Pharmeuropa bio最新文献

The European Pharmacopoeia (Ph. Eur.) monograph Human tetanus immunoglobulin (0398) gives a clear outline of the in vivo assay to be performed to determine the potency of human tetanus immunoglobulins during their development. Furthermore, it states that an in vitro method shall be validated for the potency estimation. Since no further guidance is given on the in vitro assay, every control laboratory concerned is free to design and validate an in-house method. At the moment there is no agreed method available. The aim of this study was to validate and compare 2 alternative in vitro assays, i.e. an enzyme-linked immunoassay (EIA) and a toxoid inhibition assay (TIA). The potency of 2 tetanus immunoglobulin preparations (Product 1, Product 2) was estimated against the WHO International Standard for tetanus immunoglobulin, using the tetanus EIA and TIA. The coefficient of variation (CV) to characterise the assay precision was 3.2% (EIA) and 3.6% (TIA), and the corresponding CV for intra-assay variation was 4.7% (EIA) and 5.5% (TIA). Using a spiking procedure, the 2nd part of the experiment investigated recovery of a known anti-tetanus potency. The recovery of samples spiked with defined amounts of reference preparation ranged from 104 112% (EIA) and 114 125% (TIA) respectively, resulting in a mean bias of 2.2 IU/ml (95% confidence interval (CI): -1.1-5.4 IU/ml, EIA) and 5.8 IU/ml (95% CI: 1.4 10.2 IU/ml, TIA). Good agreement was observed between the in vivo and in vitro assay results: the relative potency results of the EIA and TIA as compared to those of the in vivo assay performed by the manufacturers of the 2 tetanus immunoglobulins were for the EIA in the range of 104+/-10% for Product 1 and 100+/-6% for Product 2, and for the TIA in the range of 107+/-6% for Product 1 and 100+/-7% for Product 2. Tetanus EIA and TIA are suitable quality control methods for polyclonal tetanus immunoglobulin, which can be standardised in a quality control laboratory using a quality assurance system. In a collaborative study it will now be evaluated whether the validated methods can be proposed as common in vitro batch potency assays for replacement of the in vivo mouse assay.

A study was carried out by the European Directorate for the Quality of Medicines (EDQM) as part of the joint Biological Standardisation Programme of the Council of Europe and the European Commission with the aim to establish replacement batches of the European Pharmacopoeia (Ph. Eur.) human immunoglobulin Biological Reference Preparation (BRP) batch 2. Twenty-eight laboratories participated in this study. The suitability of the candidate reference preparations to serve as working references in the tests for distribution of the molecular size, anticomplementary activity and Fc function, in accordance with the specifications of the Ph. Eur. monographs Human normal immunoglobulin for intravenous administration (0918), Human normal immunoglobulin (0338) and Anti-T lymphocyte immunoglobulin for human use, animal (1928) was demonstrated. The candidates were therefore established as human immunoglobulin BRP batch 3 and Human immunoglobulin (molecular size) BRP batch 1. The prescribed use of the latter BRP is limited to the test for distribution of molecular size.

European Pharmacopoeia (Ph. Eur.) general chapter 2.6.7. Mycoplasma requires for the culture test reference strains of mycoplasma field isolates with fewer than 15 passages for validation and run control and in the test for inhibitory substances. Low passage field isolates of 5 mycoplasma strains (Mycoplasma hyorhinis, Mycoplasma synoviae, Mycoplasma fermentans, Mycoplasma orale and Acholeplasma laidlawii) have been prepared for this purpose and a small scale collaborative study involving European laboratories was carried out to confirm the suitability of the material for the intended purpose. Strains were prepared as 1 ml samples in frozen format and are stored below -60 degrees C. Each laboratory determined a titre for the material on their in-house media. A secondary part of the study also compared the growth of prediluted samples on the different culture media. Results of the study confirm that the material is suitable for use as a biological reference preparation (BRP) and an estimated titre has been provided for each strain based on the results of the study. It was noted that differences in the culture media used in the different laboratories did not have a detrimental effect on titre estimation. The estimated titre is intended as a guide for users to validate the use of the reference material in house. The candidate BRPs were adopted by the European Pharmacopoeia Commission on June 28, 2006 and are available for use from EDQM. A revision to chapter 2.6.7, including reference to the use of nucleic acid amplification techniques (NAT) was also adopted in June 2006 and will appear in the European Pharmacopoeia version 5.8 in January 2007 and come into force the 1st of July 2007. While it was not part of the study a number of participants also performed in-house NAT assays on the study material. Preliminary findings from these studies are presented.

