Assay for detecting, measuring and monitoring the activities and concentrations of proteins and methods of use thereof

A method of detecting or confirming whether a subject was exposed to an agent which affects the concentration or activity of acetylcholinesterase, butyrylcholinesterase, or both comprising

• obtaining a test sample from the subject;
• measuring the reaction rates between acetylcholinesterase and a plurality of substrates;
• measuring the reaction rates between butyrylcholinesterase and the plurality of substrates;
• and
• calculating the activity or the concentration of acetylcholinesterase, butyrylcholinesterase, or both with sensitivity coefficients of each substrate for acetylcholinesterase and butyrylcholinesterase.

A method of determining the identity of an agent which affects the concentration or activity of acetylcholinesterase, butyrylcholinesterase, or both to which a subject was exposed comprising

• obtaining a test sample from the subject;
• measuring the reaction rates between acetylcholinesterase and a plurality of substrates;
• measuring the reaction rates between butyrylcholinesterase and the plurality of substrates;
• and
• calculating the activity or the concentration of acetylcholinesterase, butyrylcholinesterase, or both with sensitivity coefficients of each substrate for acetylcholinesterase and butyrylcholinesterase;
• and
• comparing the activities or the concentrations with a database of activity and concentration acetylcholinesterase and butyrylcholinesterase profiles for agents which affect the concentration or activity of acetylcholinesterase, butyrylcholinesterase, or both.

A method of determining the efficacy or monitoring the progress of a treatment regime, wherein a subject is administered a compound which affects the concentration or activity of acetylcholinesterase, butyrylcholinesterase, or both comprising

• obtaining a test sample from the subject;
• measuring the reaction rates between acetylcholinesterase and a plurality of substrates;
• measuring the reaction rates between butyrylcholinesterase and the plurality of substrates;
• calculating the activity or the concentration of acetylcholinesterase, butyrylcholinesterase, or both with sensitivity coefficients of each substrate for acetylcholinesterase and butyrylcholinesterase;
• and
• monitoring the activities or the concentrations of acetylcholinesterase, butyrylcholinesterase, or both as a function of time of the treatment regime.

A method of determining whether a subject suffers from a drug sensitivity or a disease which affects the activities or the concentrations of acetylcholinesterase, butyrylcholinesterase, or both comprising

• obtaining a test sample from the subject;
• measuring the reaction rates between acetylcholinesterase and a plurality of substrates;
• measuring the reaction rates between butyrylcholinesterase and the plurality of substrates;
• calculating the activity or the concentration of acetylcholinesterase, butyrylcholinesterase, or both with sensitivity coefficients of each substrate for acetylcholinesterase and butyrylcholinesterase;
• and
• comparing the activities or the concentrations with a database of activity and concentration acetylcholinesterase and butyrylcholinesterase profiles which are typical of individuals suffering from given drug sensitivities and individuals suffering from given diseases which affect the activities or the concentrations of acetylcholinesterase, butyrylcholinesterase, or both.

A method of measuring the concentration of red blood cells in a subject comprising

• obtaining a test sample from the subject;
• measuring the reaction rates between acetylcholinesterase and a plurality of substrates;
• measuring the reaction rates between butyrylcholinesterase and the plurality of substrates;
• calculating the activity or the concentration of acetylcholinesterase, butyrylcholinesterase, or both with sensitivity coefficients of each substrate for acetylcholinesterase and butyrylcholinesterase;
• determining a relationship between standard concentrations of red blood cells and the activities or the concentrations of acetylcholinesterase, butyrylcholinesterase, or both;
• and
• using the relationship to calculate the concentration of red blood cells of the sample.

A method of screening for a candidate compound which affects the concentration or activity of acetylcholinesterase, butyrylcholinesterase, or both comprising

• obtaining a test sample;
• measuring the reaction rates between acetylcholinesterase and a plurality of substrates;
• measuring the reaction rates between butyrylcholinesterase and the plurality of substrates;
• calculating the activity or the concentration of acetylcholinesterase, butyrylcholinesterase, or both with sensitivity coefficients of each substrate for acetylcholinesterase and butyrylcholinesterase;
• and
• determining whether the concentration or activity of acetylcholinesterase, butyrylcholinesterase, or both changes.

A database of sensitivity coefficients for calculating the activities or the concentrations of acetylcholinesterase, butyrylcholinesterase, or both made by a method comprising

• obtaining a plurality of inhibited dilutions of a sensitivity coefficient sample, wherein the plurality of inhibited dilutions comprise a plurality of concentrations of either acetylcholinesterase or butyrylcholinesterase which is partially to completely inhibited;
• exposing each inhibited dilution of the plurality of inhibited dilutions to each substrate in a plurality of substrates for acetylcholinesterase and butyrylcholinesterase;
• measuring the reaction rates between acetylcholinesterase and each substrate;
• measuring the reaction rates between butyrylcholinesterase and each substrate;
• calculating the relationship between the reaction rates of acetylcholinesterase and each concentration of the sensitivity coefficient sample at infinite inhibitor concentration;
• calculating the relationships between the reaction rates of butyrylcholinesterase and each concentration of the sensitivity coefficient sample at infinite inhibitor concentration;
• and
• extracting each sensitivity coefficient of each substrate for acetylcholinesterase and butyrylcholinesterase from the calculated relationships.