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Treatment of thrombocytopenia

A technology for thrombocytopenia and hemoglobin, applied in blood diseases, anti-blood group antigen immunoglobulin, extracellular fluid diseases, etc., can solve ineffectiveness, prolonged half-life of red blood cells, monoclonal anti-D antibody can not be used to treat autoimmunity, etc. problem, to achieve the effect of effective ITP treatment and safe ITP treatment

Inactive Publication Date: 2011-09-14
SYMPHOGEN AS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Thus, administration of monoclonal anti-D antibodies results in increased erythrocyte half-life compared to polyclonal anti-D products (Kumpel 1995; Kumpel 2003; Miescher 2004)
Reports suggest that monoclonal anti-D antibodies exhibit no efficacy in ITP patients (summarized in Scaradavou et al., 1997) and at least one report (Godeau et al., 1997, Treatment of chronic autoimmune thrombocytopenic purpura with monoclonal anti-D, Transfusion, 36 :328-30) demonstrated that monoclonal anti-D antibodies cause hemolysis and even anemia, thus inferring that this monoclonal anti-D antibody cannot be used to treat autoimmune ITP

Method used

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  • Treatment of thrombocytopenia
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0197] Production of recombinant polyclonal antibodies against Rhesus D

[0198] donor

[0199] Donors were registered with Aalborg Sygehus Nord. A total of 8 RhD(-) females were immunized with RhD(+) erythrocytes derived from RhD(+) individuals. The donors had different immunization histories with respect to the number of boosts and the source of RhD(+) red blood cells used for immunization. Immunization histories of different donors are given in Table 1.

[0200] Table 1

[0201] Donor ID#

Boost times

Number of promotions from different sources

1

3

2

2

6

2

[0202] 3

2

1

4

4

4

5

2

2

6

3

2

7

2

2

8

2

2

[0203]Monocytes were collected by leukapheresis 5-7 days after the last boost. Cells were pelleted from a commercially available RNA preparation kit and immediately transferred to cell lysis solut...

Embodiment 2

[0288] Generation of polyclonal cell pools for larger scale production

[0289] 27 cell cultures were selected to constitute polyclonal cell lines (RhD157.119D11, RhD159.119B09, RhD160.119C07, RhD161.119E09, RhD162.119G12, RhD163.119A02, RhD189.181E07, RhD191.1199G08, RhD192.1199E08, RhD192. 126H11、RhD197.127A08、RhD199.164E03、RhD201.164H12、RhD202.158E07、RhD203.179F07、RhD207.127A11、RhD240.125A09、RhD241.119B05、RhD244.158B10、RhD245.164E06、RhD293.109A09、RhD301.160A04、 RhD305.181E06, RhD306.223E11, RhD307.230E11, RhD319.187A11 and RhD324.231F07).

[0290] In addition to the high degree of diversity among individual clones, clone selection is also based on the growth and production characteristics of individual cell cultures.

[0291] Selection criteria at the cell culture level include:

[0292] I: Doubling time; must be between 24 and 32 hours

[0293] II: Intracellular staining; a homogeneous population of cells must be displayed

[0294] III: Productivity; must exceed 1.5 pg...

Embodiment 3

[0297] This example demonstrates a 25 specific member anti-RhD recombinant polyclonal antibody (rpAb) and plasma derived anti-D product for phagocytosis Baxter displayed comparable biological activity, whereas anti-RhD rpAbs displayed poor antibody-dependent cellular cytotoxicity (ADCC).

[0298] Red Blood Cell Preparation - Freezing

[0299] Whole blood red blood cells (RBCs) were obtained from healthy donors after informed consent from Blood Bank, Aalborg Hospital, DK, frozen by high glycerol technique (38%) and stored at -80°C. Red blood cells were thawed in 12% NaCl (Merck) and citrate-mannitol (LAB20910.0500, Bie & Berntsen) was added after 3 min. Cells were washed 3 times with PBS (Invitrogen, CA, US) and stored at 4°C in 3% ID-Cellstab (DiaMed, Switzerland) solution.

[0300] Preparation of PBMCs

[0301] Leukocyte-containing blood lines from healthy donors were obtained from Blood Bank, National Hospital, Copenhagen, Denmark, and peripheral blood mononuclear ce...

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PUM

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Abstract

The present invention relates to treatment of thrombocytopenia with apharmaceutical composition comprisingrecombinant polyclonal anti-RhesusD antibody productas the active ingredient.

Description

[0001] All patent and non-patent references cited in this application are hereby incorporated by reference in their entirety. technical field [0002] The present invention relates to pharmaceutical and diagnostic compositions comprising anti-RhD recombinant polyclonal antibody products and their use in the treatment of thrombocytopenia. Treatment of thrombocytopenia can be symptomatic, ameliorative, prophylactic and / or curative. Anti-RhD recombinant polyclonal antibody products and their preparation are disclosed in PCT / DK2005 / 000501. Background technique [0003] Rhesus blood group antigens are located on transmembrane erythrocyte proteins, which encompass the so-called C, c, E, e and D antigens. Due to genetic polymorphism, about 16% of the Caucasian population is Rhesus D-negative (RhD(-)). In addition, there are several genetic and serological variants of RhD (divided into classes II-VII), of which RhD VI most clinically relevant. RhD VI (+) individuals can form al...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/00C07K16/34A61K39/395A61P7/00
CPCC07K16/005C07K2317/77A61K2039/507C07K2317/732C07K2317/55C07K2317/56C07K16/34A61P7/00A61P7/04A61P7/08C12N15/11A61K39/395
Inventor 克里斯琴·迈耶安妮·M·V·詹森埃瓦·林登斯特罗姆安·V·埃斯-约翰逊
Owner SYMPHOGEN AS
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