JPS63194667A - Plasma treatment method and product thereof - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides a method for processing human plasma prior to administration to a patient;
More specifically, it is a method of removing blood type antibodies to avoid incompatibility between blood donors and recipients of different blood types.

Blood is a heterogeneous system in which various blood cells are suspended in an aqueous medium-pressure colloid suspension (plasma) consisting of proteins, riboproteins, and lipids, and organic metabolites are dissolved (-).Blood cells include red blood cells, white blood cells, and platelets. Plasma is the solution that remains after blood cells are removed from the blood. Plasma contains various proteins, which are produced by white blood cells as a result of the immune response to the presence of antigens in the blood. It contains secreted antibodies, proteins involved in other essential physiological processes, and coagulation factors necessary for hemostasis.

The major blood types of humans can be determined by the ABH antigen present on the surface of red blood cells. These antigens are oligosaccharides that bind to glycolipids or glycoproteins in the plasma membrane of red blood cells. There are three types of antigen structures: A'', ``B'', and non-A
``Non-``B'' (i.e. ``H'' was previously “O”)
It is called. A antigen, B antigen.

Alternatively, both A and B antigens may be present on the surface of each individual's red blood cells. Each individual's blood will contain natural antibodies (primarily IgM) against ABH antigens that are not present on their own red blood cells. The conditions are summarized in Table 1 below.

Table 1 Blood type Antigen on red blood cells Antibodies in blood AA Animal substance Anti-B BB substance Anti-A AB Both A and B substances Neither anti-A nor anti-B H
It is O non-A non-B substance anti-A and anti-B (H substance)
People who have sustained a trauma that results in bilateral blood loss often require transfusions of blood or plasma to compensate for the blood loss. Plasma is often used in place of whole blood because it is easier to store.

When transfusing plasma, an incompatibility between donor and recipient occurs whenever the donor's blood (and therefore the plasma) contains antibodies against the ABH antigen (if present) present on the recipient's red blood cells. occurs.

For example, plasma from a donor with type A blood cannot be transfused to a recipient with type B blood. This is because the donor's plasma contains anti-B antibodies that are thought to react with the recipient's blood. Therefore, plasma from a donor with type H blood (
(including both anti-A and anti-B antibodies) are incompatible with recipients with blood types other than H. Plasma from a donor with reverse KAB blood (contains neither anti-A nor anti-B antibodies)
is compatible with any recipient regardless of blood type.

Because of the possibility of incompatibility, traditionally each type of plasma had to be carefully separated and stored separately to prevent the plasma from being transfused to an incompatible recipient. It would be desirable to provide a process to remove antibodies to incompatible antigens from plasma, in order to provide a 'universal' plasma that can be administered to any recipient regardless of blood type. However, such a process The method must be extremely specific to ensure that essential blood proteins (particularly clotting factors) are not removed or reduced to inactive proportions.Currently, there are at least 15 known clotting factors. Interference with the activity of any of these factors in blood plasma will reduce the recipient's blood's ability to clot. This is a serious problem since it is necessary that the coagulation function of the blood is not impaired.

It is known to employ immunoadsorption to specifically remove anti-A and anti-B antibodies from the blood of a loved one who is about to receive a bone marrow transplant.

Benzinger et al. (1981), “An immunoadsorption method for removing type A and type B antibodies”, N. Engl., J. Med.

304:160-162; Benzinger (1981)
, Artificial Organs 5:254-258
; Benzinger et al. (1981), ) Lance Fusion 21:335-342; and Benzinger et al.
82), Progress in Clinical and Biological Research 88:295-300.

The present invention processes human plasma obtained from any blood donor and
Plasma containers that are compatible with any recipient, regardless of the recipient's blood type, provide a method for making blood products. The method consists of passing plasma from a donor through an immunoadsorption zone capable of binding one or more of the above-mentioned antibodies to remove them.

