Preparation of anti-recombinant soluble tissue factor monoclonal antibody by trace injecting antigen into spleen

Xu Cai, Zuo Chun Peng, De Sheng Kong, Hong Shen Guo, Hao Bai, Hou Yan Song, Duan Ma

Research output: Contribution to journalArticlepeer-review

Abstract

Purpose: To prepare the anti-recombinant soluble tissue factor monoclonal antibody to develop an ELISA method which can be used to detect the tissue factor (TF); and to give a reference for going deep into the research and clinical application of TF. Methods: We used TF to immunize Balb/c mice by injecting TF into the spleen directly. Then the spleen cells were separated and fused with mouse myeloma cells (SP2/0) to obtain the hybridoma. The hybridoma was cloned and got 3 clones named as 1-C3, 2-G9, and 3-H9 which could stably secrete TF, and then they were embedded in mice bodies to induce ascites. TF was purified by protein A affinity chromatograph column and labeled with HRP. At last, normal humans' plasma TF levels were determined by ELISA developed from the antibody. Results: The purified TF identified as IgG1 was a single band about M, 150 000 by SDS-PAGE analysis and Western blot showed that it could specifically recognize TF. Normal humans plasma TF was about (130 ± 80) pg/mL as measured by ELISA. Conclusions: We have prepared monoclonal antibody against TF and developed ELISA assays for TF successfully. The research will be useful for TF purification, measurements and clinical study.

Original languageEnglish (US)
Pages (from-to)92-96
Number of pages5
JournalFudan University Journal of Medical Sciences
Volume33
Issue number1
StatePublished - Jan 2006

Keywords

  • Immunity
  • Monoclonal antibody
  • Recombinant soluble tissue factor

ASJC Scopus subject areas

  • Medicine(all)

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