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Antibody and functional fragment thereof

A technology of functional fragments and antibodies, applied in the direction of antibodies, anti-tumor drugs, radioactive preparations in vivo, etc., can solve problems such as high recurrence rate, no characteristic symptoms, and difficult cancer treatment

Pending Publication Date: 2021-10-15
SUMITOMO CHEM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in many cases, there are no characteristic symptoms in the early stages, and by the time the cancer is diagnosed, the cancer has progressed and treatment has become difficult
In addition, for cancers such as pancreatic cancer, even if the cancer tissue can be surgically removed, the recurrence rate after surgery is high, and postoperative monitoring is essential

Method used

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  • Antibody and functional fragment thereof
  • Antibody and functional fragment thereof
  • Antibody and functional fragment thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0142] Example 1. Preparation of mouse monoclonal antibody against human ES cells

[0143] (a) Preparation of hybridomas

[0144] In DMEM / F12 medium (manufactured by Sigma-Aldrich, catalog number: D9785 ) containing additives (20% KSR (KnockOut Serum Replacement, manufactured by Invitrogen, catalog number: 10828028), 2 mM glutamine, 0.1 mM non-essential amino acid) Human ES cells (clone name: KhES-1) were cultured on mitomycin-treated primary mouse fibroblasts to maintain undifferentiated potential. Contain enzyme solution (0.25% trypsin, 1mg / ml collagenase IV, 20% KSR, 1mM CaCl 2 ) in PBS (phosphate-buffered saline) was added to undifferentiated human ES cells, and the cells were detached by pipetting and centrifuged (1000 rpm, 3 minutes). PBS was added to undifferentiated human ES cells as a pellet to suspend the cells, then ADJUBANT COMPLETE FREUND (manufactured by DIFCO, catalog number: 263810) was added twice as much, and the mixture was phacoemulsified to prepare a...

Embodiment 2

[0168] Example 2. Binding of 3B1E2 antibody to 3'-sialyllactose

[0169] 100 µl of a 10 µg / ml 3'-sialyllactose-PAA (polyacrylamide) (manufactured by GlycoTech Corporation, catalog number 08-038) in PBS was added to the plate of a MaxiSorp 96-well plate (manufactured by Thermo Fisher Scientific, catalog number: 439454). per well, and allow the plate to sit at room temperature for 16 hours. Next, each well was washed 3 times with 300 μl of PBS containing 1% Triton-X100 (wash solution), then 200 μl of PBS containing 5% BSA and 1% Triton-X 100 was added to each well, and the plate was allowed to stand at room temperature 1 hour. Wash each well with 300 µl of washing solution, and then add 100 µl of 3B1E2 antibody (mouse IgG2a or mouse IgM)) or control antibody (mouse IgG2a: manufactured by BioLegend, catalog number: 401502, or mouse IgM: manufactured by BioLegend) to each well , catalog number: 401602), and let the plate sit at room temperature for 2 hours.

[0170] Next, eac...

Embodiment 3

[0172] Example 3. Pathological examination of cancer tissue by 3B1E2 antibody

[0173] A tissue array slide (manufactured by USBiomax, catalog number: MC5003c) on which cancer tissue sections (approximately 15 samples of each type) were placed was deparaffinized (xylene 7 minutes 3 times, methanol 2 minutes 3 times, water), and then add methanol containing 3% hydrogen peroxide to inactivate endogenous peroxidase. After washing with water, the tissue array slides were incubated at 98° C. for 5 minutes in 10 mM citrate buffer (pH 6.0) to activate the antigen. After washing, each sample was surrounded with a Pap pen, and then a Block Ace (manufactured by DS Pharma Biomedical Co., Ltd., catalog number: UKB40) solution was added thereto and subjected to blocking treatment at room temperature for 15 minutes.

[0174] Next, 300 µl of 50 µg / ml 3B1E2 antibody (mouse IgG2a) was added, and the slide was allowed to stand at room temperature for 2 hours. After washing with PBS 3 times,...

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Abstract

Disclosed are: an antibody, which binds to 3'-sialyllactose and contains a heavy chain variable region having a CDR sequence comprising an amino acid sequence ARKNGGLDYAMDY (SEQ ID NO: 3) in which not more than 3 amino acids may be substituted, or a functional fragment thereof; a polynucleotide encoding the antibody or a functional fragment thereof; an expression vector containing the polynucleotide; and a test agent and pharmaceutical composition for treating a disease, which comprise the antibody or a functional fragment thereof.

Description

technical field [0001] The present invention relates to antibodies and functional fragments thereof. In addition, the present invention also relates to a polynucleotide encoding the antibody or its functional fragment, an expression vector containing the polynucleotide, a drug for disease detection and a pharmaceutical composition containing the antibody or its functional fragment. Background technique [0002] Cancer is the leading cause of death in Japan, and the number of cancer deaths is increasing year by year as the average life expectancy increases. Early diagnosis is considered the most important in reducing the number of cancer deaths. In many cases, however, there are no characteristic symptoms in the early stages, and by the time the cancer is diagnosed, the cancer has progressed and treatment has become difficult. In addition, for cancers such as pancreatic cancer, even if the cancer tissue can be surgically removed, the recurrence rate after surgery is high, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/32C12P21/08C12N15/13C12N15/63G01N33/574
CPCC07K16/32G01N33/57407G01N33/57438G01N33/66G01N2440/38G01N2400/38A61P35/00C07K16/44G01N33/574C07K2317/24A61K2039/505A61K51/1045G01N33/57492
Inventor 东清史齐藤幸一
Owner SUMITOMO CHEM CO LTD
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