JPS61264249A - Production of vital material immobilized film - Google Patents

Abstract

PURPOSE:To improve the reproducibility of sensor output by imposing a spacer bored preliminarily with a through-hole on a support of a substrate transmittable film, filling a monomer soln. contg. a vital material in the through-hole of the spacer and polymerizing the same thereby removing the spacer. CONSTITUTION:The substrate transmittable film 2 having 24Angstrom pore size and 50mum thickness is adhered tightly onto a glass plate 1 having a flat surface and the spacer 3 bored preliminarily with the through-hole made coincident with the diameter of a vital material immobilized film 6. The monomer soln. 4 contg. the vital material which is enzyme or microorganisms is dropped into the through-hole of the spacer 3 so as to be filled in the through-hole and is polymerized. The hole is capped with a glass plate 5 and the light of a fluorescent lamp is irradiated thereto to complete the polymn. in ths stage. The plate 5 and the spacer 3 are removed and the immobilized film 6 is formed. The monomer is an acrylamide or 2-hydroxyethyl methacrylate. The reproducibility of the sensor output is improved and the response time is reduced.

Description

[Detailed description of the invention] [Summary] Base! A spacer with pre-drilled through holes is placed on top of the permeable membrane support, and a monomer solution mixed with a biological substance is filled into the through holes of the spacer and polymerized. After that, the spacer is removed and the spacer with the support is removed. Manufacture a biological material immobilization membrane.

[Industrial application field]

The present invention relates to a biological substance immobilized membrane, particularly to a method for producing an immobilized membrane that maintains its shape well and has a predetermined size.

[Conventional technology and problems]

The membrane that immobilizes enzymes and microorganisms is made of gel such as polyacrylamide, but it is not only difficult to adjust the shape and thickness of gel, but also has low mechanical strength, so it is necessary to make the membrane thicker so that it retains its shape. There is a need to. Polymer membranes containing such biological substances are used by attaching them to galvanic or polarographic oxygen electrodes, but if the membrane is thick, the reproducibility of the sensor output is low and the response time is significantly long. .

[Means for solving problems]

The above problem lies in the method for producing a polymer membrane containing biological substances, in which a spacer with pre-drilled through holes is placed on a substrate-permeable membrane support, and a monomer solution mixed with biological substances is prepared. This problem can be solved by a method for manufacturing a biological material-fixed membrane, which is characterized in that the spacer is removed after filling the through-hole of the spacer with polymer and polymerizing it.

The biological material can be an enzyme or a microorganism.

The substrate permeable membrane is conveniently a dialysis membrane or an ultrafiltration membrane.

The monomer is preferably acrylamide or 2-hydroxyethyl methacrylate.

〔Example〕

As shown in FIG. 1, (a) a dialysis membrane 2 with an average pore diameter of 24 and a thickness of 50 μm is placed on a glass plate 1 having a flat surface.
A spacer 3 made of a fluororesin membrane with a thickness of 50 μm and having a through hole of 5 m in diameter in accordance with the diameter of the intended fixed membrane was placed thereon. (b) 10 μg of glucose oxidase, 1 g of acrylamide monomer and N as crosslinker.

N'-methylenebisacrylamide 50μ pH 7.4
Dissolved in 5 ml of phosphate buffer, and further added 4 μl of potassium persulfate and 0.2 μl of riboflavin as polymerization initiators.
Drop the prepared solution 4 containing ug into the through hole to fill it (
C) Cover with a glass plate 5 and irradiate with fluorescent light from above. Polymerization was completed after about 30 minutes, and the glass plate 5 and spacer 3 were removed to form a polymer film with a thickness of 50 μm and a diameter of 5 mm on which glucose oxidase was immobilized.

C. Effects of the Invention] According to the present invention, a biological substance fixed membrane having a predetermined thickness and size is obtained, and since it has a support, it has high strength and can be easily attached to a galvanic cell type or polarographic type oxygen electrode. It has the advantages of being easy to wear, having high reproducibility of sensor output, and short response time.

[Brief explanation of the drawing]

FIG. 1 is a diagram showing the manufacturing process of a biological material-fixed membrane according to the present invention. 1.5...Glass plate, 2...Substrate permeable membrane, 3
...Spacer, 4. Monomer solution containing biological material, 6. Biological material immobilized membrane.

Claims (1)

[Claims] 1. A method for producing a polymer membrane containing a biological material, in which a spacer with a through hole formed in advance is placed on a substrate-permeable membrane support, and a polymer membrane containing a biological material is produced. 1. A method for producing a biological material-immobilized membrane, which comprises filling a through-hole of a spacer with a polymer solution and polymerizing it, and then removing the spacer. 2. The production method according to claim 1, wherein the biological substance is an enzyme or a microorganism. 3. The manufacturing method according to claim 1, wherein the substrate permeable membrane is a dialysis membrane or an ultrafiltration membrane. 4. The manufacturing method according to claim 1, wherein the monomer is acrylamide or 2-hydroxyethyl methacrylate.