World Intellectual Property Organization

Budapest Notification No. 144
Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure

Communication of the Republic of Italy Relating to the Acquisition of the Advanced Biotechnology Center (ABC)

The Director General of the World Intellectual Property Organization (WIPO) presents his compliments to the Minister for Foreign Affairs and has the honor to notify him of the receipt, on January 12, 1996, of a written communication, dated January 11, 1996, from the Government of the Republic of Italy, relating to the Advanced Biotechnology Center (ABC), indicating that the said depositary institution is located on the territory of the Republic of Italy and including a declaration of assurances to the effect that the said institution complies and will continue to comply with the requirements concerning the acquisition of the status of international depositary authority as specified in Article 6(2) of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure, done at Budapest on April 28, 1977, and amended on September 26, 1980.

The Advanced Biotechnology Center (ABC) will acquire the status of international depositary authority under the Budapest Treaty as from February 29, 1996, the date of publication of the said communication in the February 1996 issue of Industrial Property and Copyright/La Propriété industrielle et le Droit d'auteur.

February 14, 1996


Text of the Written Communication of the Government of the Republic of Italy, dated January 11, 1996, Relating to the Acquisition of the Status of International Depositary Authority by the Advanced Biotechnology Center (ABC)

[Original: French; English translation prepared by the International Bureau of WIPO]

COMMUNICATION

The Permanent Mission of Italy to the United Nations Office and other international organizations in Geneva presents its compliments to the World Intellectual Property Organization and has the honor to submit a formal request for recognition of the status of international depositary authority, under Article 7 of the Budapest Treaty of April 28, 1977, for the Advanced Biotechnology Center, whose headquarters are in Genoa.

The Italian Patent and Trademark Office of the Ministry of Industry has given assurances in that respect that the Center possesses the necessary functional qualities and that it satisfies the conditions of competence, impartiality and structural and professional capacity referred to in detail in Article 6(2) of the Budapest Treaty.

Pursuant to Rule 3 of the Regulations under the Budapest Treaty with regard to acquisition of the status of international depositary authority, the Permanent Mission of Italy wishes to submit herewith information on the Management Consortium of the Advanced Biotechnology Center and on the collection of cell lines.

The Permanent Mission also transmits herewith a letter of presentation and an explanatory brochure* on the above-mentioned Center.

(* Not reproduced here)


INFORMATION ON THE MANAGEMENT CONSORTIUM OF THE ADVANCED BIOTECHNOLOGY CENTER AND ON THE COLLECTION OF CELL LINES

I. Characteristics of the Depositary Institution

A. General Information

1. There is at present in Italy no international depositary authority authorized under the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.

2(a) The Management Consortium of the Advanced Biotechnology Center (ABC) within which the Interlab Cell Line Collection (ICLC) operates has its headquarters at Largo Rossana Benzi, 10, 16132 Genoa, Italy.

2(b) The Management Consortium of the Advanced Biotechnology Center was set up on April 27, 1993, and has been in operation since November 1994.

2(c) The ICLC cell line bank within the Advanced Biotechnology Center accepts the deposit of human and animal cell lines and samples of cancerous tissue.

It is also planned to increase activities and deposit services within the microorganism and animal embryo sectors on the basis of collaboration between ICLC and other services operating within the Advanced Biotechnology Center.

The Management Consortium of the Advanced Biotechnology Center is waiting to be recognized as an international depositary authority (under Article 6 of the Budapest Treaty) in order to accept, in this initial phase, cell lines and, subsequently, also microorganisms for the purposes of patent procedure.

2(d) ICLC uses the following languages:

- Italian;
- English;
- French;
- German.

B. Legal Status and Funding

3(a) The Management Consortium of the Advanced Biotechnology Center is a private establishment.

It is a consortium whose founders are the University of Genoa and the National Institute for Cancer Research.

3(b) Supervision of the Management Consortium of the Advanced Biotechnology Center is guaranteed by the appointment of public servants.

The Supervisory Council of the Consortium is, indeed, composed in majority of representatives of ministries and administrative organs: the Ministry of Health, the Ministry of University Matters and Scientific and Technological Research, the Higher Institute of Health; the National Research Council. The Board of Auditors of the Consortium is composed exclusively of public servants representing the following ministries: the Ministry of Finance, the Ministry of University Matters and Scientific and Technological Research, the Ministry of Health.

