GB2169312A

GB2169312A - A method for producing proteins

Info

Publication number: GB2169312A
Application number: GB08500007A
Authority: GB
Inventors: Peter Beaconsfield
Original assignee: BEACONSFIELD TINA
Current assignee: BEACONSFIELD TINA
Priority date: 1985-01-02
Filing date: 1985-01-02
Publication date: 1986-07-09
Also published as: GB8500007D0

Abstract

A method for producing proteins from living cells eg lymphocytes which method comprises irradiating the living cells with electromagnetic radiation of a frequency in the range of 10 x 10<6> Hz to 3 x 10<9> Hz.

Description

SPECIFICATION A Method for Producing Proteins The present invention relates to a method for producing proteins.

Proteins are complex polymers of amino-acids joined by peptide bonds. As presently performed the preparation of synthetic proteins from the constituent amino-acids is a complicated, expensive and inefficient procedure.

I have found that it is possible to induce living cells to produce increased amounts of proteins by irradiating them with electromagnetic radiation of specific energy levels.

Accordingly, the present invention provides a method of producing proteins from living cells which method comprises irradiating the living cells with electromagnetic radiation of a frequency in the range 10 x 106 Hzto 3 x 109 Hz.

Preferably the irradiation of the living cells is performed for periods of from 1 to 2 hours, followed by incubation of the cells for periods of for example, 2, 4, 6, 48, 72 or 120 hours.

Electromagnetic stimulation of the cells within the incubation medium was achieved using a signal generator, for example a Hewlett-Packard 5688A signal generator, through platinum needle or plate electrodes immersed in the incubation medium. For the higher ranges of frequency (i.e. above 300 MHz) coaxial cable was used.

The electromagnetic radiation may take the form of sine waves or square waves.

The following Examples illustrate the present invention.

Example 1 Globulin Production from Human Lymphocytes Standard, conventional procedures were used to isolate lymphocytes from whole blood taken from volunteers and used immediately. The isolated lymphocytes were suspended in an incubation medium RPMI 1640 (sold by Gibco) at a cell concentration of approximately 2 x 106 cells/ml. Glucose in a dose of 200 mg/100 ml was added to the incubation fluid. Where incubation was to last for more than 6 hours, 10% fetal calf serum was added to the fluid and an antibiotic incorporated to prevent infection in accordance with standard procedures for experiments involving relatively long periods of incubation. Samples (3, 4 or 5 ml) of incubation fluid carrying suspended lymphocytes were subjected to electromagnetic radiation at a frequency of 100 x 106 Hz at 25 A for 2 hours at 370C.The cells were irradiated in a chamber between opposed platinum plates, and the irradiation was performed using a Hewlett-Packard signal generator (Model Number 5686A) to generate sine waves at the chosen frequency. At the end of the period of irradiation, the samples were incubated for a further 6 or 12 hours. During incubation a flow of oxygen containing 5% CO2 was maintained.

Analysis of the samples for their content of y-globulins : IgG, IgM, and IgA was performed using standard measurement techniques such as those described in "Practical Immunology" by Hudson and Hay, Blackwells Scientific Publication, 1980. (See pages 233 - 239).

The following measurements were obtained: IgG lgM IgA (ng/100ml) (ng/100ml) (ng/100ml) Control Sample 1 1.9 < 1.0 < 1.0 Control Sample 2 1.6 < 1.0 < 1.0 Experiment Sample 1 10.1 1.8 2.8 Experiment Sample 2 8.3 2.1 1.9 Each sample has its own control, because human tissue is polymorphic.

The results demonstrate that the content of a-globulins is considerably increased in the irradiated samples relative to the control sample.

Radioactive studies demonstrated that the protein produced was newly synthesised, and not simply extruded by the cells into the medium. This new production was sustained for varying periods after stimulation had ceased.

Light and electron microscopy both gave evidence of increased cellular activity in the stimulated cells.

In all instances, stimulation of the same cell suspension can be repeated at intervals and further protein produced.

Example 2 Albumen Production from Liver Tissue Slices of liver tissue were taken either from otherwise healthy patients undergoing emergency abdomi nal surgery for repair of hepatic injury (usually resulting from a car accident) or from freshly killed Wistar rats. Each specimen weighed approximately 100 mg and was 1 mm thick. Two or three such specimens of liver tissue were suspended in 5 ml of the incubation medium described in Example 1, and then irradi ated in the same manner and with the same electromagnetic radiation as in Example 1. Experiments were started within 30 minutes of isolating the liver tissue.

The following measurements were obtained: Albumen in microgrammesilitre Control (1) 8.9 Stimulated (1) 10.9 Control (2) 8.7 Stimulated (2) 19.4 Control (3) 8.4 Stimulated (3) 15.4 Example 3 Insulin Production from Pancreas Tissue Example 2 was repeated, but using specimens of pancreas tissue instead of liver tissue. The specimens of pancreas tissue were taken from male mongrel dogs and were treated immediately in the usual manner to halt postmortem autolysis.

In each instance, newly synthesised insulin was recovered from the incubation medium.

Claims

1. A method of producing proteins from living cells which method comprises irradiating the living cells with electromagnetic radiation of a frequency in the range 10 x 106 Hz to 3 x 109 Hz.

2. A method according to Claim 1, wherein the irradiation is performed for 1 to 2 hours.

3. A method according to Claim 1 or 2, wherein following irradiation the cells are incubated for a period of up to 120 hours.

4. A method according to any one of Claims 1 to 3, wherein the electromagnetic radiation is in the form of sine waves.

5. A method according to any one of Claims 1 to 3, wherein the electromagnetic radiation is in the form of square waves.

6. A method according to any one of the preceding claims, wherein the protein generated is globulin.

7. A method according to any one of Claims 1 to 5, wherein the protein generated is albumen.

8. A method according to any one of Claims 1 to 5, wherein the living cells are human lymphocytes.

9. A method according to any one of Claims 1 to 5, wherein the living cells are liver cells.

10. A method according to any one of Claims 1 to 5, wherein the living cells are pancreas cells.

11. A method according to Claim 10, wherein the said proteins are isolated and recovered from the living cells.

12. A method of producing proteins from living cells substantially as described in any one of the foregoing Examples.

13. Proteins whenever prepared by a method as ciaimed in any one of the preceding claims.