Related Experiment Video
Updated: Nov 11, 2025
Development of a Lateral Flow Immunochromatographic Strip for Rapid and Quantitative Detection of Small Molecule Compounds
Published on: November 13, 2021
A novel antigen immunochromatography fluorometric strip for rapid detection and application of pathogenic bacterial
Shuiqin Fang1, Cheng Liu1, Shaoye Wan2
1School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai 20093, China.
Insights
A novel antigen immune chromatography fluorometric strip (AICFS) offers a faster, more sensitive method for detecting high-affinity antibodies against pathogenic bacteria. This rapid tool aids in screening hybridomas and can be a valuable alternative to traditional ELISA methods.
Area of Science:
- Immunology
- Biotechnology
- Analytical Chemistry
Background:
- Traditional immunoassays often rely on antibodies for detection lines.
- Developing rapid and sensitive methods for high-quality antibody detection is crucial for various applications.
Purpose of the Study:
- To develop and evaluate a novel antigen immune chromatography fluorometric strip (AICFS) for detecting pathogenic bacterial antibodies.
- To compare the performance of AICFS with traditional indirect enzyme-linked immunosorbent assay (ELISA).
Main Methods:
- Development of AICFS using inactivated bacterial antigen as the test line and goat anti-mouse IgG-FITC as the tracer.
- Detection of hybridoma cell cultures for specific bacterial strains (E. coli O157:H7, Acidovorax citrulli, Vibrio parahemolyticus).
- Measurement and comparison of antibody affinity constants (Ka).
Main Results:
- AICFS demonstrated at least 2-fold higher sensitivity than indirect ELISA for detecting E. coli O157:H7 (D3) antibodies.
- AICFS showed high specificity for D3, correctly identifying strains missed by indirect ELISA.
- The method proved effective in various real samples for antibody preparation and screening.
Conclusions:
- AICFS is a sensitive and specific method for detecting high-affinity antibodies against pathogenic bacteria.
- AICFS offers a rapid, time-efficient alternative to indirect ELISA for antibody detection and hybridoma screening.
- This novel approach expands the utility of immune chromatography for antibody applications.
Abstract:
Unlike traditional immunoassay strips, a novel antigen immunechromatography fluorometric strip (AICFS) using inactivated bacterial antigen instead of an antibody as a test line and goat anti-mouse IgG-FITC as a tracer was developed. The applicability survey of AICFS indicated that E. coli O157:H7 (D3) and Acidovorax citrulli (6F) hybridoma cell cultures could be detected, but Vibrio parahemolyticus (H7, C9) hybridoma cell cultures were missed compared with the indirect enzyme-linked immunosorbent assay (ELISA). The four antibody affinity constants (Ka) were measured and compared, and AICFS could be suitable for high-affinity antibody detection. Compared with the traditional indirect ELISA, the AICFS sensitivity for D3 cell cultures, ascites, and purified antibodies was at least 2-fold more sensitive, the AICFS specific for D3 cell cultures by comparative interpretation was compliant except for the strain ATCC 43895, and the indirect ELISA missed it. More importantly, the AICFS method was confirmed by various real samples that it could be used in different scenarios regarding the antibody, including McAb preparation, the effective antibody use, and high-affinity antibody-secreted hybridoma auxiliary preparation and screening. It could be an excellent alternative method with less than 5% corresponding processing time for indirect ELISA method for pathogenic bacterial high-quality antibody detection. This is the first report of using AICFS for bacterial high-quality antibody detection and application in different samples, which demonstrates a rapid auxiliary tool for high-affinity antibody secreted-hybridoma screening and an excellent alternative method for high-quality antibody application.
