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Multiplexed Fluorometric ImmunoAssay Testing Methodology and Troubleshooting

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Published on: December 12, 2011

Fluorescence linked immunosorbant assays using microtiter plates.

N Velappan¹, J Clements, C Kiss

¹B division, Los Alamos National Laboratory, Los Alamos, New Mexico 87545, United States.

Journal of Immunological Methods

|June 3, 2008

Summary

This summary is machine-generated.

Fluorescent immunosorbant assays (FLISA) offer an effective alternative to traditional enzyme-linked immunosorbent assays (ELISA) for antibody detection. This study optimizes FLISA parameters for reliable plate-based screening, including antibody immobilization and dye selection.

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Area of Science:

Biochemistry
Immunology
Analytical Chemistry

Background:

Fluorescence methods are common for detecting binding events.
Enzyme-linked immunosorbent assays (ELISA) are predominant for microtiter plate screening.
There is a need for alternative, effective plate-based detection methods.

Purpose of the Study:

To explore parameters for effective fluorescence-based detection in microtiter plates.
To establish fluorescent immunosorbant assays (FLISA) as a viable alternative to ELISA.
To optimize FLISA for screening applications, including phage antibody selection.

Main Methods:

Investigated various microtiter plates suitable for fluorescence detection.
Evaluated different immobilization strategies for antibodies and binding targets.
Assessed the performance of diverse fluorescent dyes and fluorescent proteins.

Main Results:

Identified optimal microtiter plates and immobilization techniques for FLISA.
Demonstrated that FLISA can achieve comparable sensitivity and reliability to ELISA.
Successfully applied FLISA in screening phage antibody selections.

Conclusions:

Fluorescent immunosorbant assays (FLISA) are a robust and effective method for plate-based screening.
FLISA provides a valuable alternative to ELISA, particularly for antibody detection and selection.
Optimization of parameters ensures FLISA's utility in various immunological assays.