Unlock instant, AI-driven research and patent intelligence for your innovation.

Use of lectins to promote oligomerization of glycoproteins and antigenic molecules

A lectin and antigen technology, applied in the direction of cancer antigen components, peptide/protein components, vertebrate antigen components, etc., can solve problems such as differences in legume lectins

Inactive Publication Date: 2006-05-31
ANTIGENICS
View PDF27 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although they are similar at the level of primary, secondary and tertiary structure, dimeric / tetrameric aggregates of legume lectins exhibit striking differences in their patterns of quaternary binding and monomeric organization

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Use of lectins to promote oligomerization of glycoproteins and antigenic molecules
  • Use of lectins to promote oligomerization of glycoproteins and antigenic molecules
  • Use of lectins to promote oligomerization of glycoproteins and antigenic molecules

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0376] 6. Example 1: Consistently Elevated Levels of CON A in Human GP96-Peptide Complex Batches

[0377] Tissue homogenates (A to D) from 4 independent human kidney tumor samples were prepared and processed by Con A column chromatography. The Con A eluate was separated and half of the material was set aside. The remaining samples were buffer exchanged to PBS (PD-10 column) followed by further purification of 2 samples over split DEAE columns to generate 2 homogenate-matched final products - 1 with no buffer between Con A and DEAE columns One with a buffer exchange step (Bx) between the two columns. Con A concentrations in these isolated gp96-peptide samples were then determined using a sensitive ELISA for the detection of Con A.

[0378] Concanavalin A specific ELISA:

[0379] Materials: Concanavalin A was purchased from Sigma, catalog number C7275. Capture Antibody: Mouse Anti-Con A Cat. No. MAB 158 Maine Biotechnology, Primary Antibody: Rabbit Anti-Con A Cat. No. C740...

Embodiment 2

[0382] 7. Example 2: CON A exists in an oligomerized molecular complex

[0383] Common homogenates from chemically induced murine fibrosarcoma (Meth A) tissue were prepared and processed by Con A column chromatography. The Con A eluate was separated and half of the material was set aside. The remaining samples were buffer exchanged to PBS, followed by further purification of 2 samples over split DEAE columns to generate 2 homogenate-matched final products - 1 with no buffer exchange between Con A and DEAE columns (no Bx) , 1 with buffer exchange step (Bx). The 2 samples and samples of free Con A (5 μg, 50 μg / mL) were fractionated together by SEC on a Superdex200 column (upper, middle and lower, respectively). Collected fractions were analyzed by SDS-PAGE for gp96 (fractions 1 to 8; inset), and individual fractions were evaluated for Con A content by direct ELISA against Con A (fractions 1 to 14; overlay) (for Con A For direct ELISA of A, see Example 1). There was only a sm...

Embodiment 3

[0385] 8. Example 3: Oligomerization Response Correlates with In Vitro and In Vivo Potency

[0386] 8.1. Con A content in human gp96 samples correlates with potency in vitro

[0387] gp96 samples were prepared from 4 independent human kidney tumor samples (A to D) as described above (see figure 1 ), generating 4 pairs of samples that differed only in the inclusion or omission of the buffer exchange step between the Con A and DEAE columns. Using the CD71 system, the con A content of all eight samples was determined (panel A; see also figure 1 ) and in vitro antigen re-presentation (Panel B). In each case, material prepared by a method that included a buffer exchange step (and contained high levels of Con A) compared to samples generated from matched tumor homogenates and prepared by a step that omitted the buffer exchange step Has higher in vitro re-presentation activity.

[0388] CD71 in vitro re-presentation assay:

[0389] Materials: The antigen presenting cell line R...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The present invention relates to the use of lectins or lectin-like molecules to promote the oligomerization of glycoproteins or complexes containing immunological and / or biological activity of glycoproteins. More specifically, the present invention provides compositions comprising a molecular complex of a lectin molecule and an immunologically and / or biologically active molecule. Also provided are methods of preparing such molecular complexes, of using compositions comprising such molecular complexes to prevent and treat diseases, particularly cancers and infectious diseases, and of eliciting an immune response in a subject.

Description

[0001] This application claims the benefit of US Provisional Application Serial No. 60 / 450,721, filed February 28, 2003, which is hereby incorporated by reference in its entirety. [0002] 1. Introduction [0003] The present invention relates to the fields of biotherapy, immunotherapy and stress protein-mediated immune modulation. More specifically, the present invention relates to the use of molecular complexes containing lectins or lectin-like molecules bound to immunologically and / or biologically active molecules to enhance the role of immunologically and / or biologically active molecules in the prevention or treatment of diseases (especially Compositions and methods for prevention and / or treatment in the prevention or treatment of cancer or infectious diseases). 2. Background of the invention [0004] Citation and discussion of references herein should not be construed as an admission that they constitute prior art to the present invention. [0005] 2.1 Immune response ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/00A61K39/00
CPCA61K2039/55511A61K2039/55516A61P35/00A61P37/00A61K39/4614A61K39/4622A61K39/4611A61K39/464412A61K38/16A61K39/385A61K39/0011A61K2300/00
Inventor J·R·扎布雷基S·A·蒙克斯
Owner ANTIGENICS