium hydroxide solution after coating, room temperature condition was placed 6～24 hours down, discard the solution in the pipe, with distilled water test tube is cleaned 5 times, under 50 ℃ of conditions, dry maleic anhydride---the styrol copolymer test tube promptly is activated.