Antibody Fragments as Inhibitors of Japanese Radish Acid Phosphatase

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概要

• 論文の詳細を見る

• V_H (heavy-chain variable region) and V_L (light-chain variable region) genes were amplified by PCR from hybridomas producing MAb-11 and MAb-18 which inhibited Japanese radish acid phosphatase. Nucleotide sequencing of the V genes demonstrates that the MAbs contained similar V_H and identical V_L domains. Initially, the V_H and V_L genes were expressed in Escherichia coli as single-chain Fv (ScFv) fragments. Fragments ScFv-11 and ScFv-18,named for MAb-11 and MAb-18,respectively, inhibited the enzyme activity to the same extent as the intact MAbs. Both of the antibody fragments widely cross-reacted with other phosphatases, including some phosphomonoesterases and phosphodiesterases from different sources. ScFv-18 also inhibited acid phosphatase from a different origin, but stimulated the activity of alkaline phosphatase from calf intestine. The PCR-amplified V_H and V_L genes were subsequently expressed separately in Escherichia coli as fusion products with glutathione S-transferase. The fusion proteins had little effect on Japanese radish acid phosphatase. Furthermore, a large number of recombinant ScFv fragments specific to the acid phosphatase were generated by using a bacteriophage expression system and a mouse ScFv gene library. These ScFv fragments had a range of effects on the enzyme activity, including inhibition, stimulation, and none. Among them, an ScFv fragment, designated ScFv-G7,inhibited more strongly than ScFv-11 and ScFv-18.

• 社団法人日本農芸化学会の論文
• 1998-06-23

著者

• Miyamoto Yoshinori

  Department of Cardiology, Kawasaki Medical School

• Sakamoto J

  Shimane Univ. Matsue Jpn

• 宮本 敬久

  Laboratory Of Food Hygienic Chemistry Division Of Food Biotechnology Department Of Bioscience And Bi

• MIYAMOTO Takahisa

  Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School, Kyushu Universi

• HATANO Shoji

  Graduate School of Health and Social Welfare Science, Nishikyushu University

• Honjoh K

  Kyushu Univ. Fukuoka Jpn

• Honjoh Ken-ichi

  Laboratory Of Food Hygienic Chemistry Department Ofbioscience And Biotechnology Faculty Of Agricultu

• Honjoh Ken-ichi

  Department Of Bioscience And Biotechnology Faculty Of Agriculture Graduate School Kyushu University

• Sakamoto J

  Food Research Laboratories Ajinomoto Co. Inc.

• HONJOH Kenichi

  Department of Bioscience and Biotechnology, Division of Bioresource and Bioenvironmental Sciences, G

• TAKATA Ryoji

  Department of Bioscience and Biotechnology, Division of Bioresource and Bioenvironmental Sciences, G

• SOEDA Takahiko

  Food Research Laboratories, Ajinomoto Co. Inc.

• SAKAMOTO Jiro

  Food Research Laboratories, Ajinomoto Co. Inc.

• HATANO Shoji

  Department of Food Science and Technology, Faculty of Agriculture, Kyushu University

• Yoshimoto Makoto

  Department Of Food Science And Technology Faculty Of Agriculture Kyushu University

• Yoshimoto M

  Department Of Upland Farming Research National Agricultural Research Center For Kyushu Okinawa Regio

• Hatano Shoji

  Department Of Food Science And Technology Faculty Of Agriculture 46-09 Kyushu University

• Takata Ryoji

  Department Of Bioscience And Biotechnology Division Of Bioresource And Bioenvironmental Sciences Gra

• Soeda Takahiko

  Food Research Laboratories Ajinomoto Co. Inc.

• Hatano S

  Kyushu Univ. Fukuoka Jpn

• Miyamoto T

  Kyushu Univ. Fukuoka Jpn

• Tamai Rumiko

  Laboratory Of Food Hygienic Chemistry Faculty Of Agriculture Kyushu University

• Yoshimoto M

  Sweetpotato Breeding Research Team National Agricultural Research Center For Kyushu Okinawa Region

• Miyamoto Yoshinori

  Department Of Cardiology Kawasaki Medical School

• SAKAMOTO Jiro

  Department of Physics, Shimane University

• SAKAMOTO Jiro

  Physics Department, Shimane University

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