3(c) Funding of ABC is provided by:

- public and private research contributions;
- transfers from the organs that compose the Consortium;
- the proceeds of research activities and scientific services.

C. Staffing

4. 231 persons were employed by ABC on December 31, 1994. Seven persons were specifically in the ICLC.

5. They include four persons holding a university degree.

6. The ICLC organigram is as follows:

- 1 laboratory director,
- 3 graduates,
- 2 technicians,
- 1 secretary.

D. Premises of the Depositary Institution

7(a) ABC occupies a surface of approximately 16,000 m2 of which 10,000 m2 are devoted to research laboratories.

Within ABC, the structures and support services for biomedical research are available and already operational.

The ICLC bank, operating within the Center, occupies 110 m2 .

7(b) The following principal equipment is available at ICLC for use in current activities for the deposit of human and animal cell lines:

AREA BL3
Electronically controlled autoclave

Quarantine chamber (cell lines that are not tested for microplasmas)
Laminar flow dome
CO2 incubator

Sterile chamber
Laminar flow domes
CO2 incubator
Inverted microscope
Thermostatic bath

Laboratory
Chemical dome
Laminar flow dome
Frozen bench centrifuge
Electrophoresis feeder
Apparatus for isoenzyme testing
pH-meter

Store
freezer, -20°
2 refrigerators with freezer

Freezing Chamber
Recipient for liquid nitrogen (8,000 ampoules) with an alarm system and an automatic filler system
Recipient for liquid nitrogen (750 ampoules)

Office
Personal computer 386

Equipment shared with other laboratories
Fluorescence microscope
Freezer, -80°, 368 liters
Thermal cycler

A list of equipment and a plan of the ABC laboratories which also work in the fields of microorganisms and animal embryos will be supplied when the collaboration agreements have been defined.

II. Activities

8. The ICLC bank currently accepts for deposit, pending extension of its activities, human and animal cell lines and tissue samples.

9. The ICLC bank is currently able to conserve approximately 400 cell lines (20 ampoules for each line).

10(a) During the year in which the ICLC bank has been in operation (since November 1994) approximately 50 deposits have been made.

10(b) No deposit made so far has been in relation to patent matters due to lack of recognition as an international depositary authority.

10(c) Thirteen deposits have been made by depositors resident in Italy.

11. The ICLC bank currently publishes a catalogue, either on paper or on magnetic medium, of the cell lines stored, which will be supplemented with all the information on the material subsequently stored for patent purposes.

12(a) The ICLC bank registers the information on the cell lines in the cell line database already operative in the Advanced Biotechnology Center.

The database was set up under the Interlab project and contains the data on: identification, origin, typology, characterization, quality control and culture characteristics of some 3,000 cell lines available in various Italian and foreign laboratories. The CLDB database was set up using the Oracle relational database management system but can be interrogated on-line over the Internet even on the basis of other network software (Gopher, World Wide Web, Wais) and cell lines of interest may be requested by e-mail. The news service of the database is used to make information available on new lines included in the bank, relevant congresses, etc. It is also planned to periodically print general or specialized catalogues containing detailed information on the cell lines available in the bank.

12(b) The Management Consortium of the Advanced Biotechnology Center undertakes to provide an information service for matters of customs, quarantine and postal regulations in Italy.

III. Deposit Procedures for Microorganisms

A. Acceptance for Deposit of Cell Lines

13. The Management Consortium of the Advanced Biotechnology Center requires the following conditions to be complied with for acceptance for deposit:

(a) frozen samples of cell lines--at least 12 ampoules for each line, each containing 2-5 times 106 cells--are to be sent to (the Management Consortium of the Advanced Biotechnology Center--ICLC Bank) in expanded polystyrene recipients containing a quantity of dry ice ensuring 48 hours autonomy at room temperature.

(b) The information to be supplied by the depositor is:

a written declaration signed by the depositor and containing:

- a statement that the deposit is made under the Budapest Treaty and a commitment not to withdraw it during the period of time set out in Rule 9.1 of the Regulations,

- the name and address of the depositor,

- a detailed description of the conditions to be respected for cultivating the cell line, for conserving it and for checking its viability,

- the name of the cell line and any identification code (number, symbol, etc.) given by the depositor,

- a statement of the properties of the cell line that are dangerous or may be dangerous for health or the environment or a statement that the depositor is not aware of any such properties. The depositor undertakes to inform ICLC promptly of any new information he may obtain on properties of the deposited line which may modify the declarations already made.

