Studies on Purification of Human Factor VIII, HMW and LMW Subunits and on Properties, of Rabbit Antibodies

  • AIHARA Morio
    1st Depart. of Internal Med. Hirosaki University School of Med.
  • SAWADA Yoshihiko
    1st Depart. of Internal Med. Hirosaki University School of Med.
  • SEGAWA Masanobu
    1st Depart. of Internal Med. Hirosaki University School of Med.
  • KIMURA Asano
    1st Depart. of Internal Med. Hirosaki University School of Med.
  • CHIBA Yoichi
    1st Depart. of Internal Med. Hirosaki University School of Med.
  • YOSHIDA Yutaka
    1st Depart. of Internal Med. Hirosaki University School of Med.

Bibliographic Information

Other Title
  • ヒト血漿第VIII因子,HMW及びLMWの分離,精製とそれらの抗血清に関する検討
  • ヒト血漿第8因子,HMW及びLMWの分離,精製とそれらの抗血清に関する検討
  • ヒト ケッショウ ダイ 8 インシ HMW オヨビ LMW ノ ブンリ セイセ

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Description

Factor VIII was purified from normal human plasma by gel filtration. Purification was 2900 fold with the recovery rate of 5%. This highly purified F-VIII was successfully separated into two components by 0.25 M CaCl2. One which was eluted in the void volume (high molecular weight protein, HMW), contained VWF and AGN. The other in later fraction (low molecular weight protein, LMW), showed only AHF activity.<br>Antisera against F-VIII and against HMW were produced in rabbits. Anti F-VIII antiserum had both AHF and VWF neutralizing effects, and formed a single precipitine line against normal cryoprecipitates. Anti HMW antiserum which inhibited only VWF revealed same immunoprecipitabilities as F-VIII antiserum, and it's inhibitory effect was strong as 8 time as Hoechst's antiserum. Antibody against LMW was not able to be detected.<br>Using this anti F-VIII antiserum, AGN was measured by Laurell methods and crossed immuno-electrophoresis. AGN showed very low level (0-16.5%) in 5 patients with von Willebrand's disease, and electrophoretic abnormality by CIEP was seen in acquired von Willebrand's syndrome (0.40). Abnormal electrophoretic mobility was also seen in 2 patients with liver disease (0.30, 0.43) suggesting the presence of functionaly abnormal F-VIII.<br>The fractions of F-VIII by author's methods would be useful to elucidate the pathophysiological mechanism of F-VIII and VWF in these intriguing group of disorders.

Journal

  • Rinsho Ketsueki

    Rinsho Ketsueki 21 (1), 24-32, 1980

    The Japanese Society of Hematology

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