90th Meeting – August 16, 2007

Bethesda, MD


Informational Presentations:  FDA/WHO Biological Standards



A.  Summary of January 29-30, 2007 WHO Meeting with WHO Collaborating Centres for Biological Standards and Standardization to Support the Development of WHO Biological Reference Preparations for Blood Safety-related in vitro Diagnostic Tests, Paul A. Mied, Ph.D., DETTD, OBRR, FDA


WHO Collaborating Centres for Biological Standards and Standardization:

  • National Institute for Biological Standards and Control (NIBSC)
  • Paul-Ehrlich-Institut (PEI)
  • Center for Biologics Evaluation and Research (CBER)


Meeting Convened by:

The Quality Assurance and Safety: Blood Products and Related Biologicals (QSD) team, Department of Medicines Policy and Standards (PSM), World Health Organization (WHO)


Location of the Meeting:

FDA/CBER, Bethesda, Maryland


Objectives of the Meeting:

  • To foster cooperation among the WHO Collaborating Centres for Biological Standards and Standardization.
  • To strengthen the development of WHO International Biological Reference Preparations for the control of in vitro diagnostic tests related to blood safety.


Intended Use of the WHO Biological Reference Preparations:

Validation, quality control, assessment of comparability, and regulation on a global basis of blood safety-related in vitro diagnostic tests, contributing to a harmonized regulation of blood and blood products. 


Priority Projects Identified:    



  1.  Replacement of Existing WHO Biological Reference Preparations near   exhaustion:



1.  Anti-HBs immunoglobulin (proposed 2nd International Reference



2.  HCV RNA (proposed 3rd International Standard)


3.  Parvovirus B19 DNA (proposed 2nd International Standard)


4.  Anti-Syphilitic Standard (proposed 2nd International Standard)



  1.  New WHO Biological Reference Preparations Needed:



5.  HIV-1 Genotype Panel (proposed  2nd International Reference Panel)


6.  HIV-2 RNA International Standard


7.  Anti-HIV Panel (proposed 2nd International Reference Panel)


8.  HBsAg and HBV DNA Genotype Panels


9.  Anti-HBc International Standard


10.  Anti-HCV Panel


11.  Anti-HTLV-I/II Panel


12.  Anti-Plasmodium species Panel


13.  Anti-Trypanosoma cruzi  Preparation/Panel


C.  Proposed WHO Biological Reference Preparations that need further discussion: 



14.  HIV-2 RNA Genotype Panel


15.  HCV Genotype Panel


16.  Parvovirus B19 Genotype Panel


17.  Anti-CMV Standard


18.  WNV RNA Preparation/Pan Panel for arthropod-borne flaviviruses RNA


19.  HCV Core Antigen Preparation


20.  Anti-HHV-8 and HHV-8 DNA Preparations


21.  TSE Blood Preparations


22.  Blood-borne Bacteria Panel


23.  Anti-Leishmania Panel


Additional Agreements among WHO Collaborating Centres:


·     A need for collection and exchange of epidemiological information with an impact on blood safety. 


·     The established WHO Biological Reference Preparations and those to be developed are suitable to cover the forthcoming new technologies for detection of infectious agents.


·         A need for improved collaboration among WHO Collaborating Centres and between the WHO Collaborating Centres and WHO.  Annual face-to-face meetings and teleconferences are necessary to monitor progress. 


·         A need to establish a network of WHO Collaborating Centres for in vitro diagnostics-related biological standardization representing all WHO Regions to ensure complementary and focused expertise at the global level.



Plan of Action:

The Priority Projects to establish WHO Biological Reference Preparations to support international regulations for blood and blood products safety will form a five-year In Vitro Diagnostic strategic plan.  This plan will be submitted to the ECBS for endorsement in October, 2007.  The In Vitro Diagnostic strategic plan will be used to support resource mobilization activities.

B.        Potency and Safety Standards for Plasma Derivatives, Mei-ying W. Yu,           Ph.D., DH, OBRR, FDA


The Division of Hematology (DH) in CBER/FDA has the primary responsibility for the scientific evaluation of manufactured biologic products derived from blood or plasma and their analogs derived from recombinant DNA technology.  Examples of these products include clotting factors, immune globulins, and albumin.   To ensure their safety and effectiveness, DH personnel have actively participated in developing and establishing CBER, WHO or global potency and safety standards through close collaboration with WHO collaborative centers (e.g., NIBSC and PEI), EDQM (Ph Eur), and industry.  CBER standards are available upon request to product manufacturers for lot release testing of final-container products or for in-process control testing.  


