Proposal (Title) Proposed 1St IS for Anti-Thyroglobulin Antibodies (Tgab)

WHO/BS/2021.2411 ENGLISH ONLY

EXPERT COMMITTEE ON BIOLOGICAL STANDARDIZATION Geneva, 18 to 22 October 2021

Requests to initiate new WHO reference material projects for biologicals

NOTE: This document has been prepared for the purpose of inviting comments and suggestions on the proposals contained therein, which will then be considered by the Expert Committee on Biological Standardization (ECBS). Comments MUST be received by 17 September 2021 and should be addressed to the World Health Organization, 1211 Geneva 27, Switzerland, attention: Technical Standards and Specifications (TSS). Comments may also be submitted electronically to the Responsible Officer: Dr Ivana Knezevic at email: [email protected].

This draft is intended for a restricted audience only, i.e. the individuals and organizations having received this draft. The draft may not be reviewed, abstracted, quoted, reproduced, transmitted, distributed, translated or adapted, in part or in whole, in any form or by any means outside these individuals and organizations (including the organizations' concerned staff and member organizations) without the permission of the World Health Organization. The draft should not be displayed on any website.

Please send any request for permission to:

Dr Ivana Knezevic, Technical Standards and Specifications, Department of Health Products Policy and Standards, World Health Organization, CH-1211 Geneva 27, Switzerland. Email: [email protected].

The designations employed and the presentation of the material in this draft do not imply the expression of any opinion whatsoever on the part of the World Health Organization concerning the legal status of any country, territory, city or area or of its authorities, or concerning the delimitation of its frontiers or boundaries. Dotted lines on maps represent approximate border lines for which there may not yet be full agreement.

The mention of specific companies or of certain manufacturers’ products does not imply that they are endorsed or recommended by the World Health Organization in preference to others of a similar nature that are not mentioned. Errors and omissions excepted, the names of proprietary products are distinguished by initial capital letters.

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This draft does not necessarily represent the decisions or the stated policy of the World Health Organization.

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Proposed new projects

1. WHO proposed 1st International Standard for anti-thyroglobulin

2. WHO proposed replacement for Factor IX concentrate

3 WHO proposed 5th International Standard for whole cell pertussis vaccine

4. WHO proposed 2nd International Standard for Interleukin-6

5. WHO Reference Panel for antibodies for SARS-CoV-2 Variants of concern

6. WHO 2nd International Standard for anti SARS-CoV-2 immunoglobulin

7. Use of whole-genome next generation (deep) sequencing for routine lot release of OPV and control of sIPV – developing test protocol and reference reagents

8. WHO Reference Reagent for CRM197

9. WHO 2nd IS for AFP, human cord serum

10. WHO proposed 5th IS for Hepatitis B Virus DNA for Nucleic Acid Amplification Techniques

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Proposal (title) Proposed 1st IS for anti-thyroglobulin antibodies (TgAb)

Proposer (name of NIBSC Principal contact Melanie Moore Institution)

Rationale Thyroglobulin autoantibodies (TgAb) are specific to the thyroglobulin protein (Tg) which plays a major role in thyroid hormone synthesis, storage and release. Autoantibodies to Tg are diagnostic of thyroid autoimmune diseases such as Hashimoto’s thyroiditis, Graves’ disease and other hyper/hypo thyroid diseases. Due to potential interference in Tg assay, TgAb may also be measured in tandem with Tg in the diagnosis and monitoring of differentiated thyroid cancers. Measurements of serum TgAb by immunoassay are an important component in the diagnosis of autoimmune thyroid disorders, and a WHO Reference Reagent (RR) for anti-thyroglobulin serum, coded 65/093, has been widely adopted for the calibration of these immunoassays. Stocks of the RR for anti-thyroglobulin serum, established by ECBS in 1970, are depleting and, at the current rate of dispatch (~95/year), will last until Q3 2023. As a result, there is now a requirement to prepare a replacement.

Anticipated uses and The standard is used by manufacturers as a calibrant for immunoassays for users anti-thyroglobulin autoantibodies; by clinical, regulatory and quality control laboratories and academic laboratories.

Source/type of materials The current RR 65/093 was produced from a pool of plasma samples from a number of donors containing antibodies to thyroglobulin. A similar approach will be used in the replacement, with plasma from 2-3 different donors with high titer TgAb, defibrinated and pooled to produce the bulk. Pooling donations from 2-3 donors is required to provide enough volume for ~2000 ampoules, and to help ensure coverage of the major epitopes.

