p53 PEPTIDES AS MARKERS IN THE DIAGNOSIS AND PROGNOSIS OF ALZHEIMER'S DISEASE
AssigneeDiadem S.r.l.
InventorsDaniela Letizia Uberti, Maurizio Memo
Life of this patent
Estimating the term…
Abstract
Disclosed are p53 peptides and their use as biomarkers in the diagnosis and/or prognosis of Alzheimer's disease (AD) in a biological sample. The invention also provides for a diagnostic method based on a highly accurate mass spectrometry analysis for the diagnosis of Alzheimer's disease at the pre-clinical and prodromal stages of the disease and for the prognosis of cognitive decline in a subject, by quantitating the levels of said p53 peptides specifically in human plasma of patients.
Claims51
A method for diagnosis of a subject as being cognitive normal (CN) or having mild cognitive impairment (MCI) or Alzheimer's disease (AD) or the prognosis of cognitive decline leading to dementia, the method comprising:
- subjecting isolated p53 protein to protease digestion to generate one or more peptides comprising one or both of peptides P1 and P2, said peptide P1 having (SEQ ID NO. 1 TEEENLR) and said peptide P2 having (SEQ ID NO. 2 TEEENLRK[GG]K), wherein said one or peptides are proteolytic products of said protease digestion,
- wherein said p53 protein is isolated from an immunoprecipitate obtained from a biological sample from said subject, said immunoprecipitate comprising an antibody specific for p53 protein and p53 protein present in said sample,
- wherein said antibody specific for p53 protein comprises a heavy chain variable region comprising CDR1 (SEQ ID NO:10), CDR2 (SEQ ID NO:
- 11) and CDR3 (SEQ ID NO:
- 12) and a light chain variable region comprising CDR1 (SEQ ID NO:13), CDR2 (SEQ ID NO:
- 14) and CDR3 (SEQ ID NO:
- 15);
- and
- comparing an amount of each of said one or more proteolytic peptides in a reaction mixture to an amount of each of one or more respective control peptides, wherein a higher amount of said one or more proteolytic peptides relative to said one or more respective control peptides indicates Alzheimer's disease or cognitive decline leading to dementia in said subject.
and CDR3 (SEQ ID NO:
- 15).
A method of providing a quantified reaction mixture comprising one or both of peptides P1 and P2, said peptide P1 having (SEQ ID NO. 1 TEEENLR) and said peptide P2 having (SEQ ID NO. 2 TEEENLRK[GG]K), the method comprising the steps of:
- subjecting a human p53 isoform that is conformationally altered with respect to the wild-type human p53 protein to protease digestion to generate said P1 peptide and/or P2 peptide,
- wherein said one or peptides are proteolytic products of said protease digestion,
- wherein said p53 isoform is isolated from an immunoprecipitate obtained from a biological sample from said subject, said immunoprecipitate comprising an antibody specific for p53 protein and p53 protein present in said sample,
- wherein said antibody specific for p53 protein comprises a heavy chain variable region comprising CDR1 (SEQ ID NO:10), CDR2 (SEQ ID NO:
- 11) and CDR3 (SEQ ID NO:
- 12) and a light chain variable region comprising CDR1 (SEQ ID NO:13), CDR2 (SEQ ID NO:
- 14) and CDR3 (SEQ ID NO:
- 15);
- isolating said proteolytic products to generate said reaction mixture comprising said P1 or P2 peptide and quantifying the amount of said P1 and P2 peptide in said reaction mixture, thereby providing said quantified reaction mixture.
A composition comprising a mixture of one or both of peptides P1 and P2, said peptide P1 having (SEQ ID NO. 1 TEEENLR) and said peptide P2 having (SEQ ID NO. 2 TEEENLRK[GG]K),
- wherein said one or peptides are proteolytic products of a protease digestion of an isolated human p53 protein,
- wherein said p53 protein is isolated from an immunoprecipitate obtained from a biological sample from a human subject, said immunoprecipitate comprising an antibody specific for p53 protein and p53 protein present in said sample,
- wherein said antibody specific for p53 protein comprises a heavy chain variable region comprising CDR1 (SEQ ID NO:10), CDR2 (SEQ ID NO:
- 11) and CDR3 (SEQ ID NO:
- 12) and a light chain variable region comprising CDR1 (SEQ ID NO:13), CDR2 (SEQ ID NO:
Dependencies are inferred from the claim text.