Potency Standards for Lot Release Testing for Clotting Factors: 





Potency per vial or ampoule


Ref. #

FIX Complex,  FIX, rFIX

WHO 3rd IS for FIX Concentrate (NIBSC 96/854)/Ph Eur/CBER

  10.7 IU FIX 



CBER Mega 2/Ph Eur  Batch 3 Standard for FVIII  

10.4 IU FVIII (clotting assay)

8.6 IU FVIII (chromogenic assay) 



WHO 1st IS for vWF Concentrate (NIBSC 00/514)

9.4 IU vWF:Ristocetin Cofactor



Thrombin in

Fibrin Sealant,   bovine thrombin


WHO 2nd IS for Thrombin (NIBSC 01/580)/CBER Lot K

110 IU Thrombin



Abbreviations:  IS, International Standard; FIX Complex, FIX Complex (Human); FIX, Coagulation Factor IX (Human); rFIX, Coagulation Factor IX (Recombinant); AHF, Antihemophilic Factor (Human); rAHF, Antihemophilic Factor (Recombinant); AHF/vWF, Antihemophilic Factor/von Willebrand Factor Complex (Human)  


Potency and Safety Standards for Immune Globulins or Albumin:





Potency Requirement or Potency per mL (or when reconstituted)

Ref. #


CBER Reference Immune Globulin,

Lot 176, for Measles and Polio antibody levels

Required anti-measles potency:  NLT 0.6 times the level of Lot 176 (42 IU/mL, recent data by a neutralization assay).

Required anti-polio potency: NLT 0.28 for Type 1, 0.25 for Type 2, or 0.20 for Type 3, times the level of Lot 176


Lot 176:  2 IU anti-HBs/mL 

              95 IU anti-HAV/mL

5, 6


CBER Reference Hepatitis B Immune Globulin, Lot 2


220 IU anti-HBs/mL


Albumin, PPF, IGIV, IGSC, and some specific IGIV products 

CBER Reference Prekallikrein Activator (PKA), Lot 3

100 IU PKA/mL

Required limit for PPF: NMT 35.7 IU/mL

Standard limit for IgG product:  NMT 35 IU/mL 3% IgG


Rho(D) IG, Rho(D) IGIV

Global potency standard for anti-D immune globulin: 

WHO 2nd IS (NIBSC 01/572)/Ph Eur 1st BRP/CBER Lot 4

285 IU anti-D /mL 


Standard dose for use in preventing HDN: NLT 1500 IU (300 mg)


7, 8

IGIV, IGSC,  specific IGIV products

WHO International Reference Reagents (IRRs) to standardize hemagglutination testing  for anti-D:

Positive IRR: NIBSC 02/228; CBER Lot 1A

Negative IRR: NIBSC 02/226, CBER Lot 1N-a 

Positive IRR:

0.0475 IU anti-D/mL 5% IGIV


Nominal titer of 8 by a direct hemagglutination method




Same as above

NIBSC/Ph Eur 1st BRP/CBER Reference reagents to set the maximum limit of anti-D in immune globulins: 


Positive RR: NIBSC 04/132; CBER Lot 1B


Negative RR:  NIBSC 04/140; CBER Lot 1N-b

Same titer as above except with more ampoules filled and shared among NIBSC, EDQM and CBER 


Maximum anti-D titer for 5% IgG :  NMT the level in Positive RR (by the direct hemagglutination method) 

10, 11


Abbreviations: IG, Immune Globulin (Human); IGIV, Immune Globulin Intravenous (Human); IGSC, Immune Globulin Subcutaneous (Human) ; HBIG, Hepatitis B Immune Globulin (Human); HBIGIV, Hepatitis B Immune Globulin Intravenous (Human); PPF, Plasma Protein Fraction (Human); BRP, Biological Reference Preparation; HDN, hemolytic disease of the newborn.


Safety Standards for In-Process Control:



Viral copies or IU per mL


Ref. #

CBER Parvovirus B19-positive plasma standard (genotype 1)


Use:  Validating B19 NAT methods used for testing plasma pools (minipools and manufacturing pools) as analytical procedures to ensure the quality of starting plasma pools for all plasma-derived products. 


106 genome equivalents (geq) or IU of B19 DNA per mL


CBER’s proposed maximum level of B19 DNA in the manufacturing pool: 

104 IU/mL




CBER  Hepatitis A Virus-positive human plasma standard


Use:  Validating HAV NAT methods used as described above.