Outline of proposed Major manufacturers of immunoassay kits and academic/clinical laboratories collaborative study (~10-12) will be invited to participate in an international collaborative study with the following aims: 1 – to confirm reactivity of the 1st IS by immunoassay, and assess the relationship of the activity of the 1st IS with the RR 65/093 and existing local standards 2 – to calibrate the candidate 1st IS for anti-TgAb against the RR 65/093 3 – to assess stability of ATD samples of the 1st IS by immunoassay 4 – to assess the commutability of the candidate 1st IS with the inclusion of serum and/or plasma samples Participants will be asked to report data to NIBSC for statistical analysis and a consensus will be reached on a unitage.

Issues raised by the None proposal

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Action required ECBS to endorse proposal

Proposer's project tbc Date proposed: October 2021 reference

CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES (TRS932)

Approval status of Diagnostic. Defines the IU for current TgAb immunoassays. medicine or in vitro diagnostic method

Number of products or ~20 major manufacturers of immunoassays for TgAb, majority of whom are methods traceable to the RR 65/093.

Public health Serum TgAb measurements, reporting in IU, are used on a worldwide basis importance to diagnose autoimmune thyroid diseases. The current RR, 65/093, is used for the calibration of these immunoassays.

Global importance Thyroid auto-immune disease is of global importance. The current RR 65/093 is distributed worldwide. Immunoassays for measurement of TgAb

are manufactured and marketed worldwide.

Global need from There is a need for NIBSC to provide continuity of the International Unit for regulatory & scientific TgAb through the production of an International Standard. considerations

ECBS outcome [BLANK]

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Proposal (title) The 6th International Standard for Blood Coagulation Factor IX Concentrate

Proposer (name of NIBSC Principal contact Stella Williams Institution)

Rationale The 5th International Standard (IS) for FIX is used for potency labelling of all Factor IX (FIX) and Prothrombin Complex Concentrate (PCC) therapeutics. Stocks of the 5th International Standard for Blood Coagulation Factor IX as well as the 3rd BRP for Human Coagulation Factor IX concentrate will be depleted in the next couple of years and will therefore need to be replaced. The current IS and Ph Eur BRP originate from the same batch of ampoules, supporting the traceability of the IU. EDQM has expressed a wish to maintain this harmonized approach and proposed to share the same batch of material for the 6th IS and 4th BRP. Since the establishment of the 4th IS in 2008, a number of modified and extended half-life (EHL) FIX products have been licensed, with potency value assigned relative to the 4th IS, with the labelled IU supported by clinical trial data. However, substantial assay discrepancies exist for EHL products when assayed using different reagents and kits relative to the IS. To mitigate the risk of discontinuity of the IU related to these products, the same plasma derived FIX preparation as the 4th IS was established as the 5th IS. A similar approach is now proposed for the 6th IS and the same plasma derived FIX product has been sourced as a candidate. Approximately 20,000 ampoules of 5th IS and 3rd BRP were produced in 2015 and replacements are needed within 6/7 years of establishment, so a larger batch would be desirable. In addition, the plasma derived FIX product, used as bulk for the 4th and 5th IS, is being phased out by the manufacturer and there may not be any of this product available when the replacement of the 6th IS would be required in 5 to 6-years’ time. Therefore, enough candidate material has been donated for NIBSC to fill 2 or 3 x 25,000 ampoules of candidate preparations and this will ensure that we will have sufficient material for the next 12 - 18 years. Candidates from all fills will be included in the same collaborative study.

Anticipated uses Approximately 600 ampoules of the IS for FIX are dispatched each and users year with the majority being procured by FIX product manufacturers, regulators and national control laboratories and used for potency labelling and batch release testing of FIX therapeutics.

Source/type of The candidate material will be a plasma-derived concentrate FIX, materials

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similar or the same source as the 5th International standard to try to

ensure continuity of the unit and consistency for all FIX products including the modified and EHL products that show discrepancies between assay methods and reagents.

Outline of proposed Due to the need to assess the impact of the candidate preparation collaborative study on EHL products the collaborative study will be carried out in two phases: Phase 1 - An in-house (NIBSC) investigation with the support of FIX product manufacturers: A selection of currently licensed plasma derived, recombinant and EHL products will be assayed against the candidate material alongside the 5th IS for FIX using multiple methods and reagents to ensure the continuity of the IU for all products. Phase 2 - Value Assignment: the candidate will be assayed against the current (5th) IS in an international collaborative study involving multiple participants, using their routine clotting and chromogenic methods.

