Antibody characterization and immunoassays for palytoxin using an SPR biosensor |
| |
Authors: | Yakes Betsy Jean DeGrasse Stacey L Poli Mark Deeds Jonathan R |
| |
Affiliation: | (1) Center for Food Safety and Applied Nutrition, US Food and Drug Administration, 5100 Paint Branch Parkway, College Park, MD 20740, USA;(2) Integrated Toxicology Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD 21702, USA |
| |
Abstract: | Palytoxin (PLTX), a polyether marine toxin originally isolated from the zoanthid Palythoa toxica, is one of the most toxic non-protein substances known. Fatal poisonings have been linked to ingestion of PLTX-contaminated
seafood, and effects in humans have been associated with dermal and inhalational exposure to PLTX containing organisms and
waters. Additionally, PLTX co-occurrence with other well-characterized seafood toxins (e.g., ciguatoxins, saxitoxins, tetrodotoxin)
has hindered direct associations of PLTX to seafood-borne illnesses. There are currently no validated methods for the quantitative
detection of PLTX(s). As such, a well-characterized, robust, specific analytical technique is needed for the detection of
PLTX(s) in source organisms, surrounding waters, and clinical samples. Surface plasmon resonance (SPR) biosensors are ideally
suited for antibody characterization and quantitative immunoassay detection. Herein, we describe a newly developed SPR assay
for PLTX. An anti-mouse substrate was used to characterize the kinetic values for a previously developed monoclonal anti-PLTX.
The characterized antibody was then incorporated into a sensitive, rapid, and selective PLTX assay. Buffer type, flow rate,
analyte-binding time, and regeneration conditions were optimized for the antibody–PLTX system. Cross-reactivity to potentially
co-occurring seafood toxins was also evaluated. We show that this optimized assay is capable of measuring low- to sub-ng/mL
PLTX levels in buffer and two seafood matrices (grouper and clam). Preliminary results indicate that this SPR biosensor assay
allows for (1) rapid characterization of antibodies and (2) rapid, sensitive PLTX concentration determination in seafood matrices.
Method development information contained herein may be broadly applied to future PLTX detection and/or antibody characterization
efforts. |
| |
Keywords: | |
本文献已被 PubMed SpringerLink 等数据库收录! |
|