Plasma fractionation

Human blood plasma contains over 700 different proteins (qv) (1). Some of these are used in the treatment of illness and injury and form a set of pharmaceutical products that have become essential to modem medicine (Table 1). Preparation of these products is commonly referred to as blood plasma fractionation, an activity often regarded as a branch of medical technology, but which is actually a process industry engaged in the manufacture of speciaUst biopharmaceutical products derived from a natural biological feedstock (see Pharmaceuticals). 

Product Clinical appHcation Molecularweight x 10 Normal plasma concentration, g/L 

Factor VIII Coagulation proteins hemophilia A treatment 300 3 X 10-  

Factor IX complex treatment of hemophilia B and other coagulation 57 5 X 10-  

History. Methods for the fractionation of plasma were developed as a contribution to the U.S. war effort in the 1940s (2). Following pubHcation of a seminal treatise on the physical chemistry of proteins (3), a research group was estabUshed which was subsequendy commissioned to develop a blood volume expander for the treatment of military casualties. Process methods were developed for the preparation of a stable, physiologically acceptable solution of alburnin [103218-45-7] the principal osmotic protein in blood. Eady preparations, derived from equine and bovine plasma, caused allergic reactions when tested in humans and were replaced by products obtained from human plasma (4). Process studies were stiU being carried out in the pilot-plant laboratory at Harvard in December 1941 when the small supply of experimental product was mshed to Hawaii to treat casualties at the U.S. naval base at Pead Harbor. On January 5, 1942 the decision was made to embark on large-scale manufacture at a number of U.S. pharmaceutical plants (4,5). 

---

A method for the fractionation of plasma, allowing albumin, y-globulin, and fibrinogen to become available for clinical use, was developed during World War II (see also Fractionation, blood-plasma fractionation). A stainless steel blood cell separation bowl, developed in the early 1950s, was the earhest blood cell separator. A disposable polycarbonate version of the separation device, now known as the Haemonetics Latham bowl for its inventor, was first used to collect platelets from a blood donor in 1971. Another cell separation rotor was developed to faciUtate white cell collections. This donut-shaped rotor has evolved to the advanced separation chamber of the COBE Spectra apheresis machine. 

Primary blood components iaclude plasma, red blood cells (erythrocytes), white blood cells (leukocytes), platelets (thrombocytes), and stem cells. Plasma consists of water dissolved proteias, ie, fibrinogen, albumins, and globulins coagulation factors and nutrients. The principal plasma-derived blood products are siagle-donor plasma (SDP), produced by sedimentation from whole blood donations fresh frozen plasma (FFP), collected both by apheresis and from whole blood collections cryoprecipitate, produced by cryoprecipitation of FFP albumin, collected through apheresis and coagulation factors, produced by fractionation from FFP and by apheresis (see Fractionation, blood-plasma fractionation). 

A. Boyum and co-workers, "Density Dependent Separation of White Blood Cells," inj. R. Harris, ed.. Blood Separation and Plasma Fractionation, Wiley-Liss, New York, 1991. 

Full details of this work were pubHshed (6) and the processes, or variants of them, were introduced in a number of other countries. In the United States, the pharmaceutical industry continued to provide manufacturing sites, treating plasma fractionation as a normal commercial activity. In many other countries processing was undertaken by the Red Cross or blood transfusion services that emerged following Wodd War II. In these organisations plasma fractionation was part of a larger operation to provide whole blood, blood components, and speciaUst medical services on a national basis. These different approaches resulted in the development of two distinct sectors in the plasma fractionation industry ie, a commercial or for-profit sector based on paid donors and a noncommercial or not-for-profit sector based on unpaid donors. 

In 1993 there were over 100 organisations undertaking plasma fractionation woddwide, having plant capacities ranging from 4 to 1800 m /yr. Virtually all of these plants use methods based on those originally devised. Table 2 Hsts the six commercial manufacturers in the United States and the largest plasma fractionators woddwide. 

Plasma fractionation is unusual in pharmaceutical manufacturing because it involves the processing of proteins and the preparation of multiple products from a single feedstock. A wide range of unit operations are utilized to accompHsh these tasks. They are Hsted in Table 3 some are common to a number of products and all must be closely integrated. The overall manufacturing operation can be represented as a set of individual product streams, each based on the processing of an intermediate product derived from a mainstream fractionation process (Fig. 1). 

Fig. 2. Typical plasma fractionation operation where Fr represents fraction. Based on processes at the Protein Fractionation Centre, Scottish National...

Inactivation and Removal of Viruses. In developing methods of plasma fractionation, the possibiHty of transmitting infection from human vimses present in the starting plasma pool has been recognized (4,5). Consequentiy, studies of product stabiHty encompass investigation of heat treatment of products in both solution (100) and dried (101) states to estabHsh vimcidal procedures that could be appHed to the final product. Salts of fatty acid anions, such as sodium caprylate [1984-06-17, and the acetyl derivative of the amino acid tryptophan, sodium acetyl-tryptophanate [87-32-17, are capable of stabilizing albumin solutions to 60°C for 10 hours (100) this procedure prevents the transmission of viral hepatitis (102,103). The degree of protein stabilization obtained (104) and the safety of the product in clinical practice have been confirmed (105,106). The procedure has also been shown to inactivate the human immunodeficiency vims (HIV) (107). 

Process Rationale. The products of plasma fractionation must be both safe and efftcaceous, having an active component, protein composition, formulation, stabiUty, and dose form appropriate to the intended clinical appHcation. Processing must address a number of specific issues for each product. Different manufacturers may choose a different set or combination of unit operations for this purpose. 

Estimates for a number of economic aspects of plasma fractionation can be made (200—206). The world capacity for plasma fractionation exceeded 20,000 t of plasma in 1990 and has increased by about 75% since 1980, with strong growth in the not-for-profit sector (Fig. 4). The quantity of plasma processed in 1993 was about 17,000 t/yr the commercial sector accounts for about 70% of this, with over 8000 t/yr in the form of source plasma from paid donors (Fig. 5). Plant capacities and throughput are usually quoted in terms of principal products, such as albumin and Factor VIII. These figures may not encompass manufacture of other products. 

Fig. 4. Capacity of plasma fractionation plants O, not-for-profit capacities , for-profit capacities in (—), the United States and (--------), Europe (—...

Worldwide Directory of Plasma Fractionators, Marketing Research Bureau, Laguna Beach, Calif., 1990. 

The free plasma fraction (fp) determines the relation between free, bound ( plasma binding PB%), and total concentration. 

The total clearance increases in proportion to the free plasma fraction while free plasma clearance remains constant. 

The apparent volume of distribution (Vd) slightly increases depending on plasma volume (Fp), tissue volume (Ft), and free tissue fraction (ft) whereas the half-life slightly decreases with significantly increasing free plasma fraction. 

---