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Methods for improving antibody production

A technology for antibodies and humanized antibodies, applied in chemical instruments and methods, biochemical equipment and methods, peptides, etc., can solve problems such as Ig secretion defects

Inactive Publication Date: 2009-09-16
IMMUNOGEN INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Proc Natl Acad Sci USA. 1990 Oct;87(20):8135-9; Wiens GD, Lekkerkerker A, Veltman I, Rittenberg MB Mutation of a single conserved residue in VH complementarity-determining region 2 causes severe Ig secretion defect

Method used

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  • Methods for improving antibody production
  • Methods for improving antibody production
  • Methods for improving antibody production

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0265] [129] The taxane preferably used in the preparation of the cytotoxic conjugate is the taxane represented by the structural formula (XI):

[0266]

[0267] [130] Used for

[0268] A method for forming an exemplary taxane that can be used in the cytotoxic conjugator of the present invention, and a method for conjugating a taxane to a cell-binding agent such as an antibody, are described in US Patent Nos. 5,416,064, 5,475,092, 6,340,701, 6,372,738 , 6,436,931, 6,596,757, 6,706,708 and 6,716,821 and U.S. Patent Publication No. 2004-0024049A1 are described in detail.

[0269] CC-1065 analog

[0270] [131] The toxic agent used in the cytotoxic conjugate according to the present invention may also be CC-1065 or a derivative thereof.

[0271] [132] CC-1065 is a potential antitumor antibiotic isolated from the culture broth of Streptomyces zelensis. The in vitro efficacy of CC-1065 is about 1000 times higher than that of commonly used anticancer drugs such as doxorubicin, methotre...

Embodiment

[0327] material

[0328] [178] Passed standard CsCl 2 Purification technology, prepared pSynC242 and control plasmid products. The QuikChange targeted mutagenesis system (#200518) was obtained from Stratagene. RNeasy mini kit (#74104) was purchased from Qiagen Company. The Superscript First Strand Synthesis System (#11904-418) used for the reverse transcriptase reaction was purchased from GibcoBRL. Obtained Cyber ​​Green real-time PCR Master Mix (#4309155) from APPLY BIOSYSTEM. Fluorescence in Conjugated Streptavidin (#016-010-0841 mg / mL) was from Jackson Immuno Research. A 96-well U-bottom plate (#3077) was purchased from FALCON. EZ-Link Sulfo-NHS-LC-Biotin (#21335) was from Pierce.

[0329] method

[0330] Amino acid substitutions in the framework of huC242HC and LC by site-directed mutagenesis

[0331] Primer design

[0332] [179] The complementary PCR primer pair used for the mutation reaction is 30 bases in length and contains the desired nucleotide substitutions in the prime...

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Abstract

The present invention encompasses manufacturing of antibody variants, such as variant of huC242, or fragments thereof, wherein the variants are manufactured by substituting one or more amino acid residues in a parent antibody. Such substitution(s) is preferably done in a variable region framework sequence of the parent antibody comprising a heavy and a light chain. As a consequence of such substitution(s), variant antibodies or fragments thereof show enhanced antibody synthesis when introduced in a host cell as compared to the parent antibody.

Description

[0001] [01] This application claims priority to U.S. Provisional Application No. 60 / 855,361 filed on October 31, 2006, the entire contents of which are hereby incorporated by reference. Technical field [0002] [02] The present invention relates to methods for increasing antibody production. More specifically, it relates to a method of genetically engineering reengineering antibodies so that the yield of genetically engineered antibodies produced in host cells is greater than that of parental antibodies of genetically engineered antibodies. Background technique [0003] [03] Monoclonal antibodies have a variety of applications, including in vitro diagnostics, laboratory reagents, and therapy. Currently, there are at least 200 kinds of antibodies or antibody fragments undergoing clinical trials (Morrow, K.J., Jr., Monoclonal Antibody Production Technology, Gen. Eng. News, 2002, 20(14): 21). [0004] [04] High-level expression of antibodies in CHO cells requires optimal efficiency f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H21/02C07K16/00C12N5/02
CPCC07H21/02C07K16/00C07K16/2884C07K2317/24
Inventor 周晓迈丹尼尔·塔瓦雷斯
Owner IMMUNOGEN INC