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Antibodies with enhanced antibody-dependent cellular cytoxicity activity, methods of their production and use

An antibody-dependent, cytotoxic technology, applied in the direction of antibodies, chemical instruments and methods, anti-inflammatory agents, etc., can solve problems such as lack of differences

Inactive Publication Date: 2009-07-15
LFB USA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition to being more active, low-fucose IgG1 is independent of FcγRIII polymorphisms and thus lacks the differences seen with trastuzumab or rituximab in wild-type cells (Niwa et al., 2004b , vol 10 Clin Canc Res)

Method used

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  • Antibodies with enhanced antibody-dependent cellular cytoxicity activity, methods of their production and use
  • Antibodies with enhanced antibody-dependent cellular cytoxicity activity, methods of their production and use
  • Antibodies with enhanced antibody-dependent cellular cytoxicity activity, methods of their production and use

Examples

Experimental program
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Effect test

Embodiment

[0199] Materials and methods

[0200] Sequencing mRNA from hybridomas

[0201] RNA was prepared using Qiagen RNeasy Mini kit (catalog number 74104). On day 4, the 13 ml culture was centrifuged for 5 minutes and resuspended in PBS. Centrifuge again for 5 minutes. The resulting pellet was resuspended in 600 μl RNeasy RLT containing 6 μl μ-ME. Pass the lysate through a 22g needle 5 times, add 600 μl of 70% EtOH and mix. Add 700 μl aliquots to the RNeasy column twice, centrifuge for 30 seconds, and wash with 700 μl RW1. Wash twice with 500 μl PPE, dry for 1 minute, and eluted twice with 50 μl water.

[0202] Using oligo dT primers, 2 μl of RNA was reverse transcribed using Promega Reverse Transcription System (Cat. No. A3500). The reaction was incubated at 42°C for 1 hour, and then heated to 95°C for 5 minutes. The reaction was then diluted to 100 μl with water. PCR was performed on a 1 µl cDNA aliquot with primers selected from one of the N-terminal or 5'-terminal and one of the C-te...

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Abstract

The invention relates, in part, to antibodies with increased ADCC activity. Methods of producing such antibodies are also provided. The antibodies of the invention are produced in mammary epithelial cells, such as those in a non-human transgenic animal engineered to express and secrete the antibody in its milk. The antibodies or compositions comprising the antibodies can be used to treat disease in which ADCC activity provides a benefit. In one embodiment, therefore, the antibodies or compositions comprising the antibodies can be used to treat cancer, lymphoproliferative disease or autoimmune disease.

Description

[0001] Related application [0002] This application claims the priority of U.S. Provisional Application Serial No. 60 / 729054 filed on October 21, 2005 under 35 U.S.C. §119. The entire contents are incorporated herein by reference. [0003] governmental support [0004] Some aspects of this application may be made using funds from the National Cancer Institute SBIR Fund No. 5R43CA107608-02. Therefore, the government may have certain rights in this invention. Technical field [0005] The present invention relates to an antibody with enhanced antibody-dependent cellular cytotoxicity (ADCC) activity, its production method and its use method. Background technique [0006] In the arsenal of disease treatment, monoclonal antibodies are an important weapon. For example, ADCC is a major contributor to the effectiveness of anti-cancer antibodies. The key receptor that mediates ADCC is the FcγIIIa (CD16) receptor, a low-affinity receptor for IgG expressed on natural killer (NK) cells. Mice ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/04A61K39/395C07K16/00
CPCC07K2317/24C07K2317/12C07K16/2875C07K16/04C07K2317/732C07K2317/734A61K2039/505C07K16/283C07K2317/41C07K2317/72C07K16/2878C07K2317/52C07K2317/71A61P1/04A61P19/02A61P29/00A61P35/00A61P35/02A61P37/00A61P37/02A61P37/06A61P43/00A61K39/395C07K16/18
Inventor 丹尼尔·申德勒哈里·M·米德蒂莫西·埃德蒙兹约翰·麦克弗森
Owner LFB USA
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