Anti-gliadin antibodies, encoding nucleic acids and methods of detecting gliadin
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. 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.
2. The isolated antibody or antibody fragment 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 isolated antibody or antibody fragment 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 isolated antibody or antibody fragment of claim 1 , wherein the gliadin is wheat gliadin, barley hordein, or rye secalin.
5. The isolated antibody or antibody fragment of claim 1 , wherein the antibody is a single-chain Fv (scFV).
6. The isolated antibody or antibody fragment of claim 1 , wherein the antibody is an IgG.
7. The isolated antibody or antibody fragment of claim 1 , wherein the antibody is linked to a detectable agent.
8. A kit for detecting gliadin comprising an antibody or antibody fragment of claim 1 .
9. The kit of claim 8 , further comprising a solution for dissolving a test sample.
10. The kit of claim 8 , wherein the antibody or antibody fragment is linked to a detectable agent.
11. The kit of claim 8 , further comprising a positive control containing gliadin.
12. An isolated nucleic acid comprising a polynucleotide sequence encoding an antibody 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; an antibody 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; an antibody 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; or an antibody light chain variable region comprising a CDR1 of SEQ NO:18, a CDR2 of SEQ ID NO:19, and a CDR3 of SEQ ID NO:20.
13. The nucleic acid of claim 12 , wherein the polynucleotide sequence encodes (1) an antibody heavy chain variable region having the amino acid sequence of SEQ ID NO:2 or 12; or (2) an antibody light chain variable region having the amino acid sequence of SEQ ID NO:7 or 17.
14. The nucleic acid of claim 12 , wherein the polynucleotide sequence is SEQ ID NO:1, 6, 11, or 16.
15. The nucleic acid of claim 14 , further comprising a promoter operably linked to the polynucleotide sequence.
16. A method of detecting gliadin in a test sample comprising contacting the sample with the antibody or antibody fragment of claim 1 .
17. The method of claim 16 , wherein the sample is a food or beverage sample.