Pharmeuropa bio & scientific notes最新文献

A study was organised by the European Directorate for the Quality of Medicines & HealthCare (EDQM, Council of Europe) to establish a replacement batch for the current European Pharmacopoeia (Ph. Eur.) Human Anti-D immunoglobulin Biological Reference Preparation (BRP) batch 1 whose stocks were dwindling. The BSP176 study was run in the framework of the Biological Standardisation Programme (BSP) of the Council of Europe and the European Union (EU) Commission. In view of the current stocks, the difficulties in sourcing suitable starting materials and the time required to calibrate a replacement batch with the 3 different methods described in Ph. Eur. chapter 2.7.13. Assay of human anti-D immunoglobulin, it had been decided as an interim measure to establish BRP batch 2 (BRP2) using ampoules already qualified for use as the World Health Organization (WHO) 3^rd International Standard (IS) for anti-D immunoglobulin (16/332) and obtained from MHRA. For this purpose, a verification study has been performed at the EDQM laboratory on the candidate BRP (cBRP) material, i.e. ampoules of the 3^rd IS for anti-D immunoglobulin received at EDQM, to be used as BRP2. The study showed that the anti-D estimated potency of the cBRP, WHO 3^rd IS for anti-D Immunoglobulin, relative to BRP1 which is equivalent to the WHO 2^nd IS, was consistent with the value of 297 IU/ampoule assigned during the establishment study for the 3^rd IS and in continuity with BRP1. Stability was confirmed by checking the potency of ampoules stored at the recommended storage temperature (i.e. -20°C) against ampoules stored at -70°C since the establishment. The molecular-size distribution profile of the cBRP was also similar to the one obtained during the establishment study, again confirming the stability. All these data confirmed the good stability of the WHO 3^rd IS for anti-D Immunoglobulin in line with the stability study performed during its establishment and confirmed its suitability for use as BRP2. In October 2024, the Ph. Eur. Commission officially adopted the candidate batch as Ph. Eur. Human Anti-D immunoglobulin BRP batch 2 with an assigned potency of 297 IU/ampoule. The activity of the new batch of Ph. Eur. Anti-D immunoglobulin BRP will be regularly monitored.

An international collaborative study was organised by the European Directorate for the Quality of Medicines & HealthCare (EDQM, Council of Europe) to calibrate replacement batches for the current European Pharmacopoeia (Ph. Eur.) Prekallikrein Activator (PKA) in albumin Biological Reference Preparation (BRP) whose stocks were dwindling. The study was run in the framework of the Biological Standardisation Programme (BSP) of the Council of Europe and the European Union (EU) Commission. Twenty-four laboratories from official medicines control authorities and manufacturers in Europe and outside Europe took part in the study. Three candidate replacement batches were produced with albumin solutions artificially spiked with a PKA concentrate to increase their PKA level. Participants were requested to evaluate the candidate batches against the 3rd World Health Organization (WHO) International Standard (IS) for Prekallikrein activator in albumin (16/364) using their routine assay method. The Ph. Eur. PKA in albumin BRP batch 7 (BRP7) was also included in the test panel to ensure the continuity of the consecutive BRP batches. The 3 candidate replacement batches were considered suitable for their intended use as BRPs. The study confirmed the stability of the PKA content of the current BRP7. Thermal stress study on the candidate batches confirmed the stability of their PKA activity. In December 2023, the Ph. Eur. Commission officially adopted the 3 candidate batches as Ph. Eur. PKA in albumin BRP batches 8, 9 and 10 with assigned potencies of 37 IU/vial, 33 IU/vial and 34 IU/vial, respectively. The activity of the 3 new batches of Ph. Eur. PKA in albumin BRP will be regularly monitored.

Recombinant adeno-associated viruses (AAV) are widely used as gene therapy vectors in human gene therapy. Reliable and accurate quantification of the physical particle titre is one of the parameters to be determined for precise dosing, which is of critical importance for the patients. In this report, we describe the validation of an enzyme-linked immunosorbent assay (ELISA) for the determination of the total AAV-2 physical particle titre through an international collaborative study organised by the Gene Therapy Working Group of the General European Official Medicines Control Laboratories Network co-ordinated by the European Directorate for the Quality of Medicines & HealthCare (EDQM), which aims to develop and validate standard analytical methods for the quality control of gene therapy products. The method is based on a classical sandwich ELISA which uses monoclonal antibodies specific for a conformational epitope only present on the assembled capsid. The AAV-2 Reference Standard Stock Material was used as reference preparation. This study allowed validation of the sensitivity, accuracy and reproducibility of the method and definition of its working range. In addition, specificity and discriminatory capacity towards degraded preparations was also demonstrated.

