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MONOCLONAL ANTIBODY AGAINST EBAG9, HYBRIDOMA, AND APPLICATION FOR MONOCLONAL ANTIBODY AGAINST EBAG9

Foreign code F140008005
File No. S2012-1179-C0
Posted date Oct 28, 2014
Country WIPO
International application number 2013JP074452
International publication number WO 2014042167
Date of international filing Sep 11, 2013
Date of international publication Mar 20, 2014
Priority data
  • P2012-199412 (Sep 11, 2012) JP
Title MONOCLONAL ANTIBODY AGAINST EBAG9, HYBRIDOMA, AND APPLICATION FOR MONOCLONAL ANTIBODY AGAINST EBAG9
Abstract A monoclonal antibody against EBAG9, said monoclonal antibody being obtained by a method including: a step in which a polypeptide is administered to a mouse as an immunogen, said polypeptide containing the amino acid sequence set forth in SEQ ID No. 1, and not containing the full-length amino acid sequence of EBAG9 protein; a step in which antibody-producing cells are collected from the mouse; a step in which an antibody-producing cell and a myeloma are fused to produce a hybridoma; a step in which an antibody produced by the hybridoma is brought into contact with a polypeptide containing the amino acid sequence set forth in SEQ ID No. 1, and not containing the full-length amino acid sequence of EBAG9 protein, and then a hybridoma is selected that produces an antibody that binds to the polypeptide containing the amino acid sequence set forth in SEQ ID No. 1, and not containing the full-length amino acid sequence of EBAG9 protein; and a step in which antibodies produced by the selected hybridoma are collected.
Outline of related art and contending technology BACKGROUND ART
Breast cancer, many women over the age of 40, is an increasing trend in Japan. And said breast cancer is, estrogen receptor positive is often, endocrine therapy such as anti-estrogen agent endocrine therapy is the use of the drugs is known. However, in breast cancer, in the course of treatment, drug resistance to endocrine therapy to obtain many (resistance), and therefore a poor prognosis in question. Therefore, resistance to endocrine therapy to obtain breast cancer diagnosed at an early stage whether or not a technique capable of is demanded.
EBAG9 (Estrogen receptor-binding fragment associated gene 9) is, isolated as estrogen response gene, in later studies, involved in sugar chain modification of cancer cells, avoid from the immune system are shown to be involved. In addition, is EBAG9, breast cancer, ovarian cancer, prostate cancer, liver cancer, renal cancer, bladder cancer, testicular tumor, brain tumor, uterine cancer, thyroid cancer, gallbladder cancer, bile duct cancer, pancreatic cancer, colon cancer, gastric cancer, esophageal cancer, lung cancer, throat pharyngeal cancer, squamous cell carcinoma, malignant lymphoma, malignant mesothelioma, Paget's disease has been reported expression, the progression of a cancer, thought to be involved in metastasis.
So far, as an immunogen, using the entire length of the EBAG9, and developed a polyclonal antibody against EBAG9, wherein the polyclonal antibody immunostaining with breast cancer sample analysis is performed according to the method (for example, see non-patent document 1). The analysis, immunostaining of-EBAG9, estrogen receptor can be correlated to immunostaining, as well as CD3 positive tumor cell infiltration into the tissue can be correlated with an decreased T is shown. However, resistance to endocrine therapy of breast cancer such as whether or not to obtain the relationship between the prognosis of breast cancer, not yet been elucidated.
Therefore, resistance to endocrine therapy to obtain breast cancer diagnosed at an early stage whether or not a technique capable of not developed yet, its rapid development strongly demanded.
In addition, EBAG9 is as described above, breast cancer, ovarian cancer, prostate cancer, liver cancer, renal cancer, bladder cancer, testicular tumor, brain tumor, uterine cancer, thyroid cancer, gallbladder cancer, bile duct cancer, pancreatic cancer, colon cancer, gastric cancer, esophageal cancer, lung cancer, throat pharyngeal cancer, squamous cell carcinoma, malignant lymphoma, malignant mesothelioma, Paget's disease have been reported and expression, antibodies to EBAG9 can be effective in the treatment of cancer prevention to what would be considered of, yet such antibodies may be not provided.
Scope of claims (In Japanese)請求の範囲 [請求項1]
 免疫原として、配列番号1に記載のアミノ酸配列を含み、EBAG9タンパク質の全長のアミノ酸配列は含まないポリペプチドをマウスに投与する工程と、
 前記マウスから抗体産生細胞を回収する工程と、
 前記抗体産生細胞とミエローマとを細胞融合し、ハイブリドーマを調製する工程と、
 前記ハイブリドーマが産生する抗体と、配列番号1に記載のアミノ酸配列を含み、EBAG9タンパク質の全長のアミノ酸配列は含まないポリペプチドとを接触させ、前記配列番号1に記載のアミノ酸配列を含み、EBAG9タンパク質の全長のアミノ酸配列は含まないポリペプチドに結合する抗体を産生するハイブリドーマを選択する工程と、
 前記選択されたハイブリドーマが産生する抗体を回収する工程とを含む方法により得られることを特徴とするEBAG9に対するモノクローナル抗体。

[請求項2]
 エピトープが、配列番号13に記載のアミノ酸配列内にあることを特徴とするEBAG9に対するモノクローナル抗体。

[請求項3]
 請求項1から2のいずれかに記載のEBAG9に対するモノクローナル抗体を産生するハイブリドーマであって、受託番号:NITE BP-01406であることを特徴とするハイブリドーマ。

[請求項4]
 被検体由来の試料中におけるEBAG9を請求項1から2のいずれかに記載のEBAG9に対するモノクローナル抗体を用いて測定する工程と、
 前記測定結果を指標として、内分泌療法薬に耐性を有する乳癌か否かを評価する工程とを含むことを特徴とする内分泌療法薬耐性乳癌の予測方法。

[請求項5]
 請求項1から2のいずれかに記載のEBAG9に対するモノクローナル抗体を含むことを特徴とする内分泌療法薬耐性乳癌の予測用診断薬又はキット。

[請求項6]
 請求項1から2のいずれかに記載のEBAG9に対するモノクローナル抗体を含むことを特徴とする腫瘍増殖抑制剤。

[請求項7]
 請求項1から2のいずれかに記載のEBAG9に対するモノクローナル抗体を含むことを特徴とする癌転移抑制剤。

[請求項8]
 請求項6に記載の腫瘍増殖抑制剤、及び請求項7に記載の癌転移抑制剤の少なくともいずれかを含むことを特徴とする医薬。

  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • SAITAMA MEDICAL UNIVERSITY
  • Inventor
  • INOUE, Satoshi
  • IKEDA, Kazuhiro
  • IJICHI, Nobuhiro
  • SHIGEKAWA, Takashi
  • TSUDA, Hitoshi
  • MIYAZAKI, Toshiaki
IPC(International Patent Classification)
Specified countries National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IS JP KE KG KN KP KR KZ LA LC LK LR LS LT LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN TD TG

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