Issues raised by the None proposal

Action required ECBS to endorse proposal

Proposer's project BIO00144 Date proposed: reference

CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES (TRS932)

Approval status of >20 different plasma derived, recombinant and extended half-life medicine or in vitro FIX therapeutics are licensed around the world and with more in diagnostic method development.

Number of products There are 2 methods for measuring the potency of these products: or methods One-stage clotting and Chromogenic substrate assays. Within these 2 methods there are a number of different reagents/kits.

Public health FIX therapeutics are on the WHO list of essential medicines and are importance used to treat haemophilia B: a congenital deficiency in factor IX that results in uncontrolled bleeding. Prothrombin Complex Concentrates which contain FIX are used for reversal/treatment of Warfarin overdose.

Global importance Accurate potency labelling of FIX products allows for consistent and harmonized dosing of haemophilia B patients around the world.

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Global need from The International Standard provides the continuity of the IU to regulatory & ensure the safety and efficacy of FIX products. scientific considerations

ECBS outcome [BLANK]

Running Title: FIX Concentrate (6th IS)

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Proposal (title) Proposed fifth International Standard for whole cell pertussis vaccine

Proposer (name of NIBSC Principal contact Alex Douglas-Bardsley Institution)

Rationale The 4th International Standard for Bordetella pertussis vaccine (whole cell) is a freeze dried suspension of killed B. pertussis donated by CSL Australia in 1994. It was established in 2005 with a unitage of 40 IU/ampoule. The current preparation has the code number 94/532. The stock of the 4th IS is running low and a replacement is needed. It is proposed to replace this material in the next two years to ensure a continuous supply.

Anticipated uses and The potency of whole cell pertussis vaccines is measured in IU based users on the IS. Therefore, it is widely used to calibrate in-house reference preparations in the potency control test (Kendrick test) for whole cell pertussis vaccines. The intended users of the product are vaccine manufacturers and control laboratories Approximately 150-200 ampoules are distributed per year. However, this product is on restricted sale and therefore the actual demand is more.

Source/type of The Serum Institute India have agreed to donate whole cell pertussis materials material to produce this standard. The proposed batch size is 4,000 – 5,000 ampoules which at the current rate of sale, without current restrictions, will allow for 20 years stock.

Outline of proposed Value will be assigned using the in vivo Kendrick test relative to the collaborative study 4th IS. It is proposed to involve approximately 10-15 laboratories in the collaborative study. The laboratories will be made up of NCLs and vaccine manufacturers.

Issues raised by the While the manufacturer has kindly offered to donate the material, the proposal pandemic has caused a delay in sourcing the material from them.

Action required ECBS to endorse proposal

Proposer's project Date proposed: 2021 reference

CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES (TRS932)

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Approval status of Whole cell vaccines are the widely used whooping cough vaccines medicine or in vitro and there are many licensed vaccines from a large number of diagnostic method manufacturers available worldwide.

Number of products Being widely used globally in medium and low income countries, there or methods are several licensed whole cell pertussis containing vaccines available. The standard is to be used by laboratories to calibrate their own reference preparations for use in the potency assay (Kendrick test).

Public health Whole cell pertussis containing vaccines remain part of routine importance vaccination programmes in a number of countries across the world.

Global importance Pertussis infection is still the cause of significant mortality and morbidity worldwide. Whole cell pertussis vaccines are used globally.

Global need from An international standard is important for vaccine manufacturers and regulatory & scientific NCLs to calibrate their reference preparations ensuring accurate considerations evaluation of whole cell vaccines.

ECBS outcome

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Proposal (title) Proposed 2nd International Standard for Human Interleukin-6 (IL-6)

Proposer (name of NIBSC Principal contact Meenu Wadhwa Institution)

Rationale Increased usage of the 1st WHO IS for IL-6 in recent years including the COVID-19 pandemic is resulting in rapid depletion of the current stock of the 1st IS, established by ECBS in 1992. At the current rate of dispatch, it is anticipated that the stocks will last until Q3/4 2022. Consequently, there is now an urgent requirement to prepare a replacement IS. IL-6 is a pleiotropic cytokine associated with regulation of inflammation, Anticipated uses and hematopoiesis, cancer progression and immune responses. As a result, the users current 1st WHO IS (coded 89/548; 100, 000 IU/ampoule) for IL-6 is primarily used by manufacturers (biotherapeutics, immunoassays) and researchers for calibrating IL-6 preparations with use in a broad range of applications. This includes its role as a critical reagent in cell-based assays for potency testing of monoclonal antibodies targeting IL-6 (e.g., siltuximab, Sylvant ®) and IL-6 receptor (e.g., tocilizumab, Actemra/RoActemra®; sarilumab, Kevzara®) which are approved for various indications. The IS is also used for potency testing of IL-6 preparations for use as cell culture supplement for cell and gene therapy products (ex vivo maturation of immature dendritic cells and expansion of haemopoietic stem cells). Additionally, it has significant use in immunoassays for measuring IL-6 levels in (a) tissues as biomarker of inflammation or disease pathology in clinical settings e.g., sepsis, autoimmune, infectious diseases etc, (b) in pre-clinical testing and/or clinical testing of immunotherapies (e.g., cytokine release syndrome associated with mAbs, CAR-T cells) and (c) in diverse research programs. Source/type of materials Recombinant human IL-6 to be sourced from commercial supplier.

Outline of proposed Recombinant human IL-6 will be lyophilized and a collaborative study which collaborative study includes both bioassays and immunoassays will be organised to evaluate the suitability of the candidate lyophilized preparation as WHO IS. Calibration will be carried out relative to the 1st WHO IS for IL-6. Participant data will be statistically analysed and a consensus will be reached on the unitage.

Issues raised by the None proposal

Action required ECBS to endorse proposal

Proposer's project Date proposed: 5th May 21 reference

CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES (TRS932)

Approval status of Critical reagent for potency evaluation of licensed anti-IL-6 (e.g., Sylvant ®) medicine or in vitro and anti-IL-6 receptor antibodies (e.g., Actemra/ RoActemra ®, Kevzara®). diagnostic method

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Number of products or At least 3 approved antagonists; others are in clinical trials (e.g., methods clazakizumab, olokizumab – both targeting IL-6). Currently, several biosimilars of tocilizumab are in development.

Public health Replacement IS is important for controlling the bioassay performance for importance consistent potency evaluation of anti-IL-6/anti-IL-6 receptor products and for calibrating immunoassays that are in use for measuring IL-6 levels.

Global importance Helpful for consistent manufacturing of cell and gene therapy products, for potency of the approved anti-IL-6/anti-IL-6 receptor products particularly as

many tocilizumab biosimilars are in development. Promote global harmonisation of bioassays and immunoassays.

Global need from The availability of IL-6 IS would help in continued calibration of IL-6 regulatory & scientific preparations used for potency of IL-6/IL-6 receptor antagonists and global considerations harmonisation of assays used for clinical testing and immunotherapy programs.

ECBS outcome [BLANK]

Running Title: 2nd IS for IL-6

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Proposal (title) WHO Reference Panel for antibodies for SARS-CoV-2 Variants of Concern

Proposer (name of NIBSC Principal contact Giada Mattiuzzo Institution)

Rationale Severe acute respiratory syndrome coronavirus -2 (SARS-CoV-2) is the aetiological agent of the 2020-21 pandemic of Coronavirus Disease 2019 (COVID-19). On 10th December 2020 WHO Expert committee on Biological Standardization established the 1st WHO International Standard for anti-SARS-CoV-2 immunoglobulin and WHO International Reference Panel. However, towards the end of 2020, SARS-CoV-2 variants with mutations which render them resistant to therapeutic monoclonal antibodies and convalescent plasma from COVID-19 patient from early 2020 started to emerge. These new isolates defined Variant of Concern (VOC) may have a critical impact on vaccines and therapeutics efficacies and reinfection. A Reference Panel constitute of convalescent plasma or serum from recovered COVID19 patients infected with one of the VOCs (as of June 2021: https://www.who.int/en/activities/tracking-SARS-CoV-2- variants/) will facilitate the development of serological assays to study the effect of the VOC.

Anticipated uses and Standardisation of serological assay (e.g. ELISA, neutralisation users assays) for identification and/or potency test of anti-SARS-CoV-2 antibodies in: Clinical and public health laboratories Vaccine manufacturers - Vaccine studies Therapeutics strategies (mAbs, convalescent plasma) Assay Kit manufacturers Research laboratories

Source/type of materials Donated human serum or plasma from convalescent individuals who have been infected with one of the VOC, currently alpha/B.1.1.7, beta/B.1.351, gamma/P.1 and delta/B.1.617.2. The project is supported by the Coalition for Epidemic Preparedness Innovations (CEPI). Several organizations have answered CEPI/NIBSC and WHO call to source such material and we are currently selecting collaborating partners for these donations. Materials issued by NIBSC will have undergone treatment steps for virus inactivation, to be shipped as non-infectious. All clinical and other samples will have undergone screening for blood-borne viruses.

