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Acute leukemia and lymphoblastic lymphoma-specific CD43 epitope and use thereof

A technology for leukemia and lymphoma, applied in the field of CD43 epitope

Active Publication Date: 2012-10-10
锦湖HT株式会社
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, they are ineffective in detecting or eliminating leukemia or lymphoma cells

Method used

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  • Acute leukemia and lymphoblastic lymphoma-specific CD43 epitope and use thereof
  • Acute leukemia and lymphoblastic lymphoma-specific CD43 epitope and use thereof
  • Acute leukemia and lymphoblastic lymphoma-specific CD43 epitope and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] In order to study specific cell surface proteins on thymocytes, human thymocytes were administered to Balb / c mice to produce antibodies against human thymocytes according to the following examples.

[0061] 10 7 Balb / c mice were immunized intraperitoneally with human thymocytes every two weeks for six weeks. The spleen of the Balb / c mouse was removed 3 days after the last administration to prepare a spleen cell suspension. Monoclonal antibodies were produced by fusing human thymocyte-immunized Balb / c splenocytes with 9-azaguanine-resistant SP2 / 0-Ag14 mouse myeloma cells. The cell fusion method refers to the method of Koeler and Milstein (Koeler & Milstein Nature, 1975, 256, 495-497). Make 10 with 50% polyethylene glycol 4000 8 splenocytes and 10 7 fusion of myeloma cells. Wash the cells and resuspend them in DMEM (Dulbeco's modified Eagle's medium) medium containing 20% ​​bovine serum albumin, 100 μM hypoxanthine, 0.44 μM aminopterin and 16 μM deoxythymidine Glyco...

Embodiment 2

[0064] In order to find clones secreting antibodies that recognize specific cell surface antigens on thymocytes among the hybridoma clones generated in Example 1, according to the avidin-biotin complex obtained by combining avidin and biotin (ABC) staining method, using the supernatant of the hybridoma clones produced in Example 1, immunoperoxidase staining was performed on 4 μm thick fresh tissue sections and formalin-fixed, paraffin-embedded tissue sections. Supernatants from monoclonal cells were used as primary antibodies. Paraffin-embedded tissues were treated with normal mouse serum after paraffin removal and kept for 1 hour to prevent non-specific background staining. After addition of primary antibodies, they were left overnight and washed three times with phosphate-buffered saline (PBS). Biotinylated goat anti-mouse immunoglobulin was added as a secondary antibody. Incubate for 1 hour at room temperature and wash three times with PBS. A conjugate of streptavidin an...

Embodiment 3

[0067] To assess the reactivity of EB-1 antibodies according to the developmental stage of thymocytes, flow cytometry was performed. A human thymus removed from a patient during cardiac surgery is finely minced to prepare a single cell suspension. Will 1×10 6 Cells were suspended in 100 μl PBS and dispensed into individual test tubes. Add 100 µl of EB-1 culture supernatant and stir. The solution was allowed to react at 4°C for 30 minutes, centrifuged at 1,500 rpm for 5 minutes, and the pellet was washed twice with PBS to remove unreacted antibody. This pellet was suspended in 50 µl of a solution containing a diluted secondary antibody (FITC-conjugated goat anti-30 mouse Ig, produced by Zymed), and reacted at 4°C for 30 minutes in the dark. After washing twice, the pellet was suspended in 50 μl solution containing phycoerythrin (PE)-conjugated anti-CD8 antibody and allophycocyanin (APC)-conjugated anti-CD4 antibody, and reacted at 4°C in the dark. 30 minutes, then wash twic...

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Abstract

The present invention relates to a CD43 epitope expressed on human acute leukemia and lymphoblastic lymphoma cells and its use. More particularly, the present invention relates to a CD43 epitope expressed on human acute leukemia, lymphoblastic lymphoma cells, but not on mature hematopoietic cells, hematopoietic stem cells and non-hematopoietic cells, and to its diagnostic and therapeutic application on acute leukemia and lymphoblastic lymphoma.

Description

[0001] Cross References to Related Applications [0002] This application claims U.S. Provisional Applications 60 / 679,910 (filed May 11, 2005) and 11 / 312,126 (filed December 20, 2005) filed with the United States Patent and Trademark Office, and filed with the Korean Intellectual Property Office Korean Patent Application No. 2005-0077906 (filed Aug. 24, 2005) is hereby incorporated by reference in its entirety as the basis of priority and benefit of these applications. technical field [0003] The invention relates to a CD43 epitope expressed on human acute leukemia and lymphoblastic lymphoma cells and its application. Specifically, the present invention relates to a CD43 epitope expressed on human acute leukemia and lymphoblastic lymphoma cells, but not expressed on mature hematopoietic cells and hematopoietic stem cells, and also relates to its expression in acute leukemia and lymphoblastic lymphoma cells. Diagnostic and therapeutic uses of lymphoma cells. Background tech...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/705C07K16/28A61K39/395A61P35/00A61P35/02G01N33/53
CPCA61K39/395A61K47/68A61K2039/505C07K7/06C07K16/2896G01N33/532G01N33/57426G01N2333/70596
Inventor 朴圣会郑景天崔银玲朴承杓
Owner 锦湖HT株式会社
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