Tetanus vaccines for human and veterinary use are produced by formaldehyde-induced inactivation of tetanus neurotoxin (TeNT) purified from Clostridium tetani cultures. Due to the high morbidity caused by exposure to TeNT it is essential that the quality control of tetanus vaccines includes testing for absence of tetanus toxin as prescribed by European Pharmacopoeia monographs 0452 and 0697. Currently this test is carried out in guinea pigs for each bulk of tetanus toxoid. To test the applicability of the in vitro BINACLE ("binding and cleavage") assay as an alternative method for the quality control of tetanus vaccines, two collaborative studies were run by the European Directorate for the Quality of Medicines & HealthCare under the aegis of the Biological Standardisation Programme. The first collaborative study indicated that the method allows sensitive TeNT detection. However, a clear conclusion could not be drawn due to the high variability of the results. To address the variability, the protocol was optimised and further standardised for the second study. The study results demonstrated good assay precision, both with respect to repeatability and reproducibility. Importantly, the limit of detection was 0.11 ng/mL TeNT in five out of nine laboratories and 0.33 ng/mL in four out of nine laboratories, suggesting that the BINACLE assay can detect TeNT with similar sensitivity as in vivo toxicity tests and can thus be taken into consideration as an alternative method to the current compendial in vivo test.

In 2010, the reporting of thrombotic adverse events for one subcutaneous and certain intravenous immunoglobulins (IGs) raised some concerns. In Europe, regulatory bodies rapidly revised compendial specifications for therapeutic IGs to ensure they do not exhibit thrombogenic (procoagulant) activity (PCA). At the global level, a working group (GWG) was launched with the aim of assessing PCA measurement methods and limits, considering results obtained by human IG manufacturers during in-process controls. The GWG created three dedicated subgroups to investigate the FXIa chromogenic assay, the non-activated partial thromboplastin time (NAPTT) test and the thrombin generation assay (TGA). The European Directorate for the Quality of Medicines & HealthCare (EDQM) was responsible for co-ordinating the subgroup in charge of evaluating the FXIa chromogenic assay in a study that assessed the sensitivity and robustness of two commercial chromogenic FXIa test kits. The impact of IG product formulation on FXIa recovery and the suitability of PCA-containing IG products as potential reference standards/controls were also assessed. IG materials representative of marketed products were provided to four laboratories for a study that was carried out in two steps: 1) two chromogenic FXIa test kit manufacturers assessed the performance and determined optimal test conditions by their respective methods, 2) two OMCLs studied both kits using an optimised study design. Regarding sensitivity, the study results identified suitable dose-response intervals and limits with both chromogenic FXIa test kits. This allowed the establishment of dilution ranges for optimal detection of FXIa/PCA in 5 % and 10 % IG products in the range of 1-6 mIU/mL. However, careful optimisation of the sample dilutions was required (notably to avoid potential matrix effects) and the choice of the mode of data acquisition (kinetic or end-point method) contributed to sensitivity in routine use. Importantly, the composition of IG products was of minor concern for FXIa determination with both test kits. Potential reference materials evaluated in the study behaved as expected and could be useful should a separate reference standard to the FXIa WHO IS be deemed necessary in future.

The level of anti-D antibodies in human immunoglobulin products for intravenous administration (IVIG) is controlled by the direct haemagglutination method prescribed by the European Pharmacopoeia (Ph. Eur.) that requires 2 control reference reagents. The World Health Organization (WHO) positive control International Reference Reagent (IRR; 02/228) with a nominal titre of 8 defines the highest acceptable titre, while the negative control preparation (02/226) has a nominal titre of <2. Working reference preparations (04/132 and 04/140) were subsequently established as Biological Reference Preparations (BRPs) for the Ph. Eur., and for distribution by the United States Food and Drug Administration (US FDA) and the National Institute for Biological Standards and Control (NIBSC). Due to diminishing stocks of these working reference preparations across the 3 institutions, a joint international study was organised to establish harmonised replacement batches. Sixteen laboratories contributed data to the study to evaluate positive and negative candidate replacement batches (13/148 and 12/300, respectively) against the WHO positive and negative control IRRs and the current working reference preparations (BRPs). The results show that the candidate reference preparations (13/148 and 12/300) are indistinguishable from the corresponding IRRs and current BRPs. The candidate preparations 13/148 and 12/300 were adopted by the Ph. Eur. Commission as Immunoglobulin (anti-D antibodies test) BRP batch 2 and Immunoglobulin (anti-D antibodies test negative control) BRP batch 2 with nominal haemagglutination titres of 8 and <2, respectively. The same materials were also adopted as NIBSC and US FDA reference preparations, thus ensuring full harmonisation.

