Immunoassay for phenethylamines of the 2C and DO sub-families
Inventors
Benchikh, Elouard • Fitzgerald, Peter • McConnell, Ivan • Lowry, Philip
Assignees
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Publication Number
US-10775394-B2
Publication Date
2020-09-15
Expiration Date
Abstract
Immunoassay methods and their requisite components for the detection and determination of phenethylamines of the 2C and DO sub-families are described.
Core Innovation
The disclosed immunoassays provide first known immunoassays for selective detection of phenethylamine 2C and DO sub-families using novel sub-family-specific antibodies that do not cross-react across the two sub-families. The core problem addressed is the need for selective detection of phenethylamines within a 2C sub-family while avoiding cross-reactivity with related compounds from the opposite sub-family. The patent describes competitive immunoassay use for measuring the presence or amount of target phenethylamines.
The disclosed antibody system specifies polyclonal antibodies that specifically bind multiple listed 2C-subfamily phenethylamine compounds, including 2C-B, 2C-P, 2C-E, 2C-T2, 2C-C, 2C-T-7, 2C-D, and 2C-CN. The polyclonal antibodies are characterized by cross-reactivity constraints, including cross-reactivity of less than 1.0% for each of several listed related compounds compared to 100% cross-reactivity for 2C-B.
The disclosure further supports competitive immunoassay quantification using calibration curves and detecting agents, with signals measured from detecting agents to determine presence or amount. Optional solid substrates, including planar biochips with spatially addressable or multiplex screening, are described for immobilized antibodies.
Claims Coverage
The independent claim in the provided set defines a polyclonal antibody with binding specificity to multiple 2C-subfamily phenethylamines and a cross-reactivity threshold (<1.0%) for multiple other listed compounds, with cross-reactivity measured in the presence of a conjugate and the antibody derived from an immunogen of specified structure. The provided claim set indicates additional dependent limitations involving sensitivity, competitive immunoassay operation with calibration curves, sample matrix, and inclusion of a second polyclonal antibody on a substrate with its own binding/cross-reactivity constraints.
Polyclonal antibody specific binding to multiple 2C-subfamily phenethylamines
A polyclonal antibody which specifically binds to 1-(4-bromo-2,5-dimethoxyphenyl)ethan-2-amine (2C-B), 1-(2,5-dimethoxy-4-propylphenyl)ethan-2-amine (2C-P), 1-(2,5-dimethoxy-4-ethylphenyl)ethan-2-amine (2C-E), 1-(2,5-dimethoxy-4-ethylthiophenyl)ethan-2-amine (2C-T2), 1-(4-chloro-2,5-dimethoxyphenyl)ethan-2-amine (2C-C), 1-(2,5-dimethoxy-4-propylthiophenyl)ethan-2-amine (2C-T-7), 1-(2,5-dimethoxy4-methylphenyl)ethan-2-amine (2C-D), and 1-(4-cyano-2,5-dimethoxyphenyl)ethan-2-amine (2C-CN).
Cross-reactivity threshold measured with a conjugate
The polyclonal antibody shows a cross-reactivity of less than 1.0% for each of 1-(4-bromo-2,5-dimethoxyphenyl)propan-2-amine (DOB), (+)-Phenethylamine, Dimethylamylamine, when compared to 100% cross-reactivity for 2C-B, wherein said cross reactivity is measured in the presence of a conjugate of structure.
Antibody derived from a specified immunogen structure
The polyclonal antibody is derived from an immunogen of structure.
The independent claim coverage is directed to a polyclonal antibody that specifically binds multiple listed 2C-subfamily phenethylamines, while exhibiting less than 1.0% cross-reactivity for each of the listed comparator compounds compared to 100% cross-reactivity for 2C-B, with cross-reactivity measured in the presence of a conjugate and the antibody derived from an immunogen of specified structure. The provided claim set also includes dependent limitations involving calibration curves, sample types, and a substrate carrying a second polyclonal antibody.
Stated Advantages
Selective detection of phenethylamines within a 2C sub-family using sub-family-specific antibodies that do not cross-react across the two sub-families.
Low cross-reactivity (<1.0%) for listed compounds compared to 100% cross-reactivity for 2C-B (as measured with a conjugate).
Competitive immunoassay format supports determination of presence or amount using a calibration curve.
Quantified sensitivity is provided for 2C-B (IC50 value reported in dependent claim listing).
Supports detection using specified biological sample types (blood, serum, plasma, urine, or saliva) as stated in dependent claim listing.
Documented Applications
Immunoassay-based detection of phenethylamines of a 2C sub-family in biological samples including blood, serum, plasma, urine, or saliva.
Use of an immunoassay competitive assay format where phenethylamines compete for binding and signals are measured to infer presence or amount using a calibration curve.
Substrate-based detection using a substrate including immobilized antibodies, with an example of a second polyclonal antibody on the same substrate for binding DOB, DOM, and DON with defined cross-reactivity constraints.
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