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Related Experiment Videos

Antibody quantitation in seconds using affinity Perfusion Chromatography.

S P Fulton1, M Meys, L Várady

  • 1PerSeptive Biosystems, Inc., University Park at MIT, Cambridge, MA 02139.

Biotechniques
|August 1, 1991
PubMed
Summary
This summary is machine-generated.

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A novel assay technique uses protein A or G with Perfusion Chromatography for rapid antibody quantification. This method achieves results in under a minute, offering high accuracy and a wide dynamic range.

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Chromatography

Background:

  • Antibody assays are crucial in diagnostics and research.
  • Existing methods can be time-consuming and require sample dilution.
  • Need for rapid, automated, and sensitive antibody detection techniques.

Purpose of the Study:

  • To develop an extremely rapid assay technique for antibody quantification.
  • To utilize protein A or G immobilized on Perfusion Chromatography supports.
  • To achieve high throughput and accuracy in antibody measurements.

Main Methods:

  • Immobilizing protein A or protein G onto Perfusion Chromatography support matrices.
  • Direct injection of antibody samples (dilute or concentrated) onto the column.

Related Experiment Videos

  • Quantification of bound antibody via elution and UV absorbance detection.
  • Utilizing unique mass transport characteristics of the support matrices.

    • Main Results:

    • Achieved total assay cycle times of 1 minute or less, with some assays as short as 15 seconds.
    • Demonstrated accurate quantification over a dynamic range of 100,000:1 or greater without sample dilution.
    • Assay system proved to be extremely repeatable and automatable with conventional HPLC systems.
    • Successfully demonstrated antibody assay in a wide variety of sample types.

    Conclusions:

    • The developed assay technique offers a significant advancement in speed and efficiency for antibody quantification.
    • The use of protein A/G on Perfusion Chromatography supports enables rapid, accurate, and automated antibody detection.
    • This technique has broad applicability across various sample types and research/diagnostic settings.