Production of Humanized Antibody against Human High-affinity IgE Receptor in a Serum-free Culture of CHO Cells, and Purification of the Fab Fragments

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概要

• 論文の詳細を見る

• We describe the preparation of Fab fragments of a humanized anti-human high-affinity IgE receptor (fcεRIα) antibody potentially useful for treatment of IgE-mediated allergic diseases. IgE-binding capacities of sixteen combinations of light and heavy chains of four recombinant anti-FcεRIα antibodies, chimeric CRA2,humanized CRA2,chimeric CRA4,and humanized CRA4,were compared. A combination in which both chains were of humanized CRA2 had the highest activity. Stable transfectant clones of four kinds of host cells expressing recombinant antibodies were established. CHO-K1 cells were the most productive. Serum-free media suitable for culture of the stable CHO-transfectant clones were screened. The concentration of the humanized CRA2,which the most productive clone secreted into the chosen serum-free medium, was approximately 100μg/ml. A procedure for the purification of the antibody, papain-digestion, and purification of Fab fragments was established. The highly purified humanized Fab fragments are suitable for use to examine their in vivo activity and immunogenicity in primates.

• 社団法人日本農芸化学会の論文
• 2001-05-23

著者

• 奥村 克純

  三重大院・生資・分子細胞生物学

• OKUMURA KO

  Department of Urology, School of Medicine, Jutendo University

• Ra C

  Atopy (allergy) Research Center Juntendo University School Of Medicine

• Takahashi K

  School Of Life Sciences Tokyo Univ. Of Pharmacy And Life Sciences

• Yokota T

  Foods & Pharmaceuticals Research & Development Laboratory Asahi Breweries Ltd.

• RA Chisei

  Department of Molecular Cell Immunology and Allergology, Advanced Medical Research Center, School of

• Takahashi Kayoko

  Central Research Lnstitute Meiji Milk Products Co. Ltd.

• Tokura Tomoko

  Atopy (allergy) Research Center Juntendo University School Of Medicine

• Takai T

  Atopy (allergy) Research Center Juntendo University School Of Medicine

• Okumura Ko

  東海大学 医学部免疫学教室

• SHIBUYA Ichiro

  Institute for Production Research and Development, The Nikka Whisky Distilling Co., Ltd.

• Ra Chisei

  Allergy And Atopy Research Center Juntendo University School Of Medicine

• Ra Chisei

  Department Of Immunology Juntendo University School Of Medicine

• TAKAI Toshiro

  Bioscience Research and Development Laboratory, Asahi Breweries, Ltd.

• OKUMURA Yasushi

  Bioscience Research and Development Laboratory, Asahi Breweries, Ltd.

• YOKOTA Toyokazu

  Bioscience Research and Development Laboratory, Asahi Breweries, Ltd.

• TAKAHASHI Kyoko

  Bioscience Research & Development Laboratory, Asahi Breweries, Ltd.

• AKAGAWA-CHIHARA Midori

  Bioscience Research & Development Laboratory, Asahi Breweries, Ltd.

• FUKADA Minako

  Bioscience Research & Development Laboratory, Asahi Breweries, Ltd.

• YUUKI Toshihumi

  Bioscience Research & Development Laboratory, Asahi Breweries, Ltd.

• Okumura Ko

  Atopy (allergy) Research Center Juntendo University School Of Medicine

• Okumura Ko

  山梨大学 医学部免疫学講座

• OKUMURA Yasushi

  Applied Technology Research Laboratory Asahi Breweries Ltd

• Okumura Ko

  Department Of Engineering Physics Chubu University

• Yokota T

  Faculty Of Agriculture Shizuoka University

• Shibuya Ichiro

  Institute For Production Research And Development The Nikka Distilling Co. Ltd.

• Fukada Minako

  Bioscience Research & Development Laboratory Asahi Breweries Ltd.

• Yuuki T

  Asahi Breweries Ltd. Ibaraki Jpn

• Akagawa-chihara Midori

  Bioscience Research And Development Laboratory Asahi Breweries Ltd.

• Takahashi Kyoko

  Department Of Food Science And Nutrition School Of Human Environmental Sciences Mukogawa Women'

• Kageyama T

  Kyoto Univ. Aichi Jpn

• Takai Toshiro

  Atopy (allergy) Research Center

• Yokota Takao

  Iss Science Project Office Institute Of Space And Astronautical Science Japan Aerospace Exploration

• Okumura Ko

  Department Of Dermatology Juntendo University School Of Medicine

• Takahashi Kenji

  Laboratory Of Molecular Biochemistry School Of Life Science Tokyo University Of Pharmacy And Life Sc

• Yokota Toyokazu

  Bioscience Research & Development Laboratory, Asahi Breweries, Ltd.

• Takai Toshiro

  Bioscience Research & Development Laboratory, Asahi Breweries, Ltd.

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