Antibodies to quetiapine haptens and use thereof
Disclosed is an antibody which binds to quetiapine, which can be used to detect quetiapine in a sample such as in a competitive immunoassay method. The antibody can be used in a lateral flow assay device for point-of-care detection of quetiapine, including multiplex detection of aripiprazole, olanzapine, quetiapine, and risperidone in a single lateral flow assay device.
1. A method of producing an antibody or a binding fragment thereof which specifically binds to quetiapine, the method comprising:
(i) selecting a host for antibody production; and
(ii) inoculating the host with a conjugate of a compound of Formula I and an immunogenic carrier; and
(iii) isolating an antibody or binding fragment thereof which specifically binds to quetiapine produced by the host,
Formula I:
wherein:
R 1 is H,
CH 2 NH 2 , or CH 2 NHC(O)(CH 2 ) m CO 2 H;
R 2 is H,
CH 2 NH 2 , or CH 2 NHC(O)(CH 2 ) m CO 2 H;
R 3 is H; provided that either R 1 or R 2 must be H, and further provided that both R 1 and R 2 may not be H simultaneously;
m is 1, 2, 3, 4, or 5; and
n is 1, 2, 3, 4, or 5, and wherein the immunogenic carrier is conjugated to the compound of Formula I at R 1 when R 1 is not H, or at R 2 when R 2 is not H.
2. A method of producing a hybridoma cell line capable of producing a monoclonal antibody which binds to quetiapine, the method comprising:
(i) selecting a host for antibody production;
(ii) inoculating the host with a conjugate of a compound of Formula I and an immunogenic carrier;
(iii) fusing a cell line capable of producing an antibody that binds to quetiapine from said inoculated host with an immortalized cell to create a fused cell capable of producing a monoclonal antibody which binds to quetiapine; and
(iv) cloning the fused cell so as to obtain a hybridoma cell line, Formula I:
wherein:
CH 2 NH 2 , or CH 2 NHC(O)(CH 2 ) m CO 2 H;
R 2 is H,
CH 2 NH 2 , or CH 2 NHC(O)(CH 2 ) m CO 2 H;
R 3 is H; provided that either R 1 or R 2 must be H, and further provided that both R 1 and R 2 may not be H simultaneously;
m is 1, 2, 3, 4, or 5; and
n is 1, 2, 3, 4, or 5, and wherein the immunogenic carrier is conjugated to the compound of Formula I at R 1 when R 1 is not H, or at R 2 when R 2 is not H.
3. The method of claim 1 , wherein:
R 1 is H,
or CH 2 NH 2 ;
R 2 is H,
or CH 2 NH 2 ,
R 3 is H; provided that either R 1 or R 2 must be H, and further provided that both R 1 and R 2 may not be H simultaneously;
m is 1, 2, 3, 4, or 5; and
n is 1, 2, 3, 4, or 5.
4. The method of claim 1 , wherein
R 1 is H, or CH 2 NHC(O)(CH 2 ) m CO 2 H;
R 2 is H, or CH 2 NHC(O)(CH 2 ) m CO 2 H;
R 3 is H; provided that either R 1 or R 2 must be H, and further provided that both R 1 and R 2 may not be H simultaneously;
m is 1, 2, 3, 4, or 5; and
n is 1, 2, 3, 4, or 5.
5. The method of claim 1 , wherein:
R 1 is H,
or CH 2 NHC(O)(CH 2 ) m CO 2 H;
R 2 is H,
or CH 2 NHC(O)(CH 2 ) m CO 2 H;
R 3 is H; provided that either R 1 or R 2 must be H, and further provided that R 1 and R 2 may not be H simultaneously;
m is 1, 2, 3, 4, or 5; and
n is 1, 2, 3, 4, or 5.
6. The method of claim 1 , wherein the compound of Formula I is selected from the group consisting of:
wherein:
m is 1, 2, 3, 4, or 5; and
n is 1, 2, 3, 4, or 5.
7. The method of claim 2 , wherein
R 1 is H,
or CH 2 NH 2 ;
R 2 is H,
or CH 2 NH 2 ,
R 3 is H; provided that either R 1 or R 2 must be H, and further provided that both R 1 and R 2 may not be H simultaneously;
m is 1, 2, 3, 4, or 5; and
n is 1, 2, 3, 4, or 5.
8. The method of claim 2 , wherein
R 1 is H, or CH 2 NHC(O)(CH 2 ) m CO 2 H;
R 2 is H, or CH 2 NHC(O)(CH 2 ) m CO 2 H;
R 3 is H; provided that either R 1 or R 2 must be H, and further provided that both R 1 and R 2 may not be H simultaneously;
m is 1, 2, 3, 4, or 5; and
n is 1, 2, 3, 4, or 5.
9. The method of claim 2 , wherein:
R 1 is H,
or CH 2 NHC(O)(CH 2 ) m CO 2 H;
R 2 is H,
or CH 2 NHC(O)(CH 2 ) m CO 2 H;
R 3 is H; provided that either R 1 or R 2 must be H, and further provided that R 1 and R 2 may not be H simultaneously;
m is 1, 2, 3, 4, or 5; and
n is 1, 2, 3, 4, or 5.
10. The method of claim 2 , wherein the compound of Formula I is selected from the group consisting of:
wherein:
m is 1, 2, 3, 4, or 5; and
n is 1, 2, 3, 4, or 5.
11. The method of claim 1 , wherein the immunogenic carrier is a protein.
12. The method of claim 11 , wherein the protein is selected from keyhole limpet hemocyanin, bovine thyroglobulin, and ovalbumin.
13. The method of claim 2 , wherein the immunogenic carrier is a protein.
14. The method of claim 13 , wherein the protein is selected from keyhole limpet hemocyanin, bovine thyroglobulin, and ovalbumin.