Abstract A determination of the presence and amount of a specific organic substance (ligand) that will form a complex with a macromolecule (antibody) may be made by means of a reagent obtained by binding to the ligand to be assayed a chemiluminescent substance. The addition of a limited amount of a substance (antibody) having receptors for the ligand together with the chemiluminescent labeled ligand to the fluid to be assayed results in a competitive reaction between the ligand present in the fluid and the chemiluminescent labeled ligand for the limited number of receptor sites. Under equilibrium conditions, the amount of chemiluminescent labeled ligand bound to the antibody is related to the amount of unlabeled ligand in the solution being assayed, and is determined by isolating the antibody and measuring its chemiluminescence or by isolating and measuring the amount of free labeled-ligand remaining.
An insulin-chemiluminescent substance conjugate having luminescent activity measurable by photosensitive means.
Claim 2
Dependent Claim
11 words
An insulin-chemiluminescent substance conjugate having luminescent activity measurable by photosensitive means.
Claim 3
Dependent Claim
87 words
The insulin-chemiluminescent conjugate of claim 1 wherein said chemiluminescent substance is selected from the group consisting of 2,3-dihydropthalizine-1,4-diones of the following general formula: ##STR4## where R is: NH 2 ; NCH 3 ; NHC 2 H 5 ; NHCH 2 C 6 H 5 ; NHCOCH 2 Cl; NHCOCH 2 NH 2 ; NHCOCH 2 NHNH 2 ; N(CH 3 ) 2 ; N(C 2 H 5 ) 2 ; N(CH 2 C 6 H 5 ) 2 , tetrabis (dimethylamine) ethylene, luciferin, lucigenin or oxalyl chloride.
Claim 4
Dependent Claim
17 words
The ligand-chemiluminescent conjugate of claim 1 wherein said ligand is insulin and said chemiluminescent substance is luminol.
Claim 5
Dependent Claim
14 words
An insulin-luminol conjugate prepared by reacting luminol with insulin in the presence of glutaraldehyde.
Claim 6
Dependent Claim
14 words
An insulin-luminol conjugate prepared by reacting luminol with insulin in the presence of glutaraldehyde.
Claim 7
Dependent Claim
13 words
The insulin-luminol conjugate of claim 4 having a label ratio of about 1:10.
Claim 8
Independent Claim
21 words
The method of activating an insulin-luminol conjugate which comprises adding thereto a solution containing an alkali metal ferricyanide and hydrogen peroxide.
Claim 9
Independent Claim
21 words
The method of activating an insulin-luminol conjugate which comprises adding thereto a solution containing an alkali metal ferricyanide and hydrogen peroxide.
Claim 10
Independent Claim
15 words
The method of activating an insulin-luminol conjugate which comprises adding thereto an alkali metal hypochlorite.
Claim 11
Independent Claim
15 words
The method of activating an insulin-luminol conjugate which comprises adding thereto an alkali metal hypochlorite.