Blanche Lane Potters Bar EN6 3QG Assuring the quality of biological medicines Tel: +44 (0)1707 641000 Template version: 18 February 2003 Fax: +44 (0)1707 646730 Page 1 of 4, 93/590 [email protected] http://www.nibsc.ac.uk

1st INTERNATIONAL STANDARD FOR PROTEIN S, PLASMA, HUMAN NIBSC code: 93/590 Version 02

1 THE STANDARD

The 1st International Standard for Protein S, Plasma, Human, consists of ampoules (code-labelled 93/590) containing approximately 1 ml aliquots of pooled fresh human plasma, freeze-dried. This standard has been assigned potencies for free and total Protein S antigen and for Protein S function.

2 CAUTION

2.1 This preparation is not for administration to humans.

2.2 A 'Material Safety Data Sheet' is included in these Instructions.

2.3 Each individual plasma donation and the final freeze-dried Standard was tested for HBsAg and for antibodies to HCV and HIV 1 and 2 and found to be negative. The Standard has also been tested and found negative for HCV RNA by PCR. However, as with all preparations of human origin, this material cannot be assumed to be free from infectious agents. Suitable precautions should be taken in the use and disposal of the ampoule and its contents. Such safety procedures probably will include the wearing of protective gloves and avoiding the generation of aerosols. Care should be exercised in opening ampoules, to avoid cuts.

3 USE OF THE STANDARD

Unopened ampoules should be stored at -20oC or below. Reconstitute the total contents of each ampoule of the Standard at room temperature with 1.0 ml of distilled water, using gentle shaking. Do not attempt to weigh out any portion of the freeze-dried material. Transfer the solution to a plastic tube and keep on ice during the assay. It is recommended that fresh ampoules are used for each assay, although the standard is sufficiently stable to be used over a 1 - 2 hour period.

4 ASSIGNED ACTIVITIES

The 1st International Standard was calibrated in an international collaborative study involving 16 laboratories for Total Protein S antigen, Free Protein S antigen and Protein S function by assay against fresh pools of normal plasma. The standard was established by the WHO Expert Committee on Biological Standardisation in October 1995. The following values have been assigned to the 1st International Standard:

Total Protein S antigen 0.90 IU per ampoule

Free Protein S antigen 0.90 IU per ampoule

Protein S function 0.90 IU per ampoule

Please note: Free Protein S antigen: in assay methods requiring PEG precipitation, the assigned potency for free Protein S antigen applies after PEG precipitation of the reconstituted standard.

National Institute for Biological Standards and Control 1470 A World Health Organization International Laboratory For Biological Standards

The Institute's UKAS accreditation relates only to our programme of testing for biological medicines. Full details can be found on our Schedule of Accreditation, which is available on request.

Blanche Lane South Mimms Potters Bar Hertfordshire EN6 3QG United Kingdom Assuring the quality of biological medicines Tel: +44 (0)1707 641000 Template version: 18 February 2003 Fax: +44 (0)1707 646730 Page 2 of 4, 93/590 [email protected] http://www.nibsc.ac.uk

Protein S function: the presence of the FV Leiden mutation may interfere with some assay methods for Protein S function; it is recommended that the specificity of assay methods for Protein S function should be validated.

5 BULK MATERIAL

The 1st International Standard was prepared in September 1993 from a pool of 4 litres of fresh plasma collected from 17 donors. Blood was collected into CPD/adenine contained in plastic packs, at a ratio of 450 ml blood to 67.5 ml of the anticoagulant. After centrifugation, the individual plasma donations were buffered by addition of HEPES (Hydroxyethylpiperazine-ethanesulphonic acid) to a final concentration of 0.05M, then centrifuged again and pooled.

6 DISTRIBUTION INTO AMPOULES

The pooled plasma was kept at 4oC throughout distribution into approximately 4,000 ampoules, then freeze- dried under conditions used for International Biological Standards (1). The mean liquid filling weight was 1.0101 gm (range 1.0051 to 1.0147 gm).

7 STABILITY

It is the policy of WHO not to assign expiry dates to international reference materials. Accelerated degradation studies have indicated that this standard is suitably stable, when stored at -20oC or below, for the assigned value(s) to remain valid until the standard is withdrawn or replaced. These studies have also shown that the standard is suitably stable for shipment at ambient temperature without any effect on the assigned value(s). NIBSC follows the policy of WHO with respect to its reference materials.

8 CITATION

In all publications (or data sheets for kits) in which this preparation is used as an assay calibrant, it is important that the title of the preparation, ampoule code number and the name and address of NIBSC are cited and cited correctly.

9 REFERENCE

1. Campbell P J (1974) J Biol Standardization, 2, 249-267.

10 Further information may be obtained from:

Division of Haematology National Institute for Biological Standards and Control Blanche Lane, South Mimms, Potters Bar, Hertfordshire, EN6 3QG, UK

Telephone: +44 (0)1707 641000 Fax: +44 (0)1707 646730

National Institute for Biological Standards and Control 1470 A World Health Organization International Laboratory For Biological Standards

The Institute's UKAS accreditation relates only to our programme of testing for biological medicines. Full details can be found on our Schedule of Accreditation, which is available on request.

Blanche Lane South Mimms Potters Bar Hertfordshire EN6 3QG United Kingdom Assuring the quality of biological medicines Tel: +44 (0)1707 641000 Template version: 18 February 2003 Fax: +44 (0)1707 646730 Page 3 of 4, 93/590 [email protected] http://www.nibsc.ac.uk

11. PRODUCT LIABILITY

i) Information emanating from NIBSC is given after the exercise of all reasonable care and skill in its compilation, preparation and issue, but is provided without liability in its application and use.

ii) This product is intended for use as a standard or reference material in laboratory work in relation to biological research, manufacturing or quality control testing of biological products or in the field of in vitro diagnostics.It is the responsibility of the user to ensure that he/she has the necessary technical skills to determine the appropriateness of this product for the proposed application. Results obtained from this product are likely to be dependent on conditions of use and the variability of materials beyond the control of NIBSC.

NIBSC accepts no liability whatsoever for any loss or damage arising from the use of this product, whether loss of profits, or indirect or consequential loss or otherwise, including, but not limited to, personal injury other than as caused by the negligence of NIBSC. In particular, NIBSC accepts no liability whatsoever for: - a) results obtained from this product; and/or b) non-delivery of goods or for damages in transit. iii) In the event of any replacement of goods following loss or damage a customer accepts as a condition of receipt of a replacement product, acceptance of the fact that the replacement is not to be construed as an admission of liability on NIBSC's behalf.

National Institute for Biological Standards and Control 1470 A World Health Organization International Laboratory For Biological Standards

The Institute's UKAS accreditation relates only to our programme of testing for biological medicines. Full details can be found on our Schedule of Accreditation, which is available on request.

Blanche Lane South Mimms Potters Bar Hertfordshire EN6 3QG United Kingdom Assuring the quality of biological medicines Tel: +44 (0)1707 641000 Template version: 18 February 2003 Fax: +44 (0)1707 646730 Page 4 of 4, 93/590 [email protected] http://www.nibsc.ac.uk

12. MATERIAL SAFETY SHEET

Physical properties (at room temperature) Physical appearance Solid Fire hazard None

Chemical properties Stable Yes Corrosive: No Hygroscopic Yes Oxidising: No Flammable No Irritant: No Other (specify) Contains material of human origin Handling: See caution, section 2

Toxicological properties Effects of inhalation: Not Known Effects of ingestion: Not Known Effects of skin absorption: Not Known

Suggested First Aid Inhalation Seek medical advice Ingestion Seek medical advice Contact with eyes Wash with copious amounts of water. Seek medical advice. Contact with skin Wash thoroughly with water.

Action on Spillage and Method of Disposal Spillage of ampoule contents should be taken up with absorbent material wetted with a virucidal agent. Rinse area with a virucidal agent followed by water. Absorbent materials used to treat spillage should be treated as biologically hazardous waste.

National Institute for Biological Standards and Control 1470 A World Health Organization International Laboratory For Biological Standards

The Institute's UKAS accreditation relates only to our programme of testing for biological medicines. Full details can be found on our Schedule of Accreditation, which is available on request.