Method for inhibiting macrophage infiltration using monoconal anti-alpha-D-antibodies

A monoclonal antibody, macrophage technology, applied in other fields, can solve the problem of indistinguishable

Inactive Publication Date: 2002-06-05
ICOS CORP
View PDF8 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the absence of specific information on the primary structure, it may not be possible to distinguish between these possible cases

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
  • Method for inhibiting macrophage infiltration using monoconal anti-alpha-D-antibodies
  • Method for inhibiting macrophage infiltration using monoconal anti-alpha-D-antibodies
  • Method for inhibiting macrophage infiltration using monoconal anti-alpha-D-antibodies

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Try to detect canine alpha TM1 homologue of

[0056] In an identification dog alpha TM1 In an attempt of the human homologue, the test was performed against the canine α TM1 Cross-reactivity of the specific monoclonal antibody Ca11.8H2 [Moore et al., supra] on human peripheral blood leukocytes. In the presence of 0.1% sodium azide, cell preparations (usually 1×10 6 cells) were incubated on ice with undiluted hybridoma supernatant or purified mouse IgG negative control antibody (10 μg / ml). Binding of monoclonal antibodies was detected by subsequent incubation with 6 μg / ml FITC-conjugated horse anti-mouse IgG (Vector Laboratories, Burlingame, CA). Stained cells were fixed with 2% w / v paraformaldehyde in phosphate buffered saline (PBS) and analyzed with a Facstar Plus fluorescence activated cell sorter (Becton Dickinson, Mountain View, CA). Typically, 10,000 cells are analyzed with logarithmic amplification of fluorescence intensity.

[0057] The result...

Embodiment 2

[0060] Affinity purified canine alpha TM1 for N-terminal sequencing

[0061] Affinity purified canine alpha TM1 To determine the N-terminal amino acid sequence for the design of oligonucleotide probes / primers. In short, the anti-alpha TM1 Monoclonal antibody Ca11.8H2 with Affigel  10 Chromatography resin (BioRad, Hercules, CA) coupled to separate proteins by specific antibody-protein interactions. Antibodies were conjugated to the resin at a concentration of approximately 5 mg antibody / ml resin following the protocol suggested by BioRad. After the conjugation reaction, excess antibody was removed and the resin was blocked with three volumes of 0.1 M ethanolamine. The resin was then washed with 30 column volumes of phosphate buffered saline (PBS).

[0062] 25 grams from a single dog spleen 23 grams of a single dog spleen was homogenized in 250 ml of 25 mM Tris-HCl pH 8.0 buffer containing 0.32 M sucrose and protease inhibitors. Nucleic acids and cellular ...

Embodiment 3

[0077] Large scale affinity purification of canine alpha TM1 for in-house sequencing

[0078] To provide additional amino acid sequences for primer design, purified canine α TM1 For in-house sequencing. Three frozen spleen sections (about 50 g each) and frozen cells from two partial spleens from adult dogs were used to generate proteins for in-house sequencing. 50 g of spleen was homogenized in 200-300 ml of boric acid buffer with a Waring blender. The homogenized material was diluted with 1 volume of buffer containing 4% NP-40, and the mixture was stirred gently for at least 1 hour. Large debris in the resulting lysate was removed by centrifugation at 200Og for 20 minutes, then filtered through either a Corning (Corning, NY) prefilter or a Corning 0.8 micron filter. The lysate was further clarified by passing through a Corning 0.4 micron filter system.

[0079] Spleen lysate and antibody-conjugated Affigel as described in Example 2  10 Resins were mixed in 1...

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

PropertyMeasurementUnit
absorbanceaaaaaaaaaa
Login to view more

Abstract

Methods of inhibiting inflammation and macrophage infiltration following spinal cord injury and methods of modulating the release of TNF-α from cells expressing α-d are disclosed. Anti-α-d monoclonal antibody was used to block the binding between α-d and VCAM-1. A hybridoma secreting the antibody and a diagnostic method using the antibody are disclosed.

Description

[0001] This application is a continuation-in-part of pending U.S. Patent Application Serial No. 09 / 193,043, filed November 16, 1998, which is a continuation-in-part of pending U.S. Patent Application Serial No. 08 / 943,363, filed October 3, 1997 , U.S. Patent Application Serial No. 08 / 943,363 is a continuation-in-part of U.S. Patent Application Serial No. 08 / 605,672 filed on February 22, 1996 and granted as U.S. Patent 5,817,515 on October 6, 1998, U.S. Patent Application Serial No. 08 / 605,672 is a continuation-in-part of U.S. Patent Application Serial No. 08 / 362,652 filed on December 21, 1994 and granted as a continuation-in-part of U.S. Patent 5,766,850 filed on August 5, 1994 U.S. Patent Application Serial No. 08 / 286,889 filed on November 28, 1995 and granted as a continuation-in-part of U.S. Patent No. 5,470,953, U.S. Patent Application Serial No. 08 / 286,889, which in turn was filed on December 23, 1993, and U.S. Patent Application Serial No. 08 / 173,497 and was granted Augus...

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): C07K16/28C12N5/12G01N33/566G01N33/48A61K38/00A61K39/395A61P1/00A61P3/10A61P9/10A61P11/00A61P11/06A61P11/16A61P19/02A61P25/00A61P31/00C07K14/705C12N5/10C12N15/02C12N15/09C12N15/12C12P21/08C12Q1/68C12R1/91G01N33/53G01N33/577G01N33/68
CPCC07K2319/00C07K16/2845A01K2217/05G01N33/6893A61K38/00C07K14/70553A61K2039/505C12Q1/6818G01N2333/70553A61P1/00A61P3/10A61P9/10A61P11/00A61P11/06A61P11/16A61P19/02A61P25/00A61P31/00
Inventor M·W·加拉廷M·范德维伦
Owner ICOS CORP
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap