1. Concurrent and Component Plasma: CBER Considerations on Standards and Labeling BPAC April 28, 2011 Mark Weinstein, Ph.D. Office of Blood Research and Review CBER, FDA

2. Issue FDA seeks to establish minimum standards for the collection, storage and freezing of Concurrent Plasma, a product intended for further manufacturing into injectable end products, and labeling that reflects these standards, in order to reduce variability of such plasma. Component Plasma, defined as outdated apheresis FFP as a sole collection, is known to meet the standards under consideration. 2

3. Issue- II For all conditions of plasma collection, rate of freezing, and storage temperature, manufacturers may request, under contract with blood collectors, conditions that are more stringent than the minimum FDA standards outlined here.

4. Topics CBER Considerations for the categories of Concurrent Plasma Rationale for Establishing Quality Standards Scientific evidence that supports categories under consideration Industry practices for the collection and use of recovered plasma Rationale for standards under consideration Labeling of Concurrent and Component Plasma

5. FDA Standards Source Plasma Recovered plasma Collection methodPlasmapheresis Whole blood or Whole blood or Apheresis Time from collection to freezing ImmediatelyUndefined Freezing conditions, temperature <-20  CUndefined Storage, expiration <-20  C, 10 yrUndefined Shipping temperature <-5  CUndefined Allowable deviation Can exceed -20  C for < 72 h total Never > -5  C, always frozenUndefined

6. Concurrent Plasma Categories Placement in <-20°C freezer LabileNon Labile Non Injectable 70 IU/100 ml* xxx <24 hr avg FVIII < 70 IU/100 ml* xx <72 hr xx >72 hr x 6 * process validation and possibly also Quality Control testing under GMP

7. RATIONALE FOR ESTABLISHING QUALITY STANDARDS 7

8. Elements of Plasma Quality Plasma quality is a relative term – Different for labile or non-labile products FVIII historically is a common indicator of plasma quality: – very labile – commercial interest in obtaining high-yield Plasma for injectable products should not contain substances that, if not removed or neutralized during the manufacturing process, could adversely effect the safety or efficacy of the final product. 8

9. Parameters Affecting Plasma Quality Whole Blood or Apheresis Plasma ACD, CPD Plasma Separation Method Surfaces Time Temperature Cell content Plasma unit Storage time, temperature FFP Cryoppt Cryosup Thaw Method Concurrent Plasma Rate of freezing

10. Plasma in contact with cellular components can undergo spontaneous proteolytic activity over time

11. Spontaneous proteolytic activity determined by S-2288 during platelet storage 22oC (Wallvik et al Transf Med 1992)

12. Coagulation factors, inhibitors, and other plasma components can change over time, becoming proteolyzed, aggregated, activated or inactivated in liquid plasma, leading to increased heterogeneity of constituents with poorly understood or controlled physical properties

13. Examples of Protein Activation or Inactivation Over Time Change in Factor XI and Protein S in thawed plasma (n = 18) prepared from FP24 over 5 day storage at 1-6 ◦ C. Day 0 = immediate post thaw 13 Data from Yazer et al, Transfusion, 2008

14. Percentage change in clotting factors in thawed plasma (n = 18) prepared from FP24 after 120 h storage at 1-6 ◦ C. Day 0 = immediate post thaw Data from Yazer et al, Transfusion, 2008 48:2525-2530

15. Activated factors could potentially become concentrated in certain plasma fractions leading to adverse events

16. The Cohn-Oncley Blood Plasma Fractionation Process

17. Plasma Types that Presently Can Be Distributed as Recovered Plasma 17

18. Plasma Definitions (CFR) Fresh Frozen Plasma [21 CFR 640.34 (b)] The plasma shall be separated from the red blood cells, and placed in a freezer within 8 hrs or within the timeframe specified in the directions for use for the blood collecting, processing, and storage system, and stored at <-18 o C Plasma, Cryoprecipitate Reduced: is prepared from FFP that is thawed and centrifuged, with the cryoprecipitate removed by centrifugation. 18

19. Plasma Definitions (CFR) cont. Platelet Rich Plasma 72 hr from time of collection of source blood, provided labeling recommends storage (20 – 24 o C or between 1 and 6 o C). 5 days if certain approved containers are used (20 to 24 o C) Liquid Plasma is separated no later than 5 days after the expiration date of the Whole Blood [expiration can be 21 or 35 days depending on anticoagulant]. Plasma may be stored at <-18 o C. Liquid plasma is stored at refrigerator temperature 1-6 o C. 19

20. Plasma Definitions ( AABB Circular) Fresh Frozen Plasma is prepared from whole blood or apheresis collection and frozen at -18°C or colder within the time frame as specified in the directions for use for the blood collection, processing, and storage system. Plasma frozen within 24 hours after phlebotomy: must be separated and placed at <-18 o C within 24 hours of whole blood collection. Thawed Plasma is derived from FFP and PF24 prepared in a way that ensures sterility (closed system), thawed at 30-37 o C, and maintained at 1-6 o C for 1-5 days. A product not in the CFR, not a licensed product. 20

