Composition comprising anti-gliadin antibodies and method of use thereof to detect gliadin in a sample
The present invention provides anti-gliadin antibodies and antibody fragments, and polypeptides encoding the antibodies or fragment. Also disclosed are methods and kits for the use of such antibodies, fragments, or polypeptides in detection of gliadin. Further provided are heavy chain and light chain variable sequences and associated sequences of complementarity-determining regions (CDRs).
1 . A composition comprising an isolated antibody or antibody fragment comprising:
(1) a heavy chain variable region comprising a CDR1 of SEQ ID NO:3, a CDR2 of SEQ ID NO:4, and a CDR3 of SEQ ID NO:5 and a light chain variable region comprising a CDR1 of SEQ ID NO:8, a CDR2 of SEQ ID NO:9, and a CDR3 of SEQ ID NO:10; or
(2) a heavy chain variable region comprising a CDR1 of SEQ ID NO:13, a CDR2 of SEQ ID NO:14, and a CDR3 of SEQ ID NO: 15 and a light chain variable region comprising a CDR1 of SEQ ID NO: 18, a CDR2 of SEQ ID NO:19, and a CDR3 of SEQ ID NO:20;
wherein the antibody or antibody fragment binds to gliadin at an epitope defined by the amino acid sequence set forth in any one of SEQ ID NOs:21-32; and
wherein the antibody or antibody fragment is linked to a solid support.
2 . The composition of claim 1 , wherein the heavy chain variable region has an amino acid sequence of SEQ ID NO:2; and
wherein the light chain variable region has an amino acid sequence of SEQ ID NO:7.
3 . The composition of claim 1 , wherein the heavy chain variable region has an amino acid sequence of SEQ ID NO: 12; and
wherein the light chain variable region has an amino acid sequence of SEQ ID NO: 17.
4 . The composition of claim 1 , wherein the solid support is one of a microtiter plate, a microchip, or a bead.
5 . The composition of claim 1 , wherein the gliadin is wheat gliadin, barley hordein, or rye secalin.
6 . The composition of claim 1 , wherein the antibody or antibody fragment is linked to a detectable molecule.
7 . The composition of claim 1 , wherein the isolated antibody or antibody fragment is a first isolated antibody or antibody fragment and further comprising a second isolated antibody or antibody fragment;
wherein the first isolated antibody or antibody fragment comprises a heavy chain variable region comprising a CDR1 of SEQ ID NO:3, a CDR2 of SEQ ID NO:4, and a CDR3 of SEQ ID NO:5 and a light chain variable region comprising a CDR1 of SEQ ID NO:8, a CDR2 of SEQ ID NO:9, and a CDR3 of SEQ ID NO:10;
wherein the second isolated antibody or antibody fragment comprises a heavy chain variable region comprising a CDR1 of SEQ ID NO:13, a CDR2 of SEQ ID NO:14, and a CDR3 of SEQ ID NO: 15 and a light chain variable region comprising a CDR1 of SEQ ID NO: 18, a CDR2 of SEQ ID NO:19, and a CDR3 of SEQ ID NO:20;
wherein the first isolated antibody or antibody fragment and the second isolated antibody or antibody fragment bind to gliadin at an epitope defined by the amino acid sequence set forth in any one of SEQ ID NOs:21-32; and
wherein the first isolated antibody or antibody fragment and the second isolated antibody or antibody fragment are linked to the solid support.
8 . The composition of claim 7 , wherein the first isolated antibody or antibody fragment comprises the heavy chain variable region having an amino acid sequence of SEQ ID NO:2, and the light chain variable region having an amino acid sequence of SEQ ID NO:7.
9 . The composition of claim 7 , wherein the second isolated antibody or antibody fragment comprises the heavy chain variable region having an amino acid sequence of SEQ ID NO: 12, and the light chain variable region having an amino acid sequence of SEQ ID NO: 17.
10 . A kit for detecting gliadin comprising an antibody or antibody fragment of claim 1 .
11 . The kit of claim 10 , further comprising a solution for dissolving a sample.
12 . The kit of claim 11 , wherein the sample is a food or beverage sample.
13 . The kit of claim 10 , further comprising a positive control containing gliadin.
14 . The kit of claim 10 , further comprising a container.
15 . The kit of claim 10 , wherein the solid support is one of a microtiter plate, a microchip, or a bead.
16 . A method of detecting gliadin in a sample, the method comprising contacting the sample with the composition of claim 1 .
17 . The method of claim 16 , wherein the sample is a food or beverage sample.
18 . The method of claim 16 , further comprising dissolving the sample in a solution.
19 . The method of claim 18 , wherein the contacting the sample with the composition of claim 1 occurs after dissolving the sample in the solution.
20 . The method of claim 16 , wherein the gliadin includes wheat gliadin, barley hordein, or rye secalin.