Methods of preparing
Disclosed are methods related to the generation and use of cellular and humoral responses for the prevention and treatment of P. gingivalis related conditions and diseases, such as methods of preparing P. gingivalis antibodies, comprising immunizing a non-human animal with a chimeric or fusion protein, wherein the protein comprises a first peptide joined directly or through a linker to a second peptide or polypeptide, wherein (A) the first peptide comprises a region of a P. gingivalis trypsin-like enzyme and (B) the second peptide or polypeptide comprises an adhesin domain of P. gingivalis.
1. A method of preparing an antibody, comprising immunizing a non-human animal with a chimeric or fusion protein, wherein the protein comprises a first peptide joined directly or through a linker to a second peptide or polypeptide, wherein:
(A) said first peptide comprises a region of a P. gingivalis trypsin-like enzyme selected from the group consisting of the amino acid sequence that is the same as, or at least 90% homologous to, the sequence shown in any one of SEQ ID NOs: 1-34; and
(B) said second peptide or polypeptide comprises a P. gingivalis adhesin domain sequence selected from the group consisting of:
(i) the amino acid sequence that is the same as, or at least 90% homologous to, the P. gingivalis Lys-X-proteinase adhesin domain sequence shown in any one of SEQ ID NOs: 35-37 and 40-43 and;
(ii) the amino acid sequence that is the same as, or at least 90% homologous to, the P. gingivalis Arg-X-proteinase adhesin domain sequence shown in any one of SEQ ID NOs: 38-39 and 44-46; and
(iii) the amino acid sequence that is the same as, or at least 90% homologous to, the of P. gingivalis HagA adhesin domain sequence shown in any one of SEQ ID NOs: 80-82; and
(iv) the amino acid sequence that is the same as, or at least 90% homologous to, the sequence shown in any one of SEQ ID NOs: 69-79 and 83-85.
2. The method of claim 1 wherein said first peptide comprises a sequence selected from the group consisting of SEQ ID NOs: 3-34 and sequences at least 90% homologous thereto.
3. The method of claim 1 wherein said second peptide or polypeptide comprises a sequence selected from the group consisting of SEQ ID NOs: 35-39 and sequences at least 90% homologous thereto.
4. The method of claim 1 , wherein:
(a) said first peptide comprises a sequence that is the same as or at least 90% homologous to a sequence selected from the group consisting of SEQ ID NOs: 27-30 and said second peptide or polypeptide comprises a sequence that is the same as or at least 90% homologous to a sequence selected from the group consisting of SEQ ID NOs: 36-37; or
(b) said first peptide comprises a sequence that is the same as or at least 90% homologous to a sequence selected from the group consisting of SEQ ID NOs: 31-34 and said second peptide or polypeptide comprises a sequence that is the same as or at least 90% homologous to SEQ ID NO:39.
5. The method of claim 1 wherein:
the C-terminal residue of said first peptide is covalently linked directly or through a linker that is either (i) up to 15 amino acids in length, or (ii) less than 5 amino acids in length, to either (a) the N-terminal residue or (b) the C-terminal residue of said second peptide or polypeptide, or
the N-terminal residue of said first peptide is covalently linked directly or through a linker that is either (i) up to 15 amino acids in length, or (ii) less than 5 amino acids in length, to either (a) the N-terminal residue or (b) the C-terminal residue of said second peptide or polypeptide.
6. The method of claim 1 wherein the first peptide comprises a sequence selected from the group consisting of SEQ ID Nos: 27-30, and sequences at least 90% homologous thereto, and said second polypeptide comprises a sequence selected from SEQ ID NO: 36-37, and sequences at least 90% homologous thereto.
7. The method of claim 1 wherein said first peptide comprises a sequence that is the same as or is at least 90% identical to the sequence of SEQ ID NO: 28 and said second peptide or polypeptide comprises a sequences that is the same or is at least 90% identical to the sequence of SEQ ID NO: 37.
8. The method of claim 1 wherein said first peptide comprises a sequence that is the same as or is at least 90% identical to the sequence of SEQ ID NO: 27 and said second peptide or polypeptide comprises a sequences that is the same or is at least 90% identical to the sequence of SEQ ID NO: 36.