This immunoadsorption zone contains receptors specific for both anti-A and anti-B antibodies, generally anti-(anti-A) and anti-
The (anti-B) antibody or the A and B antigens themselves are immobilized therein and contained in the t-state.

This method requires that the ViABH antibody be substantially removed from the complete preparation at a dilution of 1:2, preferably at zero dilution, at least to levels detectable by the standard antiglobulin (Coombs) test.

Unexpectedly, it has been found that this treatment does not substantially reduce the concentration of other blood proteins (particularly clotting factors) necessary for the biological function of the plasma in the recipient.

The present invention provides a source of human plasma derived from a donor of any blood type that can be transfused to a recipient regardless of blood type.

This method can be performed in hospitals or blood banks;
The need to separate plasma by blood type is avoided in order to preserve this dense plasma from being administered to recipients with incompatible blood types. This method can also be used in emergency situations, such as when there is a limited number of blood donors available and when plasma cannot be used for a particular individual in need of a blood transfusion without a way to dispose of this food supply.

The method of the present invention uses an immunoadsorption zone in which at least anti-A antibody or anti-B antibody-specific receptors, particularly receptors for both of these antibodies, are immobilized. By passing human plasma through this immunoadsorption zone, antibodies associated with blood group incompatibility are virtually completely removed, and the plasma can be administered to any recipient regardless of the recipient's blood type. can.

The synthetic receptor is a natural or synthetic compound that can specifically bind anti-A and anti-B antibodies and form an immunoadsorption zone as a permeable medium with fixed internal pressure. Particularly useful are antibodies formed against natural type A and BW blood cell surface antigens, and both anti-A and anti-B antibodies, i.e., anti-(anti-A) and anti-(anti-B).
It is an antibody. These antibodies can be obtained from various mammals using conventional methods.

For example, it can be obtained by hyperimmunizing rabbits, sheep, macus, or goats, and then using the resulting antiserum. Alternatively, monoclonal antibodies can be found in Köhler and Milstein (197, 15) Nature 356
:495-497, and can be produced by the ratio method. Techniques for producing antibodies of this type are well known in the art and do not form part of the present invention.

Preferred receptors for the present invention are those described in Chem, Soc, Rev.
, ), pp. 423-452, human blood group A and B antigens are artificially synthesized by the method described.

2. Solid phase immunoadsorbent The immunoadsorption zone of the present invention consists of a receptor immobilized on a solid phase immunoadsorbent. Solid phase materials of this type can be selected from a wide variety of materials, typically silicates or water-insoluble polymers used to pack separation columns, or water-permeable membranes.

Generally, the immobile solid phase of the present invention must be capable of covalently or non-covalently binding specific receptors for anti-A and anti-B antibodies, and must be biologically compatible with human blood. Must be.

Suitable materials include silica gel, glass beads, cross-linked dextran, polyacrylamide, and other biologically inert materials that can be derivatized (generally amino ratio) to a fixed ratio of receptors of interest. .

3. Immobilization of the receptor in the immunoadsorption zone 1 and the receptor can be immobilized in the immunoadsorption zone by various general methods, depending in particular on the nature of the binding material. Generally, one or more functional groups on an immunoadsorbent must be modified to enable binding to a receptor. The covalent attachment of proteins to immunoadsorbents (particularly antibodies) is taught in US Pat. Nos. 3,555,145 and 4,108,974.

For example, an artificial antigen (V mini-1 supra) contains a cricoside union to 8-methoxycarbonyloctyl alcohol, the former acting as a tight bridging arm for attachment to a solid support.

This antigen is reacted with an acyl hydrazide to form an acyl azide, which is then aminated to form a solid support, and
is reacted with a suitable amine on the solid material.

B. Plasma Processing 1. Separation of Plasma from Whole Blood Plasma is obtained by separating red blood cells, white blood cells and platelets from whole blood in a well-known manner. Once the plasma is obtained, it may be processed directly or it may be stored and processed at a later time.

