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Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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FlowLITE: A protocol to characterize and quantify total antibody isotypes in human plasma using flow cytometry.

Hyunjae J Hahm1, Alexandra F Nazzari2, Benjamin R Babcock1

  • 1Department of Medicine, Division of Immunology, Lowance Center for Human Immunology, Emory University, Atlanta, GA, USA.

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Summary

This study details the Flow Cytometry Light-chain and Isotype Expression (FlowLITE) assay protocol for quantifying human plasma antibody light-chain and isotype expression. The method allows for high-throughput analysis using small sample volumes.

Keywords:
High-hroughput screeningantibodyimmunology

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Area of Science:

  • Immunology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Antibody characterization is crucial for understanding immune responses and disease.
  • Existing methods for antibody isotype and light-chain quantification can be resource-intensive.

Purpose of the Study:

  • To provide a detailed protocol for the Flow Cytometry Light-chain and Isotype Expression (FlowLITE) assay.
  • To enable multiplexed quantification of antibody light-chain and isotype expression in human plasma.

Main Methods:

  • The Flow Cytometry Light-chain and Isotype Expression (FlowLITE) assay utilizes bead-based flow cytometry.
  • The protocol involves antibody titration, multiplexed assay bead preparation, sample staining, and data analysis.
  • Quantification of antibody light chains and isotypes in small fluid volumes is achieved.

Main Results:

  • The FlowLITE assay allows for multiplexed quantification of all antibody isotypes.
  • The protocol details steps for characterizing antibody light-chain and isotype expression in human plasma.
  • The method is applicable to various species and fluid specimens.

Conclusions:

  • The Flow Cytometry Light-chain and Isotype Expression (FlowLITE) assay offers a high-throughput method for antibody analysis.
  • This protocol facilitates detailed characterization of antibody expression profiles.
  • The assay is valuable for immunological research and diagnostics.