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Purification of human brain tissue factor.

S Kang1, J Niemetz

  • 1Mount Sinai School of Medicine, New York, NY.

Thrombosis and Haemostasis
|June 16, 1988
PubMed
Summary

Researchers purified human brain tissue factor (Factor III), essential for blood coagulation, achieving 98% homogeneity. This advancement utilized affinity chromatography for efficient isolation of this crucial glycoprotein cofactor.

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Area of Science:

  • Biochemistry
  • Hematology
  • Molecular Biology

Background:

  • Tissue factor (Factor III) is a critical glycoprotein cofactor.
  • It is essential for initiating both extrinsic and intrinsic blood coagulation pathways.
  • Factor VII requires tissue factor for its catalytic activity.

Purpose of the Study:

  • To develop an efficient purification method for human brain tissue factor.
  • To achieve high purity and yield of the tissue factor.
  • To characterize the purified tissue factor.

Main Methods:

  • Purification of human brain tissue factor from acetone-dried brain powder using Triton X-100 extraction.
  • Affinity chromatography employing immobilized anti-Factor VII-agarose beads.
  • Characterization using SDS-PAGE and isoelectric focusing techniques.

Main Results:

  • Achieved a 2,500-fold purification of tissue factor to 98% homogeneity.
  • Obtained an overall yield of 36% for the purified tissue factor.
  • Determined the apparent molecular weight to be 45,000 Da and isoelectric point to be 4.8-5.1.

Conclusions:

  • A robust method for purifying human brain tissue factor was established.
  • The purified tissue factor is highly homogeneous and characterized.
  • This facilitates further research into the role of tissue factor in coagulation.

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