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High-throughput Flow Cytometry Cell-based Assay to Detect Antibodies to N-Methyl-D-aspartate Receptor or Dopamine-2 Receptor in Human Serum

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[A method to detect gene transfection efficacy by flow cytometry].

Ran-yi Liu¹, Hui-ling Luo, Hai-lin Feng

¹Biotherapy Center, Cancer Center, Sun Yat-sen University, Guangzhou 510060, P. R. China.

AI Zheng = Aizheng = Chinese Journal of Cancer

|November 28, 2002

Summary

Flow cytometry offers a more sensitive, objective, and accurate method for assessing gene transfer and expression efficiency compared to manual counting. This technique provides reliable data, especially for multiple gene transfers.

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Area of Science:

Molecular Biology
Cell Biology
Biotechnology

Context:

Traditional methods for analyzing gene transfer and expression rely on manual counting of reporter gene-expressing cells, which is subjective and prone to inaccuracies.
Assessing gene transfer efficiency is crucial for various applications, including gene therapy and drug development.

Purpose:

To develop and validate a convenient, objective, and accurate method for assessing gene transfer and expression efficacy in mammalian cells.
To compare the performance of flow cytometry with traditional manual counting techniques.

Summary:

Hepatocellular carcinoma (HCC) cells were infected with a recombinant adenovirus expressing green fluorescent protein (AdCMV/GFP) at varying multiplicities of infection (MOIs).
Transfection rates were assessed by manual counting and flow cytometry (FCM). Fluorescent indices (FIs) were analyzed by FCM.
FCM-based transfection rates correlated logarithmically with vector doses, while FIs showed a strong linear correlation (r = 0.9984, P < 0.001).

Impact:

Flow cytometry provides a more sensitive, objective, and accurate assessment of gene transfer and expression efficiency than manual counting.
This method is particularly advantageous for evaluating multiple gene transfers and expression levels within cells.
The findings suggest flow cytometry as a superior tool for quantitative analysis in gene transfer studies.