Drug – bio-affecting and body treating compositions – Immunoglobulin – antiserum – antibody – or antibody fragment,... – Reduced antigenicity – reduced ability to bind complement – or...
Patent
1999-04-07
2000-10-24
Saunders, David
Drug, bio-affecting and body treating compositions
Immunoglobulin, antiserum, antibody, or antibody fragment,...
Reduced antigenicity, reduced ability to bind complement, or...
5303905, 5303871, 435264, C12N 950, A61K 39395, C07K 1600
Patent
active
06136312&
DESCRIPTION:
BRIEF SUMMARY
This application claims priority to PCT/CH97/00388, filed Oct. 14, 1997.
This invention relates to a method of producing an immunoglobulin solution suitable for intravenous application. Used as the starting material is a protein fraction obtained from human or animal blood which contains the immunoglobulins in concentrated form.
As is well known, immunoglobulins play an important role in the immune system of man and mammal in fighting off infections. The immunoglobulins are divided up into different classes (e.g. IgG, IgA, IgM, IgD and IgE) with differing biochemical and physiological properties. Until 1980 only IgG was isolated and used as an IV-well-tolerated product for prophylaxis and therapy. In EP-A-0 013 901, EP-A-0 413 187 and EP-A-0 352 500 IgM preparations are described, which have been made intravenously well tolerated mainly through treatment with .beta.-propiolactone. EP-A-0 413 188 describes a method in which the IV-tolerance is achieved through anion exchange chromatography with selective elution of the IV-tolerant fraction.
Object of the present invention is the production of a highly purified, IgM concentrate suitable for intravenous administration for therapy and prophylaxis. The product should have a low anticomplementary activity (ACA), and demonstrate a low blood pressure drop in a rat model, but the IgM molecules should not be chemically modified, however. This object was obtained surprisingly through treatment of a IgM-containing immunoglobulin solution with a protease.
The subject matter of the invention is therefore the method defined in claim 1.
The protease treatment is preferably an incubation at raised temperature in the presence of pepsin, papain, plasmin or thermolysine. The proteases can also be chemically modified, immobilized on a substrate and/or produced through genetic engineering. The preparation according to the inventive method can be transferred into an IV-administrable solution. Such a solution displays a reduction of the ACA, of the blood pressure drop in a rat model and of the C1q binding activity.
Suitable as starting materials for the method according to the present invention are immunoglobulin-containing solutions, such as, e.g. plasma, precipitate A or B from Kistler-Nitschmann-fractionation; Cohn fraction I/II/III; II/III; III; or other IgM-containing plasma fractions from human or animal plasma. For example, an immunoglobulin-containing fraction, such as precipitate B according to Kistler-Nitschmann, can be dissolved in a buffer, most of the impurities being removed through a precipitation with 0.5 to 5% octanoic acid at pH 4 to 6, preferably pH 5. Afterwards the solution is incubated at low ionic strength for 1 to 48 hours, preferably 9 hours, at a temperature of 20 to 50.degree. C., preferably 37.degree. C. with addition of at least 50 U/g of pepsin, preferably 600 U/G.
For further purification, the solution can be subjected to an adsorption, for example with a gel containing a DEAE-group in batch or column method. If the IgM concentration in the end product is supposed to be increased further, the IgM solution is put on an ion exchanger (e.g. TMAE-Fraktogel.RTM.). Through a selective elution, e.g. by means of a salt gradient or pH gradient, the IgM fraction can be isolated. Through ultrafiltration and diafiltration, for example a gel filtration, the solution can be concentrated and the electrolyte content can be adjusted to a final, intravenously well tolerated formulation. The protein concentration can amount to 1 to 20%, preferably 3 to 6%. The product can contain in addition proteins, preferably albumin, as well as sugar, preferably glucose or sucrose, or amino acids.
To assess the intravenous compatibility of immunoglobulin preparations, the anticomplementary activity (ACA) is usually used. To determine the ACA, a defined quantity of the product to be tested is incubated with a defined quantity of guinea pig complement and the remaining quantity of complement titrated. The ACA is indicated as consumption of CH50 per g of immunoglobulin. The indicated
REFERENCES:
patent: 4075193 (1978-02-01), Campbell et al.
patent: 5075425 (1991-12-01), Kotitschke et al.
Schiff P. et al Australian Paediatric Journal, 4:121-126 1968.
Anonymous, Pierce Catalog, pp. T67-Y70, Pierce Chemical Company, USA 1994.
P. Schiff et al, Australian Pediatric Journal, vol. 4, "The preparation, testing, and properties . . .", pp. 121-126, 1968.
J. T. Sgouris, VOX Sanguinis, vol. 13, No. 1, The preparation of plasmin treated immune . . . , pp. 71-84, Jul. 1, 1967.
M. M. Mayer, Experimental Immunochemistry, 2.sup.nd Edition, "Complement and Complement Fixation", pp. 133-240, 1961.
P. Spath et al, Scand. J. Immunol., vol. 18, "An Extended C1q-Binding Assay Using Lactoperoxidase-. . . ", pp. 319-328, 1983.
W. k. Bleeker et al, VOX Sanguinis, vol. 52, "An Animal Model for the Detection of Hypotensive . . . ", pp. 281-290, 1987.
DeCloux Amy
Rotkreuzstifung Zentrallaboratorium Blutspendedienst SRK
Saunders David
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