Peptide mimotopes of claudin 18.2 and uses thereof
The present invention provides molecules that mimic antigenic determinants of the integral transmembrane protein claudin 18.2 (CLDN18.2). These molecules compete with CLDN18.2 for binding to a CLDN18.2 binding domain, e.g. a CLDN18.2 binding domain of an antibody, and are capable of detecting antibodies against CLDN18.2. The mimotopes of the invention may be used to generate or inhibit immune responses in animals and preferably humans. Furthermore, they can be used for purposes of detecting agents comprising a CLDN18.2 binding domain in biological samples as well as for purifying agents comprising a CLDN18.2 binding domain.
1. A peptide mimotope of claudin 18.2 (CLDN18.2) comprising the amino acid sequence
Cys Xaa1 Xaa2 Xaa3 Tyr Pro Gly Xaa4 Cys (SEQ ID NO: 19),
wherein
Xaa1 is an amino acid selected from the group consisting of Lys, Met, His, and Tyr,
Xaa2 is an amino acid selected from the group consisting of Leu, Lys, Tyr, Arg, and Phe,
Xaa3 is an amino acid selected from the group consisting of Gly, Asn, Arg, Ser, Trp, and Lys, and
Xaa4 is an amino acid selected from the group consisting of Trp, Tyr, Glu, Arg, Val, Lys, Ile, Met, and Phe,
wherein the thiol group of said cysteine residues form an intramolecular disulfide bridge.
2. The peptide mimotope of claim 1 , which comprises the amino acid sequence selected from the group consisting of:
Cys His Leu Gly Tyr Pro Gly Arg Cys (SEQ ID NO: 37),
Cys His Tyr Gly Tyr Pro Gly Arg Cys (SEQ ID NO: 38),
Cys His Leu Gly Tyr Pro Gly Trp Cys (SEQ ID NO: 39),
Cys His Tyr Ser Tyr Pro Gly Val Cys (SEQ ID NO: 40),
Cys His Tyr Gly Tyr Pro Gly Val Cys (SEQ ID NO: 41),
Cys His Tyr Ser Tyr Pro Gly Trp Cys (SEQ ID NO: 42),
Cys His Leu Arg Tyr Pro Gly Glu Cys (SEQ ID NO: 43),
Cys His Tyr Arg Tyr Pro Gly Glu Cys (SEQ ID NO: 44),
Cys His Leu Asn Tyr Pro Gly Tyr Cys (SEQ ID NO: 45),
Cys His Leu Gly Tyr Pro Gly Tyr Cys (SEQ ID NO: 46),
Cys His Leu Asn Tyr Pro Gly Trp Cys (SEQ ID NO: 47),
Cys Tyr Lys Gly Tyr Pro Gly Tyr Cys (SEQ ID NO: 48), and
Cys His Tyr Gly Tyr Pro Gly Trp Cys (SEQ ID NO: 49).
3. The peptide mimotope of claim 1 , wherein:
Xaa1 is an amino acid selected from the group consisting of His and Tyr;
Xaa2 is an amino acid selected from the group consisting of Leu, Lys, Tyr, and Phe;
Xaa3 is an amino acid selected from the group consisting of Gly, Asn, Arg, Ser, Trp, and Lys; and
Xaa4 is an amino acid selected from the group consisting of Trp, Tyr, Glu, Arg, Val, Lys, Ile, Met, and Phe.
4. The peptide mimotope of claim 1 , wherein:
Xaa1 is an amino acid selected from the group consisting of His and Tyr;
Xaa2 is an amino acid selected from the group consisting of Leu, Lys, and Tyr;
Xaa3 is an amino acid selected from the group consisting of Gly, Asn, Arg, and Ser; and
Xaa4 is an amino acid selected from the group consisting of Trp, Tyr, Glu, Arg, and Val.
5. The peptide mimotope of claim 1 , wherein:
Xaa1 is His;
Xaa2 is Tyr;
Xaa3 is Gly; and
Xaa4 is Val.
6. The peptide mimotope of claim 1 , wherein the peptide mimotope is characterized as a competitor of CLDN18.2 for binding to a CLDN18.2 binding domain.
7. The peptide mimotope of claim 1 , wherein the peptide mimotope is part of a fusion polypeptide.
8. The peptide mimotope of claim 1 , wherein the peptide mimotope is conjugated to at least one fusion partner.
9. The peptide mimotope of claim 8 , wherein the fusion partner comprises a heterologous amino acid sequence.
10. The peptide mimotope of claim 8 , wherein the fusion partner comprises a reporter for an immunological assay or a label.
11. A recombinant nucleic acid which encodes a peptide mimotope of claim 1 .
12. A host cell comprising the recombinant nucleic acid of claim 11 .
13. A peptide mimotope of claudin 18.2 (CLDN18.2) comprising the amino acid sequence
Ala Cys Xaa1 Xaa2 Xaa3 Tyr Pro Gly Xaa4 Cys Gly (SEQ ID NO: 21),
wherein
Xaa1 is an amino acid selected from the group consisting of Lys, Met, His, and Tyr;
Xaa2 is an amino acid selected from the group consisting of Leu, Lys, Tyr, Arg, and Phe;
Xaa3 is an amino acid selected from the group consisting of Gly, Asn, Arg, Ser, Trp, and Lys; and
Xaa4 is an amino acid selected from the group consisting of Trp, Tyr, Glu, Arg, Val, Lys, Ile, Met, and Phe,
wherein the thiol group of said cysteine residues form an intramolecular disulfide bridge.
14. The peptide mimotope of claim 13 , wherein:
Xaa1 is an amino acid selected from the group consisting of His and Tyr;
Xaa2 is an amino acid selected from the group consisting of Leu, Lys, and Tyr;
Xaa3 is an amino acid selected from the group consisting of Gly, Asn, Arg, and Ser; and
Xaa4 is an amino acid selected from the group consisting of Trp, Tyr, Glu, Arg, and Val.
15. The peptide mimotope of claim 13 , which comprises the amino acid sequence selected from the group consisting of:
Ala Cys His Leu Gly Tyr Pro Gly Arg Cys Gly (SEQ ID NO: 51),
Ala Cys His Tyr Gly Tyr Pro Gly Arg Cys Gly (SEQ ID NO: 52),
Ala Cys His Leu Gly Tyr Pro Gly Trp Cys Gly (SEQ ID NO: 53),
Ala Cys His Tyr Ser Tyr Pro Gly Val Cys Gly (SEQ ID NO: 54),
Ala Cys His Tyr Gly Tyr Pro Gly Val Cys Gly (SEQ ID NO: 55),
Ala Cys His Tyr Ser Tyr Pro Gly Trp Cys Gly (SEQ ID NO: 56),
Ala Cys His Leu Arg Tyr Pro Gly Glu Cys Gly (SEQ ID NO: 57),
Ala Cys His Tyr Arg Tyr Pro Gly Glu Cys Gly (SEQ ID NO: 58),
Ala Cys His Leu Asn Tyr Pro Gly Tyr Cys Gly (SEQ ID NO: 59),
Ala Cys His Leu Gly Tyr Pro Gly Tyr Cys Gly (SEQ ID NO: 60),
Ala Cys His Leu Asn Tyr Pro Gly Trp Cys Gly (SEQ ID NO: 61),
Ala Cys Tyr Lys Gly Tyr Pro Gly Tyr Cys Gly (SEQ ID NO: 62), and
Ala Cys His Tyr Gly Tyr Pro Gly Trp Cys Gly (SEQ ID NO: 63).