Patent ID: 6730482
Filing Date: 2004-05-04
Classification: C12N,C12Q

Abstract:
A method for generating nucleic acid ligands to components of a biological tissue or fluid present under a first predetermined condition and not present under a second predetermined condition, comprising:a) providing a peptide, said peptide comprising a linear amino acid sequence identical to at least a portion of a protein suspected of being a component of said biological tissue or fluid obtained under said first predetermined condition; b) providing a candidate mixture of nucleic acids; c) contacting the candidate mixture of nucleic acids with said peptide, wherein nucleic acids having an increased affinity to said peptide relative to the candidate mixture may be partitioned from the remainder of the candidate mixture; d) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture; e) amplifying the increased affinity nucleic acids to yield a candidate mixture of nucleic acids enriched for nucleic acids with relatively higher affinity and specificity for said peptide, whereby nucleic acids having a relatively higher affinity for said biological tissue or fluid obtained under said first predetermined condition are generated; f) contacting said enriched candidate mixture with said biological tissue or fluid obtained under said first predetermined condition, wherein nucleic acids having an increased affinity to components of said biological tissue or fluid present under said first predetermined condition relative to the candidate mixture may be partitioned from the remainder of the candidate mixture; g) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture; h) contacting the increased affinity nucleic acids with said biological tissue or fluid obtained under said second predetermined condition, wherein nucleic acids with affinity to components of said biological tissue or fluid present under a second predetermined condition are removed from said increased affinity nucleic acids, whereby nucleic nucleic acids having a relatively higher affinity for said biological tissue or fluid obtained under said first predetermined condition and depleted for nucleic acids with relatively higher affinity and specificity for components of said biological tissue or fluid present under said second predetermined condition are generated; and i) amplifying the nucleic acids retained in step h); whereby nucleic acid ligands to components of said biological tissue or fluid present under said first predetermined condition and not present under said second predetermined condition may be identified.