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Colloidal gold-based immunochromatographic assay for detection of ricin
Rong-Hwa Shyu1, Huey-Fen Shyu, Hwan-Wun Liu
1Institute of Preventive Medicine, National Defense Medical Center, Taipei, Taiwan, Republic of China.
Insights
A novel immunochromatographic assay detects ricin rapidly using specific monoclonal antibodies (Mabs). This assay can identify ricin at 50 ng/ml in under 10 minutes, with sensitivity enhanced to 100 pg/ml.
Area of Science:
- Biotechnology
- Immunology
- Analytical Chemistry
Background:
- Ricin is a highly toxic protein requiring sensitive detection methods.
- Existing ricin detection assays may lack speed or sensitivity.
- Development of rapid, sensitive diagnostic tools is crucial for public health and security.
Purpose of the Study:
- To develop a rapid immunochromatographic assay for ricin detection.
- To utilize monoclonal antibodies (Mabs) for specific ricin detection.
- To achieve high sensitivity and rapid results for ricin identification.
Main Methods:
- A sandwich immunochromatographic assay format was employed.
- Monoclonal antibodies targeting distinct ricin chains (A and B) were used.
- Colloidal gold-conjugated Mabs and immobilized Mabs on nitrocellulose membranes facilitated detection.
Main Results:
- The assay detected ricin at concentrations as low as 50 ng/ml in under 10 minutes.
- Sensitivity was significantly enhanced to 100 pg/ml using silver enhancement.
- Assay performance was directly proportional to ricin concentration, indicated by color intensity.
Conclusions:
- A rapid and sensitive immunochromatographic assay for ricin detection was successfully developed.
- The assay offers a promising tool for quick and reliable ricin identification.
- Further optimization, including silver enhancement, can significantly boost assay sensitivity.
Abstract:
A rapid immunochromatographic assay was developed to detect ricin. The assay was based on the sandwich format using monoclonal antibodies (Mabs) of two distinct specificities. One anti-ricin B chain Mab (1G7) was immobilized to a defined detection zone on a porous nitrocellulose membrane, while the other anti-ricin A chain Mab (5E11) was conjugated to colloidal gold particles which served as a detection reagent. The ricin-containing sample was added to the membrane and allowed to react with Mab (5E11)-coated particles. The mixture was then passed along the porous membrane by capillary action past the Mab (1G7) in the detection zone, which will bind the particles that had ricin bound to their surface, giving a red color within this detection zone with an intensity proportional to ricin concentration. In the absence of ricin, no immunogold was bound to the solid-phase antibody. With this method, 50 ng/ml of ricin was detected in less than 10 min. The assay sensitivity can be increased by silver enhancement to 100 pg/ml.