2. Introduction of plasma into the immunoadsorption zone Before introducing plasma into the immunoadsorption zone, the immunoadsorption column or membrane is preferably treated to prevent non-specific protein adsorption. This is done by washing the immunoadsorption zone with a colloidal xylene solution (Collodion) or with a saline solution containing 1% human serum albumin, and incubating the zone with the washing solution preferably overnight at 4°C. Can be done. The column is then washed with saline before processing the plasma.

Final concentration of anti-A and anti-B antibodies at maximum dilution of 1:2
The plasma is passed through the immunoadsorption zone at a specific velocity chosen such that it is undetectable by a standard antiglobulin (Kumuse) test, preferably at zero dilution. The standard antiglobulin (Kumuse) test is described by Morrison (1972), “
Blood Transfusion in Clinical Medicine” (Blackwell Scientific Publications, Oxford)
It is described on pages 420-428.

After this treatment, the plasma column can be stored or the plasma fraction can be used in a conventional manner.

In the case of blood products, particularly anti-hemophilic factors, active]hyprothrombin complexes, etc., it is often desirable to further process the product to prevent degradation, such as by lyophilization.

The following examples are presented for illustrative purposes only and are not intended to be limiting.

EXAMPLES MATERIALS AND METHODS Manufactured as described by Lemieux (supra) K. Carbohydrate-based antigens (B-trisaccharides) were prepared on crystalline silica (Synsolve A) and non-Hysilica (Chem Biomed Inc., Edmonton, Canada) according to the manufacturer's instructions. K-conjugated vomit. 1 g of silica containing 10 micromoles of bound antigen was placed in a cartridge suitable for conducting plasma.

Plasma was obtained from 10 normal, healthy blood donors who had not taken any medication for at least 14 days. Plasma (1014) from each donor was incubated for 5 minutes and applied to a fresh cartridge, and post-process samples were collected for analysis.

Results Both processed and unprocessed plasma from each donor was tested for anti-B
Test and ratio for antibody titer. The results are shown in Table 1. Antibody removal is very effective, with a titer of 1:2 after treatment.
Atsushi below.

Table 1 1 1:64 1:2 2 1:32 0 3 1:32 0 4 1:16 0 5 1:16 CJ 6 G16 0 7 1:16 1:2 8 1:8 0 9 1:8 0 10 1:80 Coagulation parameters were also measured as shown in Table 2. The t values shown are the average for all 10 donors. Prothrombin time is the time required to convert prothrombin to thrombin.

No significant deterioration was observed in this parameter. PTT is partial thromboplastin time. A slight increase in this parameter was observed in 11*, which is considered to be due to partial adsorption of 2 or more coagulation factors9, but this increase is not clinically significant. Similarly, reductions in factor Ⅰ (anti-hemophilic factor), factor V (proactilein) and fibrinogen may also be due to non-specific adsorption, but are well within the limits of useful plasma.

Table 2 PTT (seconds) 2Z1±2.0 40±7 Factor V (%) 95± 3 82± 4 Factor ■
125±5 75±10 Table 6 shows the concentration of immunoglobulin and albumin to globulin ratio in plasma before and after treatment. Since anti-(ABH antigen) antibodies are primarily IgM, a decrease in IgM levels (and to a lesser extent IgG levels) can be expected. A decrease in immunoglobulin may also be associated with an increase in the albumin/globulin ratio.

Table 3 Parameter Before treatment After treatment IgG (m, y/dA') 800 ± 75
801 ± 19I gA (m, g/dl) 12
2± 12 158± 14I gM (mg/dj?
) 205±8 94± 11 Albumin/Globulin 1.46±, 08 1.75±, 1
Table 2 shows the results of quantitative electrophoretic measurements of five specific proteins and total protein in plasma both before and after treatment. Average values for all 10 donors are shown. Since this treatment aims to remove immunoglobulin, as expected, significant changes were observed only in the reduction of immunoglobulin.