For the potency assay of human coagulation factor VII concentrate preparations according to the European Pharmacopoeia (Ph. Eur.) a reference preparation calibrated in International Units (IU) is needed. Currently, the 1st International Standard (97/592, potency: 6.3 IU/ampoule) but no Ph. Eur. reference preparation is available. A collaborative study was run to calibrate a candidate Ph. Eur. Biological Reference Preparation (BRP) for human coagulation factor VII concentrate against the 1st International Standard; the BRP is intended to be used as working standard. A candidate BRP batch 1 was produced from a plasma-derived human factor VII concentrate preparation available on the European market. It fulfilled the requirements of a BRP with regard to precision and homogeneity of fill, residual water content and stability. In addition, the content of activated factor VII was low. Sixteen laboratories from 9 countries participated in the collaborative study. The potency of the candidate BRP was determined using the participants' chromogenic assay based on the Ph. Eur. and their in-house clotting assay, if available. The statistical model used for analysis of the results from most laboratories was the maximum likelihood of the parallel line model following a logarithmic transformation of the responses. In the chromogenic assay, a potency estimate of 8.2 IU/vial (+/-3.7%) was obtained for the candidate BRP. Results from the clotting assay were lower and less homogenous (6.7 IU/vial+/-11.6%). The results from the collaborative study showed that the candidate BRP is suitable as a reference standard for the chromogenic assay according to the Ph. Eur. It was adopted by the Ph. Eur. Commission in March 2006 as official Ph. Eur. BRP for this purpose.

An international collaborative study was organised to establish a European Pharmacopoeia (Ph. Eur.) Biological Reference Preparation (BRP) and United States (US) Food and Drug Administration (FDA) reference preparation for the test for anti-D (anti-Rho) antibodies in human normal immunoglobulin for intravenous administration (IGIV). A candidate positive control (IGIV+anti-D) and negative control IGIV were compared to corresponding World Health Organization (WHO) International Reference Reagents using a direct haemagglutination reference method. Sixteen (16) laboratories participated in the collaborative study. Further to completion of the study, the materials assayed in the study were granted the status of Ph. Eur. and US FDA reference preparations for controlling the levels of anti-D in IGIV.

A project was run for the establishment of replacement batches of the European Pharmacopoeia (Ph. Eur.) Somatropin Chemical Reference Substance (CRS) batch 1. Twenty two laboratories from 16 countries took part in a collaborative study aimed at demonstrating the suitability of the candidate reference preparations to serve as working references in the tests for identification by peptide mapping and capillary electrophoresis (CE); related proteins, dimers and related substances of higher molecular mass; charged variants distribution; and/or for the assay of somatropin, as performed in accordance with the specifications of the current Ph. Eur. monographs 0950 Somatropin bulk solution, 0951 Somatropin and 0952 Somatropin for injection. Further to the completion of the study the Ph. Eur. Commission adopted one candidate in March 2006 as somatropin CRS batch 2 (with an assigned content of 1.69 mg somatropin monomer per vial) and the second one in June 2006 as somatropin/desamidosomatropin resolution mixture CRS batch 1 (prescribed use of the latter standard is restricted to the test for related proteins).

The study is a contribution to the EDQM's efforts to meet some of the expectations of the 3 Rs: Replacement, Reduction and Refinement of animal assays as proposed by Russell and Burch in 1959 and adopted by the European Union in 1986, and specifically to validate alternative assays to replace, for batch-release purposes, the European Pharmacopoeia (Ph. Eur.) in vivo direct challenge procedures for the potency determination of diphtheria toxoid vaccines. The study results may be used in support of the replacement of the multi-dilution direct challenge procedures in different animal models by a single dilution serology test, where appropriate, and to use sera from the same animals for potency testing of several components in combined vaccines. With regard to the latter, the present study explores the possibility of testing both diphtheria and tetanus toxoid potencies using serum from the same animals.