1.2 x 104 copies/mL or 6000 IU/mL


No proposed limit of HAV RNA in the manufacturing pool yet.




Potency and Safety Standards under Development:


1.   In collaboration with Dr. Elaine Gray of NIBSC, the 4th International Standard for Factor IX Concentrate will be established.  


2.   In collaboration with Dr. Morag Ferguson of NIBSC, the 2nd International Standard for Anti-HBs Immunoglobulin/CBER Lot 3 will be established to replace both the 1st international reference preparation for anti-HBs Immunoglobulin (100 IU anti-HBs/mL when reconstituted) and CBER Reference Hepatitis B Immune Globulin, Lot 2.  Nabi has kindly provided a candidate 5% HBIG preparation.


3.   A new CBER Reference Immune Globulin, Lot 177, will be established to replace Lot 176.  Baxter has kindly provided a candidate 10% IGIV preparation. 


4.   In collaboration with Dr. Susan Thorpe of NIBSC and Dr. Marie-Emmanuelle Behr-Gross of EDQM, new international reference reagents will be established to standardize hemagglutination testing for anti-A and anti-B hemagglutinins and to define the maximum levels of both in immune globulin products.  Baxter has kindly provided a negative IGIV preparation derived from type AB plasma donations.


5.   In collaboration with Dr. Sally Baylis of NIBSC, a parvovirus B19 genotype panel containing all three B19 genotypes will be established.




1.   Report to Biological Standardization Steering Committee.  PA/PH/BIO (96) 18, R,   August 1996.

2.   Kirschbaum N, Wood L, Lachenbruch P, Weinstein M, Daas A, Rautman G, Spieser JM, Buchheit KH.  Pharmeuropa Bio 2002; 1: 31-60.

3.   Hubbard AR, Sands D, Chang AC, Mazurier C.  Thromb Haemost 2002; 88: 380-6.

4.   Whitton C, Sands D, Lee T, Chang A, Longstaff C.  Thromb Haemost 2005; 93: 261-6.

5.   21 CFR 640.104  Potency.

6.   Audet S, Virata-Theimer ML, Beeler J, Scott DE, Frazier DJ, Mikolajczyk MG, Eller N, Chen F, Yu MW.  J Infect Dis 2006; 194: 781-9.

7.   Thorpe SJ, Sands D, Fox B, Behr-Gross M-E, Schäffner G, Yu MW.  Vox Sang 2003; 85: 313-21.

8.   Thorpe SJ, Sands D, Fox B, Schäffner G, Yu MW, Behr-Gross M-E.  Pharmeuropa Bio 2003; 2: 9-26.

9.   Thorpe SJ, Fox B, Heath A, Dolman C, Virata ML, Yu MW, Thorpe R.  Vox Sang 2005; 88: 278-87.

10. Thorpe SJ, Fox B, Heath A, Behr-Gross M-E, Virata ML, Yu MW.  Biologicals 2006; 34:  209-12.

11. Thorpe SJ, Fox B, Heath A, Behr-Gross M-E, Virata ML, Yu MW.  Pharmeuropa Bio             2006; 1: 579-85.  

12. Saldanha J, Lelie N, Yu MW, Heath A, and the B19 Collaborative Study Group.  Vox Sang 2002; 82: 24-31.


13. Saldanha J, Heath A, Lelie N, Pisani G, Yu MY, and the Collaborative Study Group.   Vox Sang 2005; 89: 52-8.



C.  Joint FDA/WHO Minimum Potency Standards for Certain Blood Grouping

      Reagents, Sheryl Kochman, DBA, OBRR, FDA



Minimum Potency Standards for Blood Grouping Reagents


·         Blood Grouping Reagent Anti-A minimum potency standard – replaces FDA lot 6A1

·         Blood Grouping Reagent Anti-B minimum potency standard – replaces FDA lot 7A-11

·         Blood Grouping Reagent Anti-D minimum potency standard – replaces FDA lots 4a-1 and 92




1.         Thorpe SJ, Fox B, Heath AB, Scott M, de Haas M, Kochman S, Padilla A. 

            International standards for minimum potency of anti-A and anti-B blood grouping            reagents: evaluation of candidate preparations in an international collaborative           study.  Vox Sanguinis 2006; 91, 4: 336-344


2.          Thorpe, SJ.  Fox, B, Heath, AB, Scott M, de Haas, M, Kochman S, Padilla A.                   An International Standard for specifying the minimum potency of anti-D blood-       grouping reagents: evaluation of a candidate preparation in an international            collaborative study.  Vox Sanguinis 2006; 90, 2: 131-139.