Table 4 Protein Before treatment After treatment Total protein (E/di) 6.9 ± 1.3
6.6 ± 0.2 albumin (fi/dl)
4.01 ± 0.31 4.11 ± 0
．． 29 Table 5 shows the known activity ratio factor (adenosine diphosphate (
Figure 3 shows the effect of both pre- and post-treatment plasma on platelet aggregation and serotonin release in the presence of ADP), epinephrine, T50max, and collagen).

The effect of this treatment on these essential clotting functions is not significant.

Table 5 Blood/j-J & agglutination (%) Before treatment After treatment 5.ADP
85 ± 585 ± 5 1:100 epinephrine 92 ± 2 95 ±
6T50 max 81 ± 481 ± 3 Collagen 78 ± 4 81 ± 2 Serotoni/Hota (%) 5mmADP 18 ±
2 19 ± 21:100 epinephrine
25 ± 3 24 ± 2T50 max
19 ± 2 21 ± 3 Collagen 42 ± 4 44 ± 3 Therefore,
A method of rigorous plasma processing is provided that substantially completely removes anti-(ADH antigen) antibodies from plasma prior to transfusion. After this treatment, plasma can be administered to recipients regardless of their blood type. Unexpectedly, plasma subjected to this treatment retains virtually all of the other blood proteins measured from a clinically relevant point of view, which makes the plasma sufficiently bioactive to be used in the recipient's life. Although the present invention has been described in some detail by way of explanation and examples in order to provide a clear understanding of the invention, it is clear that certain changes and modifications may be made within the scope of the claims. Probably.

(5 other people)

Claims (9)

[Claims] (1) A method of treating human plasma from a donor before transfusion to a recipient to prevent ABH blood group incompatibility regardless of the blood type of the recipient, the method essentially comprising anti-A antibodies. The method uses an immunoadsorption zone containing immobilized receptors consisting of receptors specific for both anti-A and anti-B antibodies, and the method can detect virtually all anti-A and anti-B antibodies. is removed from the plasma, while other blood proteins are not removed. (2) A method of processing human blood to produce plasma suitable for transfusion to a recipient of any ABH blood type, the method essentially producing both anti-A and anti-B antibodies. The method uses an immunoadsorption zone containing a receptor comprising a receptor capable of binding, and the method separates substantially all blood cells and platelets from blood into plasma; substantially all anti-A antibodies and Anti-B antibodies are removed,
The plasma is passed through an immunoadsorption zone while other blood proteins are not removed; and the plasma is collected to obtain a source of plasma suitable for transfusion into a recipient, regardless of the recipient's blood type. The way it is done. (3) A method of transfusing plasma to a recipient regardless of ABH blood type incompatibility, the method comprising a receptor consisting essentially of a receptor capable of binding both anti-A and anti-B antibodies. The method uses an immunoadsorption zone to remove substantially all anti-A and anti-B antibodies from plasma without reducing the concentration of blood factors below the level required for clotting in the transfusion recipient. a method comprising: passing plasma through an immunoadsorbent zone under conditions in which the plasma is absorbed; and transfusing the plasma into a patient. (4) Claims 1, 2 or 2, wherein both anti-A and anti-B antibodies are removed at a 1:2 dilution to concentrations undetectable by the standard antiglobulin (Coombs) test. The method described in Section 3. (5) The method according to claim 1, 2 or 3, wherein the receptors of the immunoadsorption zone contain A antigen and B antigen. (6) The receptors in the immunoadsorption zone are anti-(anti-A) and anti-
The method according to claim 1, 2 or 3, comprising an (anti-B) antibody. (7) The method of claim 1, 2 or 3, wherein the immunoadsorption zone consists of a receptor immobilized on an insoluble biocompatible solid support. (8) The method according to claim 1, 2 or 3, wherein the immunoadsorption zone consists of a receptor immobilized in a permeable membrane. (9) Claims 1 and 2, wherein the blood factor is selected from the group consisting of factor VIII, factor V, fibrinogen, albumin, α-1-globulin, α-2-globulin, and β-globulin. The method described in Section 3 or Section 3.