×

Systematic evolution of ligands by exponential enrichment: chimeric selex

  • US 5,637,459 A
  • Filed: 08/02/1994
  • Issued: 06/10/1997
  • Est. Priority Date: 06/11/1990
  • Status: Expired due to Term
First Claim
Patent Images

1. A method of identifying chimeric nucleic acid ligands having high binding affinity for first and second target molecules from a chimeric nucleic acid ligand library, said method comprising:

  • a) preparing a chimeric nucleic acid ligand library generated according to a method comprising;

    i) generating a first library of nucleic acid ligands selected through the SELEX procedure for binding to said first target molecule, said nucleic acid ligands having a 3'"'"' fixed sequence, and producing the double-stranded form of said first library of nucleic acid ligands;

    ii) generating a second library of nucleic acid ligands selected through the SELEX procedure for binding to said second target molecule, said nucleic acid ligands having a 5'"'"' fixed sequence identical to the 3'"'"' fixed sequence of the nucleic acids of said first library, and producing the double-stranded form of said second library of nucleic acid ligands;

    iii) mixing said first and second libraries under conditions which promote interlibrary annealing;

    iv) forming chimeric nucleic acid ligands by enzymatically extending the recessed 3'"'"' ends while copying the 5'"'"' extensions of each annealed pair, to yield a double-stranded chimeric nucleic acid ligand library;

    v) amplifying said double stranded nucleic acid ligand library chimeric;

    b) contacting said amplified chimeric nucleic acid ligand library with said first and second target molecules, wherein chimeric nucleic acid ligands having an increased affinity to said first and second target molecules may be partitioned from the remainder of the chimeric nucleic acid ligand library;

    c) partitioning said increased affinity chimeric nucleic acid ligands from the remainder of said chimeric nucleic acid ligand library; and

    d) amplifying said increased affinity chimeric nucleic acid ligands to yield a mixture of chimeric nucleic acid ligands enriched in members which bind to said first and second target molecules, whereby chimeric nucleic acid ligands having high binding affinity to said first and second target molecules are identified.

View all claims
  • 6 Assignments
Timeline View
Assignment View
    ×
    ×