For more than 50 years, in vivo assays have been used for testing pharmaceutical product safety due to their assumed ability to broadly detect potential unidentified contaminants. As part of these in vivo tests, the animal tests for depressor substances and histamine have been described in the European Pharmacopoeia since its first edition in 1977. Both tests measure the effect of histamine and histamine-like substances, using guinea-pigs and cats respectively. In 2024, the Histamine (2.6.10) general chapter is referenced in the Production section of four monographs and 10 monographs have variations of a sentence on designing the manufacturing process to eliminate or minimise substances lowering blood pressure in this same section, without referencing the chapter. The Depressor substances (2.6.11) chapter is referenced only in the Histamine (2.6.10) chapter as a next step if the histamine test is invalid. A historical search was performed and it has shown that the tests for histamine and for depressor substances were introduced by different groups of experts in an inconsistent way at different times, and for different reasons, leading to non-harmonised approaches across monographs. The control of histamine and other depressor substances has been the subject of numerous debates where their use was questioned. During these discussions, reports on positive cases or batches failing the test for histamine or depressor substances were anecdotal. In addition, in vivo tests can be considered non-specific, very variable, time-consuming, costly and ethically doubtful. More importantly, the majority of in vivo methods originate from a time when good manufacturing practice was not widely used and formal method validation requirements were not yet established. In view of the above, the removal of all references to animal tests for histamine or depressor substances from all texts still referring to them is proposed. Since the sentences in the Production section referring to the control of "substances lowering blood pressure", "vasoactive substances" or "hypotensive substances" appeared as remainders of the animal test without further guarantee of safety, it will also be proposed to remove all these sentences from the concerned monographs. Ultimately, the suppression of general chapters 2.6.10 and 2.6.11 from the Ph. Eur. is envisaged. Independently from the above, it is also envisaged to elaborate a new general chapter Histamine in active substances (2.5.47) to include physicochemical or immunochemical methods enabling the detection of histamine. This new text would aim at supporting manufacturers in their histamine control strategy following the inclusion of precaution statements in the general monograph on Products of fermentation (1468); these statements had been added in Ph. Eur. Supplements 9.6 and 10.4, following adverse events related to a GMP issue with gentamicin sulfate. This strategy has

The European Pharmacopoeia (Ph. Eur.) prescribes that hepatitis A vaccine (HAV) batches be tested for their potency before being released onto the European market. An enzyme-linked immunosorbent assay (ELISA) method for determination of antigen content of HAV was validated through an international collaborative study in 2012. This method requires specific coating and detection antibodies, i.e. coating reagent and detection antibodies Biological Reference Reagents (BRRs). The current batch of hepatitis A virus coating reagent (batch 1) was established in 2012. Stocks of this BRR were running low and a replacement batch was needed. Therefore, the European Directorate for the Quality of Medicines & HealthCare (EDQM) organised a collaborative study to qualify a replacement batch. Candidate coating reagent batch 2 was prepared from the same starting material as batch 1 and compared to the previous batch during the collaborative study. Results confirmed that candidate coating reagent batch 2 is suitable to be used for the intended purpose. The study demonstrated that batch 2 is comparable to batch 1 and that it can be used at the same working concentration, i.e. 1:500. The candidate material was adopted as Ph. Eur. Hepatitis A virus Coating Reagent for ELISA BRR batch 2 by the Ph. Eur. Commission at its 177th session in November 2023. It will be available from the EDQM under catalogue number Y0001624 once batch 1 is exhausted.

Human immunoglobulin products are used for the treatment of a number of diseases, such as primary or secondary immunodeficiencies and autoimmune conditions due to the complete absence of antibodies or the production of defective immunoglobulins. Quality control of human immunoglobulin products is essential to ensure therapeutic functionality and safety. This includes testing for Fc function and anticomplementary activity (ACA), as well as verification of appropriate molecular size distribution using size-exclusion chromatography as prescribed in the European Pharmacopoeia (Ph. Eur.) monographs 0338, 0918, 2788 and 1928. To this end, specific biological reference preparations (BRPs) must be used. Stocks of the Ph. Eur. Human immunoglobulin (molecular size) BRP were running low and therefore a collaborative study was run by the European Directorate for the Quality of Medicines & HealthCare (EDQM), under the aegis of the Biological Standardisation Programme, to calibrate replacement batches. Eighteen laboratories, including manufacturers and Official Medicines Control Laboratories, took part in the study. Three batches of candidate BRPs were assessed and compared to Ph. Eur. Human immunoglobulin (molecular size) BRP 3 to ensure continuity. Based on the study results, the candidate BRPs were adopted by the Ph. Eur. Commission as Ph. Eur. Human immunoglobulin (molecular size) BRP batch 4, 5 and 6.