IgG, IgA and IgM antibody preparations, method of making and method of use in treatment
An antibody preparation suitable for intravenous administration in humans includes IgG, IgA and at least 5% IgM antibodies by weight of the total amount of antibodies. The preparation is prepared from human plasma, has specific complement activating activity, and, in an in vitro assay with human serum suitable to determine the ability of the antibody preparation to activate complement unspecifically, the antibody preparation generates substantially no C5a and/or substantially no C3a. The antibody preparation can have medical uses.
1. An antibody preparation suitable for intravenous administration in humans comprising immunoglobulins IgG, IgA and IgM, wherein at least 15% by weight of the total immunoglobulins are IgM, at least 5% by weight of the total immunoglobulins are IgA and at least 40% by weight of the total immunoglobulins are IgG, wherein the antibody preparation is virus safe with respect to enveloped and non-enveloped virus, has an anticomplementary activity of ≤1 CH 50/mg protein, is stable in liquid form for at least 2 years when stored at 2 to 8° C., wherein the antibody preparation comprises a protein concentration of 40 g/L to 80 g/L and an immunoglobulin content of greater than 95% of the total protein content.
2. The antibody preparation of claim 1 having a proteolytic activity of less than 8 U/L.
3. The antibody preparation of claim 1 , wherein the antibody preparation is prepared from human plasma.
4. The antibody preparation of claim 1 wherein the immunoglobulins have not been modified by contacting with β-propiolactone.
5. The antibody preparation of claim 1 comprising less than 2% aggregates of 1200 kDa or above of the total immunoglobulin content as determined by high performance size exclusion chromatography.
6. The antibody preparation of claim 5 comprising less than 1.5% aggregates of 1200 kDa or above.
7. The antibody preparation of claim 1 wherein the anticomplementary activity is less than 0.75 CH50/mg protein.
8. The antibody preparation of claim 1 , wherein the antibody preparation is prepared in the absence of a step of heat treatment at a temperature 40° C. or above for more than 10 minutes.
9. The antibody preparation of claim 1 comprising a buffer comprising glycine and a pH in the range of pH 4 to 5.5.
10. The antibody preparation of claim 1 , wherein the antibody preparation is prepared from human plasma by a process comprising the steps of:
(a) preparing from the human plasma a plasma fraction as a solution containing immunoglobulins;
(b) mixing a C7 to C9 carboxylic acid with the solution and treating the mixed solution with a vibrating agitator to precipitate contaminating proteins;
(c) separating the precipitated proteins from the solution to yield an IgM-containing immunoglobulin composition;
(d) incubating the IgM-containing immunoglobulin composition at between pH3.5 and pH 4.5 to form an incubated solution;
(e) irradiating the incubated solution with UVC to form a UVC-irradiated solution; and
(f) filtering the UVC-irradiated solution under sterile conditions to form the antibody preparation suitable for intravenous administration in humans.
11. The antibody preparation of claim 10 wherein the carboxylic acid is octanoic acid.
12. The antibody preparation of claim 10 wherein the process further comprises subjecting the incubated solution obtained from step (d) to nanofiltration prior to said irradiating step (e).
13. The antibody preparation of claim 1 wherein at least 90% of the antibodies in the preparation are biologically active.
14. The antibody preparation of claim 1 wherein in an in vitro Rubella antigen-based assay suitable to determine Fc function, the activity of the Fc part of the antibodies of the antibody preparation is the same as that of a biological reference preparation ±10%.
15. A method of producing an antibody preparation according to claim 1 from human plasma comprising the steps of:
(a) preparing from the human plasma a plasma fraction as a solution containing immunoglobulins;
(b) mixing a C7 to C9 carboxylic acid with the solution and treating the mixed solution with a vibrating agitator to precipitate contaminating proteins;
(c) separating the precipitated proteins from the solution to yield an IgM-containing immunoglobulin composition;
(d) incubating the IgM-containing immunoglobulin composition at between pH3.5 and pH 4.5 to form an incubated solution;
(e) irradiating the incubated solution with UVC to form a UVC-irradiated solution; and
(f) filtering the UVC-irradiated solution under sterile conditions to form the antibody preparation suitable for intravenous administration in humans.
16. The method of claim 15 wherein the C7 to C9 carboxylic acid is octanoic acid.
17. A method of treating a patient suffering from a bacterial infection or an immunological disorder comprising administering the antibody preparation of claim 1 .