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Outline of proposed Collaborative study will involve 15-20 laboratories worldwide, performing a collaborative study range of serological assays for SARS-CoV-2 VOC. In the same study, the replacement for the WHO International Standard for anti-SARS-CoV-2 immunoglobulin will be also evaluated. Laboratories will be selected based on ability to perform several assays to compare the effect of VOCs. The aims will be to assess the suitability of different antibody preparations to serve as reference material for the serological assay for SARS-CoV-2 VOC • characterisation of the antibody preparations in terms of reactivity/specificity in different assay systems. • assessing each preparation’s potency i.e. readout in a range of typical assays performed in different laboratories

Issues raised by the The study is taking place during COVID-19 pandemic, and logistics could be proposal difficult. Personnel may also be an issue due to mobility restrictions and self- isolation.

Action required ECBS to endorse proposal

Proposer's project Date proposed: 15 June 2021 reference

CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES (TRS932)

Approval status of As 25th May 2021 there are 101 vaccines at clinical stage and 8 vaccines medicine or in vitro approved for emergency use in at least one country worldwide. There are diagnostic method over 450 clinical trials evaluating therapeutics, such as monoclonal antibodies, anti-viral, immunomodulators, cell and gene therapy. Nine have received authorization for emergency use.

Number of products or There are at least 77 assays who have received emergency use methods authorization and comprises of ELISA, surrogate neutralisation, rapid test, immunofluorescence-based methods. These assays target a wide variety of viral antigens (RBD, S, S1, N) and different class of immunoglobulins.

Public health The World Health Organization declared COVID-19 a Public Health importance Emergency of International Concern on 30th January 2020, and a th Pandemic on 11th March 2020. As 25 May 2021, approx. 170 million of people have been infected and 3.5 million deaths have been recorded.

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Serological assays against VOCs are therefore of utmost importance.

Global need from Standardised and calibrated assays are vital for accurate evaluation of regulatory & scientific treatments, including antibody therapies and vaccine, and for case considerations management and surveillance.

ECBS outcome [BLANK]

Running Title: SARS-CoV-2 VOC reference panel

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Proposal (title) 2nd WHO International Standard for anti-SARS-CoV-2 immunoglobulin

Proposer (name of NIBSC Principal contact Giada Mattiuzzo Institution)

Rationale Severe acute respiratory syndrome coronavirus -2 (SARS-CoV-2) is the aetiological agent of the 2020-21 pandemic of Coronavirus Disease 2019 (COVID-19). On 10th December 2020 WHO Expert committee on Biological Standardization established the 1st WHO International Standard (IS) for anti-SARS-CoV-2 immunoglobulin and WHO International Reference Panel. Approximately 3000 ampoules were available for distribution. As 1st May 2021, over 1800 ampoules were distributed to 468 end users worldwide. This is despite a restriction of 5 ampoule per end user. At the current rate, the IS will be depleted within one year. Also, the suitability of the IS against SARS-CoV-2 variant of concern (VOC) requires continuous monitoring. Currently, the 4 VOCs are all neutralized at some degree by the IS, making the standard still fit for purpose. Nevertheless, to ensure continuity of the IS unitage a replacement is needed.

Source/type of materials Donated human serum or plasma from convalescent individuals who have been infected with one of the VOC, currently B.1.1.7, B.1.351, P.1, B.1.617. Convalescent plasma with high antibody titer against all the variant will be the preferred candidate material. The project is supported by the Coalition for Epidemic Preparedness Innovations (CEPI). Several organizations have answered CEPI/NIBSC and WHO call to source such material and we are currently selecting collaborating partners for these donations. Materials issued by NIBSC will have undergone treatment steps for virus inactivation, to be shipped as non-infectious. All clinical and other samples will have undergone screening for blood-borne viruses.

Outline of proposed The candidate replacement IS will be evaluated in parallel with the proposed collaborative study panel of convalescent plasma/serum from individual recovered from SARS- CoV-2 VOC.

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Collaborative study will involve 15-20 laboratories worldwide, performing a range of serological assays for SARS-CoV-2 VOC. Laboratories will be selected based on ability to perform several assays to compare the effect of VOCs. The aims will be to assess the suitability of candidate replacement to serve as reference material for the serological assay for SARS-CoV-2 • characterisation of the antibody preparations in terms of reactivity/specificity in different assay systems; this will include also performance against different VOCs • assessing each preparation’s potency i.e. readout in a range of typical assays performed in different laboratories • Evaluate performance of the replacement IS in parallel to the current First WHO IS for SARS-CoV-2 immunoglobulin and propose an appropriate unitage to assure continuity between the first and second IS • ability to harmonise assays results between SARS-CoV-2 serological assay at least between the same isolate, but ideally between different VOCs

Issues raised by the The study is taking place during COVID-19 pandemic, and logistics could be proposal difficult. Personnel may also be an issue due to mobility restrictions and self- isolation.

