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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 artificial cell constructs, antibodies, animal cells, etc., can solve the problem of lack of difference

Inactive Publication Date: 2013-12-11
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 , vol10Clin 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

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Experimental program
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Embodiment

[0200] Materials and methods

[0201] Sequencing mRNA from hybridomas

[0202] RNA was prepared using the Qiagen RNeasy Mini kit (cat# 74104). On day 4, 13 ml of the 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 5 times through a 22g needle, add 600 μl 70% EtOH and mix. Two 700 μl aliquots were applied to the RNeasy column, centrifuged for 30 sec and washed with 700 μl RW1. Wash twice with 500 μl PPE, dry for 1 min, and elute twice with 50 μl water.

[0203] 2 μl of RNA was reverse transcribed with the Promega Reverse Transcription System (Catalog # A3500) using oligo dT primers. Reactions were 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. A 1 μl aliquot of cDNA was subjected to PCR with primers selected from one of the N-terminus or 5'-terminus and one o...

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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] This application is a divisional application of Chinese patent application No. 200680048006.X, and the parent application is a Chinese national phase application of PCT / US2006 / 041656 filed on October 23, 2006. [0002] related application [0003] This application claims priority under 35 U.S.C. §119 to U.S. Provisional Application Serial No. 60 / 729,054, filed October 21, 2005. The entire contents of which are incorporated herein by reference. [0004] governmental support [0005] Some aspects of this application may have been made using grants from the National Cancer Institute SBIR Grant No. 5R43CA107608-02. Accordingly, the government may have certain rights in this invention. technical field [0006] The present invention relates to antibodies having enhanced antibody-dependent cellular cytotoxicity (ADCC) activity, methods for their production and methods for their use. Background technique [0007] In the arsenal of disease treatment, monoclonal antibodies ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/00C07K16/04G01N33/577A61K39/395A61P35/00A61P37/00A61P37/02C12N5/071A01K67/027
CPCC07K16/2878C07K16/283C07K2317/12C07K2317/734C07K16/04C07K2317/732A61K2039/505C07K2317/72C07K2317/41C07K16/2875C07K2317/24C07K2317/52C07K2317/71A61P1/04A61P19/02A61P29/00A61P35/00A61P35/02A61P37/00A61P37/02A61P37/06A61P43/00A61K39/395C07K16/18
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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