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  • 标题:Correction of microplate location effects improves performance of the thrombin generation test
  • 本地全文:下载
  • 作者:Yideng Liang ; Samuel A Woodle ; Alexey M Shibeko
  • 期刊名称:Thrombosis Journal
  • 印刷版ISSN:1477-9560
  • 电子版ISSN:1477-9560
  • 出版年度:2013
  • 卷号:11
  • 期号:1
  • 页码:12
  • DOI:10.1186/1477-9560-11-12
  • 语种:English
  • 出版社:BioMed Central
  • 摘要:

    Background

    Microplate-based thrombin generation test (TGT) is widely used as clinical measure of global hemostatic potential and it becomes a useful tool for control of drug potency and quality by drug manufactures. However, the convenience of the microtiter plate technology can be deceiving: microplate assays are prone to location-based variability in different parts of the microtiter plate.

    Methods

    In this report, we evaluated the well-to-well consistency of the TGT variant specifically applied to the quantitative detection of the thrombogenic substances in the immune globulin product. We also studied the utility of previously described microplate layout designs in the TGT experiment.

    Results

    Location of the sample on the microplate (location effect) contributes to the variability of TGT measurements. Use of manual pipetting techniques and applications of the TGT to the evaluation of procoagulant enzymatic substances are especially sensitive. The effects were not sensitive to temperature or choice of microplate reader. Smallest location effects were observed with automated dispenser-based calibrated thrombogram instrument. Even for an automated instrument, the use of calibration curve resulted in up to 30% bias in thrombogenic potency assignment.

    Conclusions

    Use of symmetrical version of the strip-plot layout was demonstrated to help to minimize location artifacts even under the worst-case conditions. Strip-plot layouts are required for quantitative thrombin-generation based bioassays used in the biotechnological field.

  • 关键词:Thrombin generation test; Location effects; Immunoglobulin; Thrombogenicity
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