Cells producing antibody compositions

A technology of antibody composition and cells, applied in the direction of fusion cells, antibody mimics/scaffolds, antibodies, etc., can solve problems such as determining effector functions

Inactive Publication Date: 2012-01-11
KYOWA HAKKO KIRIN CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in reality, the structures of sugar chains are diverse and complex, and it cannot be said that the structures actually important for effector functions are determined

Method used

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  • Cells producing antibody compositions
  • Cells producing antibody compositions
  • Cells producing antibody compositions

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0614] Generation of anti-ganglioside GD3 human chimeric antibody:

[0615] 1. Construct a tandem expression vector pChiLHGM4 for anti-ganglioside GD3 human chimeric antibody

[0616] Using a DNA ligation kit (manufactured by Takara Shuzo), the plasmid pChi641LGM40 was constructed by ligating an approximately 4.03 kb fragment containing the L-chain cDNA and an approximately 3.40 kb fragment containing the G418 resistance gene and splicing signal, the former by using the restriction enzyme MluI (Takara Shuzo manufactured) and SalI (manufactured by Takara Shuzo) digested pChi641LGM4 of the L chain expression vector, anti-ganglioside GD3 human chimeric antibody (hereinafter referred to as "anti-GD3 chimeric antibody") [J.Immunol.Methods, 167 , 271 (1994)], the latter was obtained by digesting the animal cell expression vector pAGE107 using restriction enzymes MluI (manufactured by Takara Shuzo) and SalI (manufactured by Takara Shuzo) [Cytotechnology, 3, 133 (1990)], and then tra...

Embodiment 2

[0646] Activity assessment of anti-GD3 chimeric antibody:

[0647] 1. Binding activity of anti-GD3 chimeric antibody to GD3 (ELISA)

[0648] The binding activity of the five purified anti-GD3 chimeric antibodies to GD3 (manufactured by Snow Brand Milk Products) in Item 4 of Example 1 was determined by the ELISA method shown in Item 3 in Example 1. figure 2 The detection results of the binding activity determined by changing the concentration of the added anti-GD3 chimeric antibody are shown. as in figure 2 As shown in , five anti-GD3 chimeric antibodies showed almost the same binding activity as GD3. This result shows that the antigen-binding activity of these antibodies is invariant independent of the antibody-producing cells and culture method. It was also shown from the comparison of the NSO-GD3 chimeric antibody (302) with the NSO-GD3 chimeric antibody (GIT) that the antigen-binding activity was unchanged regardless of the medium used in the culture.

[0649] 2. In v...

Embodiment 3

[0660] Preparation of anti-human interleukin-5 receptor alpha chain human CDR-grafted antibody:

[0661] 1. Preparation of cells that stably produce anti-human interleukin-5 receptor α chain human CDR-grafted antibody

[0662] (1) Rat myeloma YB2 / 0 cells were used to prepare antibody-producing cells

[0663] Using the anti-human interleukin 5 receptor α chain human CDR-grafted antibody (hereinafter referred to as "anti-hIL-5Rα CDR-grafted antibody") expression vector, pKANTEX1259HV3LV0 described in WO97 / 10354, was prepared as follows capable of stably producing anti-hIL-5Rα CDR Cells transplanted with antibodies.

[0664] After transplanting 5 μg of anti-hIL-5Rα CDR into the antibody expression vector pKANTEX1259HV3LV0 by electroporation [Cytotechnology, 3, 133 (1990)] introducing 4×10 6 After inoculation into rat myeloma YB2 / 0 cells, the cells were suspended in 40 ml of RPMI640-FBS (10), and distributed to 96-well plates (manufactured by Sumitomo Bakelite) at 200 µl / well. ...

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Abstract

Cells to be used in producing antibody compositions, for example, an antibody having a high antibody-dependent cellular cytotoxic activity which is useful in various diseases, an antibody fragment or a fused protein having the Fc domain of the antibody; a process for producing an antibody composition by using these cells; antibody compositions; and uses thereof. In the above-described antibody compositions, the ratio of sugar chains, which are free from fucose bonded to N-acetylglucosamine at the sugar chain reducing end, to the total N-glycoside linkage complex sugar chains bonded to the Fc domain amounts to 20% or more. Moreover, novel GDP-mannose 4,6-dehydrogenase, GDP-keto-6-deoxymannose 3,5-epimerase 4-reductase, GDP-beta-L-fucose pyrophosphorylase, alpha-1,6-fucosyltransferase and DNAs encoding the same are provided.

Description

[0001] This application is a divisional application of the application number 01819524.5 submitted on October 5, 2001, and the title of the invention is "Cells for Producing Antibody Composition". technical field [0002] The present invention relates to a cell capable of producing antibody molecules, such as antibodies for various diseases, fragments of antibodies, fusion proteins or analogs with Fc regions of antibodies, and processes for producing antibody compositions using the cells, antibody Compositions and uses thereof. Background technique [0003] Since antibodies have high binding activity, binding specificity, and high stability in blood, attempts have been made to use antibodies to diagnose, prevent, and treat various human diseases [Monoclonal Antibodies: Principles and Applications] , Wiley-Liss, Inc., Chapter 2.1 (1995)]. In addition, attempts have been made to produce humanized antibodies, such as human chimeric antibodies or antibodies to which human compl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/00C07K16/00A61K39/395A61P35/00A61P37/00A61P37/02A61P29/00A61P9/00A61P31/04A61P31/12C07K16/06C07K16/18C07K16/28C07K16/32C12N5/10C12N9/04C12N9/10C12N9/12C12N9/88C12N9/90C12P21/08
CPCC12N9/1241C07K16/32C07K16/2866C12N9/1051C07K16/3084C12N9/0006C07K16/00C12P21/005C07K16/065C07K2317/732C07K2317/24C07K2317/41C12N9/90A61K2039/505C07K16/18C07K2319/30A61P29/00A61P31/00A61P31/04A61P31/12A61P35/00A61P37/00A61P37/02A61P37/06A61P37/08A61P9/00A61P9/12C12N5/16
Inventor 神田丰佐藤光男中村和靖内田和久新川丰英山根尚子保坂绘美山野和也山崎基生花井陈雄
Owner KYOWA HAKKO KIRIN CO LTD
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