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Preparation method and application of immunoglobulin antibody with high sialic acid content

An immunoglobulin and sialic acid technology, applied in the direction of antibodies, anti-inflammatory agents, bone diseases, etc., can solve problems such as side effects, and achieve the effect of reducing side effects

Inactive Publication Date: 2012-02-01
XIAMEN UNIV
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Problems solved by technology

And it is precisely that the intravenous immunoglobulin G containing less than 5% of sialic acid is an effective ingredient for treating anti-inflammation (Yoshikatsu Kaneko, FalkNimmerjahn, Jeffrey V.Ravetch, Anti-Inflammatory Activity of Immunoglobulin G Resulting from Fc Sialylation, Science 313 , 670(2006)), large doses of intravenous immunoglobulin G can induce severe side effects in humans

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  • Preparation method and application of immunoglobulin antibody with high sialic acid content

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[0014] The following embodiments will further illustrate the present invention in conjunction with the accompanying drawings.

[0015] In 100ml of buffer solution (100mmol phosphoric acid solution, pH 7.5, containing 2mmol divalent manganese ions and 5mmol divalent magnesium ions), add intravenous immunoglobulin G (0.5g), uridine diphosphate-glucose (0.2g), Cytidine-5'-triphosphate (0.2g), sialic acid (0.1g), uridine diphosphate-galactose-4-epimerase enzyme (20 units activity), β1,4-galactosyltransferase ((20 units of activity), cytidine-5'-phosphate-sialic acid synthase (20 units of activity) and α2,6 sialyltransferase (20 units of activity). The reaction solution was stirred slowly at 4°C for 12h. The reaction solution Remove all glycosyltransferases through the His-Tag protein purification column. Then the reaction solution is dialyzed to remove unreacted small molecules such as uridine diphosphate-glucose, buffer solution and metal ions to obtain intravenous immunoglobulin...

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Abstract

The preparation method and application of an immunoglobulin antibody with high sialic acid content relate to an immunoglobulin antibody, in particular to a method for increasing the sialic acid content on an immunoglobulin antibody by catalyzing a glycosyltransferase and its application. Provided are a preparation method and application of an immunoglobulin antibody with high sialic acid content. Cytidine-5'-phosphate-sialic acid synthase catalyzes the reaction of sialic acid with cytidine-5'-triphosphate to generate cytidine-5'-phosphate-sialic acid; β1,4-galactosyltransferase converts uridine diphosphate -Galactose as a substrate, catalyzes the connection of galactose to the terminal N-acetylglucosamine of immunoglobulin sugars; sialyltransferase catalyzes the connection of sialic acid to cytidine-5'-phosphate-sialic acid On the galactose at the glycosyl-terminus of immunoglobulins. The prepared immunoglobulin antibody with high sialic acid content can be used to prepare medicines for treating autoimmune diseases, especially rheumatoid arthritis.

Description

technical field [0001] The invention relates to an immunoglobulin antibody, in particular to a method for catalyzing the increase of sialic acid content on the immunoglobulin antibody by using glycosyltransferase and its application. Background technique [0002] Immunoglobulin antibodies, such as intravenous immunoglobulin G, are widely used in the treatment of chronic autoimmune diseases, etc. (1. Imbach, P., Barandun, S., d'Apuzzo, V., et al, High-dose intravenous gammaglobulin foridiopathic thrombocytopenic purpura in childhood, Lancet i, 1228 (1981); 2. Blanchette, V, Imbach, P., Andrew, M., et.al. Randomised trial of intravenous immunoglobulin G, intravenous anti-D, andoral prednisone in Childhood acute immune thrombocytopenic purpura, Lancet 344, 703(1994); 3.Y.Sultan, M.D.Kazatchkine, P.Maisonneuve, U.E.Nydegger, Anti-idiotypic suppression of autoantibodies to factor VIII(antihaemophilic factor, Lagtravenous factor) by high-dose in 765(1984); 4.D.R.Jayne, M.J.Davies...

Claims

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Application Information

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IPC IPC(8): A61P29/00A61P19/02C12P21/02A61P37/02A61K39/395
Inventor 韩守法韩家淮
Owner XIAMEN UNIV
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