Establishment of new and replacement World Health Organization International Biological Standards for human...

The complexity of the human interferon-alpha (IFN-alpha) family, with its multiple molecular forms and various biological activities, raises a number of scientific issues with regard to the biological standardisation of natural and recombinant IFN-alpha products. To address such issues and to achieve an appropriate biological standardisation of human interferon-alpha (IFN-alpha) preparations, the National Institute for Biological Standards and Control (NIBSC) of the United Kingdom (UK), in association with the Centre for Biologics Evaluation and Research (CBER) of the United States of America (USA), organised an international collaborative study, which was subsequently divided into two parts. Ninety-three participating laboratories from 29 countries worldwide participated in the first part of the study. They performed titrations on up to 15 different IFN-alpha preparations and one IFN-omega (omega) preparation in a variety of assays, including those based upon antiviral, antiproliferative, and other biological activities of IFN, and contributed raw data from these assays to NIBSC for analysis and calculation of relative activities. Analysis of data from this part of the study showed a greater than expected assay-dependent disparity between the relative activities of different IFN-alpha preparations. This disparity was found when only antiviral assays were considered and even when there were only small molecular dissimilarities between two otherwise closely related IFN-alpha preparations. The lack of assay independence and relative activity equivalence has indicated that a single biological potency standard for all IFN-alpha subtypes and mixtures would be inappropriate. Hence, individual, homologous standards, each with a separate unitage, were required for biological standardisation and potency determinations of individual IFN-alpha subtypes. At this stage, potency assignments to the IFN-alpha and -omega preparations included in the study were made as far as possible on the basis of comparison of antiviral activity with that of the 1st International Reference Preparation (IRP) for IFN, human leukocyte, 69/19. However, it was recognised that other standards had been used in assays to estimate potencies of widely available, current, therapeutic IFN-alpha products. Thus, to ensure the continuity of unitages already in use for IFN-alpha products, the second part of the study, which involved 12 members of the International Federation of Pharmaceutical Manufacturers Association (IFPMA), was carried out using for calibration of antiviral assays those IFN-alpha preparations that most closely matched manufacturers' products or that had been previously used for assay calibration by a manufacturer for a particular product. On the basis of data analysis from the second part of the study, potency assignments to the IFN-alpha preparations, as made in the first part of the study, were either left unchanged or changed to potency assignments that ensured as far as possible continuity with existing unitages. From among the IFN preparations evaluated, the following were recommended as the most suitable to continue or replace existing WHO international standards (IS) and have subsequently been formally established as WHO IS at the 51st meeting (October 1999) of the WHO ECBS: 83/514, 1st WHO IS for human IFN-alpha1 8000 international units (IU); 95/650, 2nd WHO IS for human IFN-alpha2a, 63,000 IU; 95/566, 2nd WHO IS for human IFN-alpha2b, 70,000 IU; 95/580, 1st WHO IS for human IFN-alpha2c, 40,000 IU; 95/572, 1st WHO IS for human IFN-alpha1/8, 27,000 IU; 94/786, 1st WHO IS for human IFN-alphaCon1, 100,000 IU; 94/784, 2nd WHO IS for human IFN-alpha (leukocyte), 11,000 IU; 95/574, 1st WHO IS for human IFN-alpha (leukocyte n3), 60,000 IU; 95/568, 2nd WHO IS for human IFN-alpha (lymphoblastoid n1), 38,000 IU; 94/754, 1st WHO IS for human IFN-omega, 20,000 IU. These WHO IS are available upon request to NIBSC.