Action required ECBS to endorse proposal

Proposer's project Date proposed: 15 June 2021 reference

CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES (TRS932)

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Global importance The impact of COVID-19 pandemic is not only limited to the death rate associated. Economical impact due to lockdown measurements, restriction in travel moment are difficult to estimate. Vaccination programmes are seen as the exit strategy in many countries. Maintain the availability of the WHO International Standard for anti-SARS-CoV- 2 immunoglobulin to facilitate comparability in the analysis of humoral responses and evaluation of a possible correlate of protection. It is also important to evaluate the efficacy of the vaccines against VOCs. Serological assays against VOCs are therefore of utmost importance

ECBS outcome [BLANK]

Running Title: Replacement SARS-CoV-2 antibody standard

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Proposal (title) WHO Reference Reagent for CRM197

Proposer (name of NIBSC Principal contact Barbara Bolgiano / Paul Stickings Institution)

Rationale “Cross-reacting material 197” or CRM197 is a non-toxic mutant of diphtheria toxin that is used as a carrier protein in a number of different polysaccharide conjugate vaccines, including Meningococcal, Pneumococcal and Typhoid glycoconjugates. CRM197 can be produced using a “traditional” C. diphtheriae expression system although the relatively low yields of this approach have led to a number of companies to explore production of recombinant CRM197 using heterologous expression systems. For use in vaccine manufacturing CRM197 is characterized using immunochemical and physicochemical methods. The availability of a WHO reference reagent will provide a source of stable, well-characterized material that can serve as a control for some of the analytical methods that are applied during QC.

Anticipated uses and The reference reagent is intended for use as a control material in users immunochemical and physicochemical methods used in QC

Source/type of materials An adequate source of E. coli-derived material has been offered for this purpose.

Outline of proposed A collaborative study will be run to demonstrate fitness for purpose but it is collaborative study not anticipated that units of activity will be assigned to this material. The design of the collaborative study will reflect this. The carrier protein content is quoted in SI units, normally in µg/ml in bulk conjugates and µg/dose (or range of ug/dose) in final product. Protein content can be obtained most accurately using UV spectrophotometry which would be a primary method, as the extinction (absorption) coefficients are known and based on a physical parameter, namely the absorption of UV light of 280 nm by the aromatic amino acids of the protein.

Issues raised by the It will be important to establish if a rCRM197 can be an adequate control for proposal native CRM197 and Pseudomonas fluorescens CRM197

Action required ECBS to endorse proposal

Proposer's project TBC Date proposed: July-2021 reference

CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES (TRS932)

Approval status of The following conjugate vaccines containing CRM197 have been pre- medicine or in vitro qualified: MenACWY-CRM197 (native), Pneumococcal 10-valent-CRM197 diagnostic method vaccines (native and P. fluorescens) and Vi-CRM197 (native). Additional CRM197-conjugated vaccines are expected to be submitted for prequalification including a Group B streptococcal CRM197 (native) vaccine.

Number of products or A CRM197 reference material could serve as a control in the following

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methods types of assays: identity, purity, nicking or integrity and free protein. The methods could include latex agglutination, ELISA, electrophoresis (SDS- PAGE, reducing or non-reducing, Western blotting) HPLC size-exclusion chromatography and capillary zone electrophoresis.

Public health CRM197 has provided an important alternative to tetanus toxoid and other importance carrier proteins, and is expected, as it has not contributed to the supression of the immune response to other antigens. It is expected to remain a carrier

protein of choice in the future.

Global importance CRM197 is being produced by manufacturers in SEARO and is being used in vaccines for SEARO and AFRO regions.

Global need from Although CRM197 has not been evaluated for its protection against regulatory & scientific diphtheriae or licensed as a substitute for diphtheria toxoid in its own right, considerations its quality, consistency of production (and conjugation), immunogenicity and stability are important for the protection given by conjugate vaccines. There is no difference in the requirements for CRM197 in different regions.

