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Glycan-interacting compounds and methods of use

A technology of polysaccharides and compositions, which is applied in the field of polysaccharide interaction compounds and uses, and can solve problems such as inability to synthesize

Active Publication Date: 2018-08-28
SEATTLE GENETICS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the loss of functional genes, humans can only synthesize sialic acid in the form of Neu5Ac, but not Neu5Gc

Method used

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  • Glycan-interacting compounds and methods of use
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  • Glycan-interacting compounds and methods of use

Examples

Experimental program
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preparation example Construction

[0174] The preparation of antibodies, whether monoclonal or polyclonal, is known in the art. Techniques for producing antibodies are well known in the art, and are described in, for example, "Antibodies, A Laboratory Manual" by Harlow and Lane, Cold Spring Harbor Laboratory Press, 1988 and "Using Antibodies: A Laboratory Manual" by Harlow and Lane, Cold Spring Harbor Laboratory Press, 1999.

[0175] target

[0176] The glycan-interacting antibodies of the present invention can exert their effects by binding (reversibly or irreversibly) to one or more glycans or glycan-bound or glycan-related targets. In some embodiments, glycan-interacting antibodies can be prepared from any target region taught herein. In some embodiments, the targets of the present invention include glycans. The glycans used to produce antibodies may include at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, At least 12, at least 13, at le...

Embodiment 1

[0731] Example 1. Glycan array analysis

[0732] The optimized glycan array is used to test the affinity and specificity of multiple glycans in a single experiment. The glycan array includes 71 chemically synthesized and clearly defined glycans, most of which are Neu5Ac and Neu5Gc glycan pairs. Array slides are commercially available (ArrayIt Corp, Sunnyvale, CA) and include the glycans listed in the table below.

[0733] Table 12. Array Glycan

[0734]

[0735]

[0736]

[0737] 300ml epoxy blocking buffer was prepared by combining 15ml 2M Tris buffer (pH8) with 0.9ml 16.6M ethanolamine and 284.1ml distilled water. The solution was brought to a final pH of 9.0 with HCl. The solution was filtered using a 0.2 μM nitrocellulose membrane. Preheat the epoxy buffer solution and 1 liter of distilled water to 50°C. The slide is placed on a slide holder and quickly immersed in a staining bucket with heated epoxy blocking buffer. The slides were incubated in epoxy blocking buffer at 50...

Embodiment 2

[0739] Example 2. Antibody binding analysis based on flow cytometry

[0740] Analysis based on flow cytometry was performed to clarify the dose-response curve of antibody binding to cell surface antigen. For these analyses, various cell lines were used.

[0741] MDA-MB-231 cells are human breast cancer cells. They are grown in Earle's minimum essential medium supplemented with 10% fetal calf serum (FCS), 100 μg / ml penicillin, 100 UI / ml streptomycin, and 45 μg / ml gentamicin. MCF-7 cells are also human breast cancer cells and grow under the same conditions as MDA-MB-231 cells. Overexpression (α-N-acetyl-neuraminyl-2,3-β-galactosyl-1,3)-N-acetylgalactosamine α-2,6-sialyltransferase I (GalNAcα2,6- Sialyltransferase I or ST6GalNAcI) stably transfected forms MDA-MB-231 (MDA-MB-231-STn, clone TAH3.P10) and MCF-7 cells (MCF-7 cells clone A12.1) are also Cultured under the same conditions, except that 1mg / ml G418 was added to support the cells expressing the transgene. ST6GalNAc I is ...

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Abstract

The present invention provides glycan-interacting antibodies and methods for producing glycan-interacting antibodies useful in the treatment and prevention of human disease, including cancer. Such glycan-interacting antibodies include humanized antibodies, derivatives and fragments thereof as well as related compositions and kits. Methods of using glycan-interacting antibodies for treatment and diagnosis are included.

Description

[0001] Cross references to related applications [0002] This application requires U.S. Provisional Application No. 62 / 254,278, filed on November 12, 2015, entitled Glycan Interaction Compounds and Methods of Use, and filed on January 4, 2016, entitled Glycan Interaction Compounds and Methods of Use U.S. Provisional Application No. 62 / 274,572, filed on January 27, 2016, titled Glycan Interaction Compounds and Methods of Use, U.S. Provisional Application No. 62 / 287,666, filed on February 11, 2016, titled Glycan U.S. Provisional Application No. 62 / 293,989 for Interacting Compounds and Methods of Use, U.S. Provisional Application No. 62 / 345,515, filed on June 3, 2016, entitled Glycan Interaction Compounds and Methods of Use, and on September 2, 2016 The priority of U.S. Provisional Application No. 62 / 382,835, titled Glycan Interacting Compounds and Methods of Use, filed on Japan, each of which is incorporated herein by reference in its entirety. [0003] Sequence Listing [0004] This ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395C07K16/00C07K16/30G01N33/574
CPCC07K16/3092C07K16/3084C07K2317/565C07K2317/56C07K2317/567C07K2317/92A61K47/68031A61K2039/505C07K2317/24C07K2317/33C07K2317/77A61K47/6851A61P35/00A61P43/00C07K16/30C07K16/00G01N33/574A61K39/395G01N2333/4725A61K47/6817A61K2039/572C07K2317/73
Inventor D·A·埃瓦罗恩J·M·普伦德加斯特J·贝伦斯A·A·卢格夫斯科伊D·T·德兰斯菲尔德
Owner SEATTLE GENETICS INC
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