21. Scientific Considerations for Standards for Concurrent Plasma 21

22. Considerations for establishing time from plasma collection to freezing, cell contact time, and FVIII yield for plasma used to produce labile versus non-labile products 22

23. Clotting Factors and Inhibitors, Markers of Activated Hemostasis and Proteolysis Plasma in freezer three hours after donation Clotting factors and inhibitors Apheresis plasma (n= 60) Whole blood plasma (n= 100) Factor VIII10084.5 Factor IX12096 Factor XI10787 Protein S8780 Coagulation activation and proteolysis markers F1+2 (nmol/L)0.330.42 PF426.5594 Neutrophil elastase (micrograms/L) 45161 Runkle et al, Transfusion 2005

24. Plasma Separated from Whole Blood and Placed in a Freezer within 24 Hours: Literature Review (references in issue summary) Time with cells (h) Time without cells temperatureFactor VIII (IU/100 ml) 81-6°C64 161-6 °C 822-24°C71 161-6°C 241-6°C76 ??1-6 °C72 18-201-6 °C55 16-2022 °C77 24 24 °C 75 24 20 °C 64 24

25. 25 Most studies suggest that holding Whole Blood for up to 24 hours from collection to freezing, under a variety of temperatures and cell contact time, will yield plasma containing an average Factor VIII level of >70 IU/100 ml. Other studies indicate that this is true of apheresis plasma as well.

26. Freezing Rate 26

27. Effect of Freezing of on FFP ( Akerblom, et al, 1992) Rapid freeze (-40 o C, <40 min): loss of about 8% FVIII from pre to post freeze; 9% increase in beta-TG (platelet lysis) but not statistically significant Slow freeze, (-20 o C): 20% loss FVIII; 8% loss FV; 14% increase in beta-TG Slow freeze, (-40 o C): 14% loss FVIII; freeze rate more important than final temperature No difference between fast and slow freeze in FVII, vWF, soluble fibrin, TAT complex, C 1 -esterase inhibitor

28. European Pharmacopoeia PurposeLabile proteins for fractionation Non labile protein for fractionation Collection method Plasmapheresis or Plasma from whole blood Plasmapheresis Plasma from whole blood Time from collection to freezing <24 hrs <72 hrs Freezing conditions, temperature Frozen to a core temp of <-25  C in <12 hr Chamber at <-20  C FVIII Average FVIII >70 IU/100 ml in QC testing under GMP Storage <-20  C Shipping temperature <-20  C Allowable deviation Exceeds -20  C not more than 72 h total One time > -15  C, Never > -5  C

29. Summary data on FVIII activity recovery at different plasma freezing conditions (Bravo, 2006) Chamber at -30°C-25°C-20°C FVIII:C recovery Mean + SD (%) 8585, 8475 FVIII recoveries in the cryoprecipitate at different plasma freezing conditions Temperatures -30°C-25°C-20°C FVIII in cryoprecipitate (IU FVIII/l plasma) 464447476

30. Rapid freezing increases FVIII yield in thawed plasma compared to slower freezing but may not increase FVIII yield in cryoprecipitate or the final yield of other plasma proteins made under various manufacturing conditions

31. FDA is not considering specifying the rate of freezing in establishing minimum quality standards for Concurrent Plasma.

32. Storage Freezing Temperature and Time of Storage 32

33. Data to Show No Change in FVIII and Other Proteins in FFP Stored at –20 o C vs – 40 o C for 36 Months Kotitschke, 2000

34. INDUSTRY PRACTICE 34

35. Industry Practice 2004 <8 hr< 24 hr<72 hr<120 hr Grifols IGIV, Albumin CSLB FVIII, vWF, Albumin, IGIV Albumin, IGIV Octapharma SD PlasmaFVIII, Albumin, IGIV Albumin, IGIV Bayer (now Talecris) Recovered plasma used only to make Fraction IV-1 paste for Alpha1 PI; time to placement in freezer not given ARC Plasma placed in freezer within 8 and 24 hr, but products made from 8 and 24 hr plasma were not identified 35

36. Standards Under Consideration for Concurrent Plasma To Be Used for Injectable Products 36

37. Concurrent Plasma Category Examples of injectable product Time from donation to placement in freezer Freeze Store and Ship Labile and Non-labile products FVIII, vWF, IGIV, Albumin, (Average FVIII >70 IU/100 ml, by process validation and possibly QC testing under GMP *) <24 hr Freezer at < -20◦C < -20 ◦C Non-labile products only IGIV, Albumin<72 hr Freezer at < -20 ◦C < -20 ◦C 37 * Possible exception for FFP

38. Labile products made only from plasma held for < 24 hours, at specified temperatures, from collection to placement in freezer. Rationale: Reflects available data and is in accordance with current US industry practice. Non-labile products made from plasma held for < 72 hours, at specified temperatures, from collection to placement in freezer. Rationale: Addresses concern about increase in protein heterogeneity over time. Within current practice of most sponsors.

39. Plasma that has been thawed and refrozen would not be used for injectable products. Rationale: There is a concern about the quality of plasma thawed and refrozen multiple times based on potential for changes in some proteins. Source Plasma has similar limitations. Replace <-20°C for <-18°C for all conditions of freezing, shipping and storage Rationale: The temperature limit of <-20°C for all conditions is the current industry practice and is supported by data (Kotitschke).