ECBS outcome [BLANK]

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Proposal (title) Use of whole-genome next generation (deep) sequencing for routine lot release of OPV and control of sIPV - developing test protocol and reference reagents

Proposer (name of CBER - NIBSC Principal contact Kostya Chumakov - Javier Martin Institution)

Rationale Stability of genetic characteristics of viral vaccines is an important aspect of their safety. Sabin strains used in manufacture of OPV accumulate reversions during replication in humans and cell cultures that may increase virulence. Traditionally, genetic stability of OPV is tested in vivo by measuring changes in the attenuation phenotype using monkeys or transgenic mice expressing the human poliovirus receptor neurovirulence tests (NVTs) and in vitro by measuring the proportion of revertant mutations using MAPREC. Two years ago, a first phase of an ECBS-endorsed International Collaborative Study jointly coordinated by NIBSC and CBER/FDA showed that next generation sequencing (NGS) could be used as a replacement of MAPREC for measuring the content of neurovirulent mutants in the 5’-UTR region. In addition, NGS could be used to determine a profile of single-nucleotide polymorphisms (SNP) in the entire genome of vaccine poliovirus. Comparison of the SNP profiles could be used for monitoring consistency of vaccine production. NGS analysis can be a very sensitive tool for quality control of OPV ensuring that no reversions take place anywhere in the viral genome during manufacture. NGS can also be used to ensure that attenuated characteristics of the virus are retained in the course of Sabin IPV (sIPV) manufacture. Therefore, a combination of quantitative analysis of neurovirulent mutations in the 5’-UTR (MAPREC replacement) and comparison of whole-genome SNP profiles might be used for routine lot release of OPV as a replacement of in vivo neurovirulence testing, leaving monkey and transgenic mouse NVTs for the qualification of seed viruses during the establishment phase. The purpose of this project is to develop a testing procedure/protocol including criteria for comparing SNP profiles to make decisions about consistency of manufacture, and to create reference reagents suitable for NGS-based quality control of OPV and sIPV.

Anticipated uses and The study will result in the development of a method and reference reagents users for monitoring consistency of OPV and sIPV manufacture that could be used for routine lot release of these products as a replacement for animal based NVTs. They will be used by vaccine manufacturers and National Control Laboratories.

Source/type of materials The reference materials will be made from Sabin strains grown in cell culture

Outline of proposed The study will be coordinated by NIBSC and CBER/FDA using materials collaborative study provided by OPV and sIPV manufacturers. Manufacturers and National Control Laboratories from worldwide will participate in the study. Participating labs will be given an opportunity to either generate SNP profiles in house, provide materials to NIBSC and FDA, or use contract research laboratories. Bioinformatic analysis of the results will be performed by NIBSC and CBER/FDA.

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Issues raised by the None proposal

Action required ECBS to endorse proposal

Proposer's project Date proposed: May 2021 reference

CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES

Approval status of A number of OPV and sIPV containing vaccine products are approved in medicine or in vitro many countries, with a significant number of additional manufacturers diagnostic method expected to start sIPV production in the near future. A molecular method for OPV lot release (MAPREC) is included in "Recommendations to assure the quality, safety and efficacy of poliomyelitis vaccines (oral, live, attenuated)", TRS980. The document also mentions the potential use of NGS but does not specify the protocol and reference materials needed. The proposed study of NGS utility as a replacement of in vivo neurovirulence test for routine lot release could close this gap. ECBS meeting in 2020 proposed to revise TRS980, therefore the results of this study could inform the revision.

Number of products or OPV of all three serotypes, Sabin IPV, and novel OPV (nOPV) made from methods genetically stabilized strains. The study aims at the replacement of MAPREC and in vivo NVTs for routine lot release of these vaccines.

Public health Polio eradication still requires mass quantities of OPV, and in the future, importance nOPV. After polio eradication the use of IPV (including sIPV) will be greatly expanded. Introduction of NGS for quality control of all these products will

be a significant relief for vaccine manufacturers and NCLs and improve both vaccines quality and their supply. Besides, animal testing by NVT which uses the largest number of animals for any in vivo test will be removed.

Global importance Polio eradication remains one of the top global priorities of WHO. Even after the eradication there will be a need for continuous supply of safe and

effective polio vaccines. The proposed approach will improve quality and simplify their manufacture and control.

Global need from The proposed approach is based on the cutting-edge science and regulatory & scientific technology and may represent the first example of a molecular-genetic considerations method based on full-genome deep sequencing used for quality control and lot release of a vaccine. If successful, it may set a new paradigm for vaccine quality control that may be used for other high profile viral vaccines such as yellow fever vaccines.

ECBS outcome [BLANK]

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Proposal (title) The 2nd WHO IS for AFP, human cord serum

Proposer (name of NIBSC Principal contact Katherine Partridge Institution)

Rationale Stocks of the 1st IS for AFP, human, cord serum-derived (72/225) are depleting and, at the current rate of dispatch, will last until 2023. As a result, there is now a requirement to prepare a replacement.

