Methods and kits

A method for detecting the presence of minichromosome maintenance complex component 5 (Mcm5) in a subject, the method comprising:

• (a) preparing a sample from said subject wherein preparing the sample comprises exposing the sample to a lysis buffer, wherein the lysis buffer is capable of releasing Mcm5 from cells in the sample;
• (b) performing an assay to determine the concentration of Mcm5 by exposing the sample to a first monoclonal antibody and/or a second monoclonal antibody and measuring the amount of Mcm5 that binds to the first monoclonal antibody and/or the second monoclonal antibody;
• and
• (c) comparing the concentration of Mcm5 determined in step (b) to reference values;
• wherein the first monoclonal antibody and the second monoclonal antibody bind to Mcm5,
• wherein the first monoclonal antibody binds to a polypeptide comprising an amino acid sequence of SEQ ID NO:
• 1, and the second monoclonal antibody binds to a polypeptide comprising an amino acid sequence of SEQ ID NO:
• 2.

A kit comprising a lysis buffer which is capable of releasing Mcm5 from cells in a sample, a first monoclonal antibody and a second monoclonal antibody wherein the first monoclonal antibody is an antibody which:

• (i) binds to a polypeptide having an amino acid sequence of SEQ ID NO:
• 1;
• (ii) comprises at least one Complementary Determining Region (CDR) selected from the group consisting of:
• (a) 12A7 CDRH1 which has a sequence consisting of SEQ ID NO:
• 9 or a sequence that differs from SEQ ID NO:9 by a single amino acid substitution;
• (b) 12A7 CDRH2 which has a sequence consisting of SEQ ID NO:
• 11 or a sequence that differs from SEQ ID NO:11 by a single amino acid substitution;
• (c) 12A7 CDRH3 which has a sequence consisting of SEQ ID NO:
• 13 or a sequence that differs from SEQ ID NO:13 by a single amino acid substitution;
• (d) 12A7 CDRL1 which has a sequence consisting of SEQ ID NO:
• 3 or a sequence that differs from SEQ ID NO:3 by a single amino acid substitution;
• (e) 12A7 CDRL2 which has a sequence consisting of SEQ ID NO:
• 5 or a sequence that differs from SEQ ID NO:5 by a single amino acid substitution;
• and
• (f) 12A7 CDRL3 which has a sequence consisting of SEQ ID NO:
• 7 or a sequence that differs from SEQ ID NO:7 by a single amino acid substitution;
• (iii) comprises a heavy chain variable region having a sequence at least 85%, 90%, 95%, 98%, 99% or 100% identical to SEQ ID NO:
• 29;
• (iv) comprises a light chain variable region sequence having a sequence at least 85%, 90%, 95%, 98%, 99% or 100% identical to SEQ ID NO:
• 27;
• or
• (v) competes with the antibody of (i), (ii), (iii) or (iv);
• and the second monoclonal antibody is an antibody which:
• (i) binds to a polypeptide having an amino acid sequence of SEQ ID NO:
• 2;
• (ii) comprises at least one Complementary Determining Region (CDR) selected from the group consisting of:
• (a) 4B4 CDRH1 which has a sequence consisting of SEQ ID NO:
• 21 or a sequence that differs from SEQ ID NO:21 by a single amino acid substitution;
• (b) 4B4 CDRH2 which has a sequence consisting of SEQ ID NO:
• 23 or a sequence that differs from SEQ ID NO:23 by a single amino acid substitution;
• (c) 4B4 CDRH3 which has a sequence consisting of SEQ ID NO:
• 25 or a sequence that differs from SEQ ID NO:25 by a single amino acid substitution;
• (d) 4B4 CDRL1 which has a sequence consisting of SEQ ID NO:
• 15 or a sequence that differs from SEQ ID NO:15 by a single amino acid substitution;
• (e) 4B4 CDRL2 which has a sequence consisting of SEQ ID NO:
• 17 or a sequence that differs from SEQ ID NO:17 by a single amino acid substitution;
• and
• (f) 4B4 CDRL3 which has a sequence consisting of SEQ ID NO:
• 19 or a sequence that differs from SEQ ID NO:19 by a single amino acid substitution;
• (iii) comprises a heavy chain variable region having a sequence at least 85%, 90%, 95%, 98%, 99% or 100% identical to SEQ ID NO:
• 33;
• (iv) comprises a light chain variable region sequence having a sequence at least 85%, 90%, 95%, 98%, 99% or 100% identical to SEQ ID NO:
• 31;
• or
• (v) competes with the antibody of (i), (ii), (iii) or (iv).