40. Allowable shipping and storage temperature deviation can exceed - 20°C for < 72 hours total, never greater than -5°C (never refrozen). Rationale: The current FDA standards (CFR) for Source Plasma are known to assure protein stability. Storage up to 3 years from collection. Rationale: Data (Kotitschke) support the stability of plasma held at <-20°C for this time period.

41. Average FVIII level > 70 IU/100 ml for plasma used to make labile products as process validation and possibly also Quality Control testing under GMP. Rationale: This GMP quality control requirement is the same as that in the European Pharmacopoeia. The FVIII activity of >70 IU/100 ml for whole blood and apheresis plasma is an achievable benchmark, a desirable level of activity for manufacturing purposes, and is one marker of plasma molecular integrity.

42. European Pharmacopeia Guidelines for the Preparation of Plasma for Manufacturing “It is not intended that the determination of … factor VIII [is] to be carried out on each unit of plasma. [It is] rather given as [a guideline] for good manufacturing practice, the test for factor VIII being relevant for plasma intended for use in the preparation of concentrates of labile proteins. “The aim of good manufacturing practice is to conserve labile proteins as much as possible.”

43. European Pharmacopeia Guidelines for the Preparation of Plasma for Manufacturing Factor VIII. “Carry out the test using a pool of not fewer than 10 units. Thaw the samples to be examined, if necessary, at 37°C. Carry out the assay of factor VIII using a reference plasma calibrated against the international standard for human coagulation factor VIII in plasma. The activity is not less than 0.7 IU /ml.”

44. Considerations for Labeling of Concurrent and Component Plasma as Related to Process Control 44

45. Labeling for Concurrent Plasma Injectables For labile products: – “Caution: For Manufacturing Use Only. Placed in freezer < 24 hours. May be used for labile or non-labile injectable products." – (Note: Process control would assure an average FVIII >70 IU/100 ml) For non-labile products: – “Caution: For Manufacturing Use Only. Placed in freezer < 72 hours. May be used only for non-labile injectable products.“* *Same label for plasma: freezer <24 hr but FVIII <70 IU/100 ml

46. Labeling for Non-Injectable Use "Caution: For Use in Manufacturing Noninjectable Products Only.“ Frozen at >72 hr

47. Labeling for Component Plasma Injectables For labile products: – “Caution: For Manufacturing Use Only. Placed in freezer < 8 hours. May be used for labile or non- labile injectable products.“ Other labeling for apheresis products may be possible in the future.

48. Summary - I Currently the storage/freezing conditions for recovered plasma vary widely and are specified in short supply agreements between collectors and fractionators or manufacturers of IVDs. Variations in freezing, thawing, time in contact with cells and time in the liquid state affect the preservation, activation and inactivation of known and unknown plasma proteins, which can potentially affect the consistency and safety of finished products The manufacturing conditions specified for Concurrent Plasma will help to assure the manufacture of safe and effective plasma products. Component Plasma, i.e. apheresis FFP, already meets these standards. These candidate specifications are generally consistent with current manufacturing practices.

49. Summary – II Standardized categories for labeling of Concurrent Plasma and Component Plasma for key conditions of storage and freezing, will inform purchasers in the US and abroad about the salient characteristics of each product and establish minimum standards of processing and handling Standards for Concurrent Plasma will enable FDA to provide regulatory oversight that will increase consistency and limit the extent to which plasma with unknown or inferior properties could be used for manufacturing. Other categories may be established based on further evidence showing that safe and effective products can be produced from plasma manufactured under specified conditions.

50. Question 1. Please comment on FDA’s current considerations on standards for Concurrent Plasma – Labile products made only from plasma held at < 24 hours from collection to placement in freezer. – Average FVIII level > 70 IU/100 ml for plasma used to make labile products as Quality Control testing under GMP where appropriate (process validation). – Non-labile products made from plasma held at < 72 hours from collection to placement in freezer.

51. Question 1. (cont.) Please comment on FDA’s current considerations on standards for Concurrent Plasma – Replacing <-20°C for <-18°C for all conditions of freezing, shipping and storage – Allowable shipping and storage temperature deviation can exceed - 20°C < 72 hours total, never greater than -5°C (never refrozen). – Storage up to 3 years. – Plasma that has been thawed and refrozen would not be used for injectable products (with the possible exception of cryo-poor plasma).

52. Q2 A. Does the Committee agree with FDA’s current considerations on labeling categories for Concurrent Plasma? * process validation and possibly also Quality Control testing under GMP Placement in <-20°C freezer LabileNon Labile Non Injectable 70 IU/100 ml* xxx <24 hr avg FVIII < 70 IU/100 ml* xx <72 hr xx >72 hr x

53. * FVIII process validation and possibly also Quality Control testing under GMP Q2 B. Does the Committee agree with FDA’s current considerations on labeling categories for Component Plasma? Placement in <-20°C freezer LabileNon Labile Non Injectable <8 hr xxx Future products?*