Anticipated uses and The users will be manufacturers of AFP diagnostic tests and related users materials, such as antigens. The material is predominantly used to calibrate test kits measuring AFP levels in patients’ urine/serum/amniotic fluid in International Units.

Source/type of materials The 1st IS consisted of pooled human cord serum. For the replacement IS we will aim to procure a similar appropriate purified material from an external supplier.

Outline of proposed Participants in the collaborative study are likely to be manufacturers or collaborative study clinical laboratories who perform AFP assays. They will be asked to calibrate the candidate preparation in terms of the 1st IS by AFP immunoassays. They will be asked to report data to NIBSC for statistical analysis and a consensus will be reached on a unitage.

Issues raised by the None proposal

Action required ECBS to endorse proposal

Proposer's project TBC Date proposed: 09/07/2021 reference

CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES (TRS932)

Approval status of Human AFP is used in many in vitro diagnostic methods in the diagnosis of medicine or in vitro some cancers, liver disease, and congenital abnormalities. diagnostic method

Number of products or >10 products and methods methods

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Public health AFP products and diagnostic kits, dosed/calibrated in International Units in importance terms of the International Standard, are used on a worldwide basis. Inaccurate dosing can result in inaccurate diagnoses of numerous disease

states/conditions. AFP is used in the diagnosis of some cancers, including hepatocellular carcinoma (HCC). Levels are also monitored through the duration of HCC treatment as an indicator for responsiveness. AFP levels may also be measured in the diagnosis of liver disease, as well as “triple” and “quadruple” pregnancy screening tests, looking for congenital abnormalities including neural tube defects and gastroschisis.

Global importance Recombinant and native AFP products are manufactured and marketed worldwide. In order to maintain quality of care and availability of these

diagnostic tests, there is a continued need for effective standardization.

Global need from There is a need for NIBSC to provide continuity of the International Unit of regulatory & scientific AFP immunoreactivity through the continued provision of an International considerations Standard.

ECBS outcome [BLANK]

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Proposal (title) Proposed 5th WHO International Standard for Hepatitis B Virus DNA for Nucleic Acid Amplification Techniques

Proposer (name of NIBSC Principal contact Jacqueline Fryer Institution)

Rationale The WHO International Standard for HBV DNA is used in the calibration of secondary reference materials and in the validation of HBV NAT assays used in the safety testing of blood donations, and cells, tissues and organs. HBV NAT is also widely used in the management of HBV infection, particularly in the diagnosis of disease and the initiation and monitoring of antiviral therapies. As of Jul 2021, the existing stock of the current 4th HBV IS (10/266) is ~600 vials. At current rates of issue stocks are likely to be exhausted in 2 years. The replacement will require up to 2 years for completion.

Anticipated uses and Used to calibrate secondary reference materials and in the validation of HBV users NAT assays. Anticipated users are; blood transfusion centres, clinical laboratories, control authorities, and IVD manufacturers.

Source/type of materials The 4th HBV IS (10/264) is a dilution of the Eurohep reference material, R1 (genotype A, HBsAg subtype adw2), in pooled human plasma. There is sufficient residual stock of this Eurohep R1 reference to make ~3000-3500 vials (0.5 mL volume) of candidate.

Outline of proposed Study (comprising 10-20 participants) to calibrate new candidate against collaborative study 10/266.

Issues raised by the None proposal

Action required ECBS to endorse proposal

Proposer's project IDD00019 Date proposed: October 2021 reference

CONSIDERATIONS FOR ASSIGNMENT OF PRIORITIES (TRS932)

Approval status of NAT is the recommended method for the detection of HBV DNA in human medicine or in vitro serum and plasma. diagnostic method

Number of products or A range of NAT-based methods are available for the detection and methods quantification of HBV DNA, including both commercial and laboratory- developed assays.

Public health The need to standardize NAT-based assays for HBV is ongoing. NAT is importance routinely used to manage HBV infections and there remains a major risk of transfusion-transmitted infection due window-period donations, vaccine

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breakthrough infections and occult HBV infection.

Global importance HBV remains a major global health problem. WHO estimates that in 2015, 257 million people were living with chronic hepatitis B infection (defined as

hepatitis B surface antigen-positive). In 2015, hepatitis B resulted in an estimated 887 000 deaths, mostly from cirrhosis and hepatocellular carcinoma (i.e. primary liver cancer).

Global need from A number of countries are mandated to screen cells, tissues and organs for regulatory & scientific transplantation for HBV by NAT. Although blood screening for HBV by NAT considerations is not yet not mandated, it has been widely implemented on a voluntary basis.

ECBS outcome [BLANK]

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