Give in the written declaration the scientific description and origin of the deposited cell line.

14. The form* drawn up by the ICLC Bank containing the information to be supplied by the depositor at the time of deposit is attached.

(* Not reproduced here)

B. Conservation

15(a) Described are the storage and quality control methods for deposited cell lines in order to ensure viability and lack of contamination.

The viability of cell lines for patent procedure is checked on arrival by defreezing and cultivating one of the deposited ampoules.

The technique uses a vital stain (trypan blue) that is to say which can penetrate and therefore stain only the dead cells. The coloration is specific for the first ten minutes only.

The cells are then counted and viability is assessed, that is to say the percentage of living cells in relation to the dead cells. The cell line culture is then spread in a category BL3 sterile chamber under a laminar flow dome. The presence of contamination by bacteria, fungi and yeasts in the cultures is often easily identifiable for the evident effects by either macroscopic inspection (cloudiness of the culture medium) or by microscope.

Most contaminations can be avoided by proper laboratory practice:

(1) sterility test of the prepared culture mediums; (2) use of culture mediums devoid of antimicrobials; (3) training of staff to work in a sterile atmosphere; (4) attention to possible sources of contamination in the laboratory (refrigerators, thermostatic baths, use and correct maintenance of laminar flow domes by careful cleaning and periodic checking of filters).

The contamination of lines by microplasms assumes a special importance that is not directly evident in the cultures and may impair the normal structural and functional characteristics of the cell.

The precautions to be taken to avoid culture contamination are: not to use antibiotics which suppress but do not kill the microplasms; test the lines regularly for microplasms. The growth of microplasms in cultures can be determined by a direct method (culture test on a specific agar) or an indirect method (biochemical analysis coloration). Only the appropriate use of differing methods can exclude contamination; the direct method is very sensitive but does not permit the identification of microplasm strains responsible for all contaminations; the indirect methods are rapid but less sensitive. It should be remembered that cell lines are a potential source of viruses and that some of those have not yet been identified. Even the vital serum that is not tested may be a source of contamination.

Once again, working under sterile conditions and careful checking of the culture mediums can avoid the spread of viruses in the cultures. It is important for the protection of the operative that the presence in lines of viruses such as HTLV-I et II, HIV and BVDV be notified.

The line freezing processes must be carried out with care in order to conserve culture viability. The quality of the nitrogen recipients, the periodic checking of the alarm systems and the general organization of the area in which the recipients are located are very important.

It is very important to use more than one recipient so that a security stock of ampoules for each line is conserved. The preparation and despatch of material

for distribution must be rapid to avoid lines remaining at temperatures that are not recommended.

(b) The security measures used to reduce to a minimum the risk of losing deposited microorganisms are described.

Since it is not always convenient to keep cell lines under culture at all times for various reasons (possibility of contamination, or cross contamination, genetic alterations, limited reproduction capacity, cost) the production of two homogeneous crioconserved quotas of each culture is a first step to ensuring the supply of cells over time. In most laboratories, a stock is used and is reconstituted from the last available ampoule. However, in this way the characteristics of the line tend to change and with the number of passages the line is sure to change and the cells with which one works become very distant from the original line.

The correct storage practice for a line, on the contrary, consists in creating a master bank from the original deposit (10-20 ampoules) and a working bank or distribution stock (30-50 ampoules). In this way all the cells distributed will derive from the same original stock.

C. Fees

16. The amount of the fees due (at the time of) making the deposit:

(a) 2 million lire for conserving the cell lines/hybridoma;
(b) 100,000 lire for issuing a viability statement;
(c) 250,000 lire plus the cost of despatch for providing a sample;
(d) 200,000 lire for any notification activities or requests for authorization

from the competent authorities, except for the tariffs and/or fees and despatch costs.

17. The amount of the fees does not depend on the nationality or the domicile of the depositor, the authority or the natural or legal person requesting issue of a certificate of viability or the supply of a sample.

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