The discontinuation of the Auszyme kit used by vaccine manufacturers and national control laboratories to determine the Hepatitis B surface antigen (HBsAg) content of hepatitis B vaccines has led GlaxoSmithKline (GSK) to develop an alternative inhibition ELISA method. Validation of this ELISA was performed according to The International Conference of Harmonization and reproducibility was assessed in a feasibility study with four Official Medicines Control Laboratories (OMCLs). The dose response curve demonstrated linearity (R2>0.99) in the range of 60-360 ng/ml HBsAg. The repeatability (CV<7%), intermediate precision (CV<10%) and accuracy (91-113% recovery) were similar to the Auszyme method. The commercial antibodies used in the assay were shown to contain antibodies that bind to a protective epitope of HBsAg and the specificity of the method for HBsAg was demonstrated. There was a good concordance with the Auszyme method, although the ELISA yielded higher results (25.3 vs. 24.4 micro.g/ml for Engerix-B (n=64), 28.9 vs. 27.0 micro.g/ml for Twinrix (n= 69) and 25.5 vs. 21.6 micro.g/ml for Infanrix penta (n=62)). The method was successfully transferred to the four OMCLs. It has been demonstrated that the ELISA is suitable for its intended purpose with hepatitis B-containing vaccines from GSK and thus could be used for these vaccines by national control laboratories and authorities. Further validation studies should focus on the use of this ELISA with vaccines from other manufacturers.

Upon suggestion of the French Official Medicines Control Laboratory, a collaborative study was initiated by the European Directorate for the Quality of Medicines with the goal of calibrating the candidate European Pharmacopoeia biological reference preparation (Ph. Eur. BRP) for anti-vaccinia immunoglobulin batch 1 in International Units (IU) against the 1(st) British standard (anti-smallpox serum). The candidate BRP batch 1 was obtained by lyophilising a pool of four plasma samples obtained from one donor who was multi-vaccinated with smallpox vaccine (Lister strain) and who had relatively high titres of neutralising anti-vaccinia antibodies. The plasma complied with the requirements of the Ph. Eur. monograph Human plasma for fractionation. For the candidate BRP the precision of fill and the residual moisture after lyophilisation comply with the requirements for biological reference preparations. The stability of the material was shown to be satisfactory for the intended purpose in an accelerated degradation test. Eight laboratories participated in the study. Two samples had to be assayed (candidate BRP batch 1 and 1(st) British standard). All participants were requested to test the samples using a common method (plaque reduction neutralisation) that had been validated beforehand, and their own in-house anti-vaccinia immunoglobulin titration method. From the raw data returned, the potency of the candidate BRP was calculated in IU/ml using the parallel lines method. The precision (intra-assay variation), repeatability (intra-laboratory variation) and reproducibility (inter-laboratory variation) were assessed. All laboratories used the Lister strain of vaccinia virus for the plaque reduction neutralisation assay. For laboratories using cell-adapted vaccinia virus, the results were satisfactory regarding intra-assay variability, intra-laboratory variability and inter-laboratory variability. For laboratories using vaccinia virus produced on animals, results were less satisfactory. The study suggests that the candidate BRP batch 1 is suitable as a reference preparation for the potency assay of vaccinia immunoglobulin by the plaque reduction neutralisation method, using cell-adapted vaccinia virus. For this purpose, a potency of 23 IU/vial could be assigned to the candidate BRP. Based on the results of the stability testing, storage of the reference material at -20 degrees C and shipment on ice is recommended. Furthermore, it is recommended to monitor the potency of the reference material once per year. The candidate material was adopted as Ph. Eur. BRP at the Ph. Eur. Commission session in March 2005.

A feasibility study was organised to determine the possibilities for development of a common in vitro assay for determination of D-antigen content in inactivated poliomyelitis vaccines (IPV). 3 different methods were tested on a selection of non-combined IPV vaccines from the European market. The results of this preliminary study suggest that for vaccines with a similar strain composition similar results would be achieved regardless of which of the three methods was used. Nevertheless, for one vaccine with a slightly different strain composition the results obtained depended on which method was applied. This highlights the need to take into account the strain composition in any future development of a common method. The study also highlighted the importance of standardising the statistical approach to analysis of results, since one laboratory obtained different sets of results by applying different statistical analysis to the same raw data. While no immediate need was seen for a large collaborative study to establish a common method, participants encouraged the idea of further study, in particular with respect to the different strain compositions. Adaptation of a common method will also require further analysis of the needs for combined vaccines, including the steps and conditions for de-sorption.