Method For Enrichment Involving Elimination By Mismatch Hybridisation
First Claim
1. A method for identifying a display molecule of a bifunctional complex further comprising an identifier oligonucleotide linked to the display molecule, said method comprising the steps ofi) separating complementary single stranded identifier oligonucleotides from an enriched and partitioned fraction of bifunctional complexes having an affinity for a target molecule, said bifunctional complexes comprising a duplex identifier oligonucleotide comprising said complementary single stranded identifier oligonucleotides,ii) hybridising the single stranded identifier oligonucleotides obtained in step i), thereby generating a composition comprising homo-duplex identifier oligonucleotides and hetero-duplex identifier oligonucleotides, wherein homo-duplex identifier oligonucleotides hybridise without generating any mis-matches between the complementary oligonucleotide strands, and wherein one or more mis-matches are present when complementary identifier oligonucleotides form hetero-duplexes,iii) separating homo-duplexes from hetero-duplexes,iv) obtaining a fraction comprising predominantly homo-duplexes,v) decoding the identifier oligonucleotides of the homo-duplexes, andvi) identifying based on the decoding in step v) the one or more display molecules identifier by the decoded identifier oligonucleotides.
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Abstract
The present invention relates to a method for selecting a molecule from a library of such molecules associated with identifier oligonucleotides, said molecule having affinity towards a target. The method involves contacting library with a target to allow for an interaction between the molecules and the target and partitioning a fraction enriched in identifier oligonucleotides of molecules interacting with the target. After an optional nucleic acid amplification of the partitioned fraction, the fraction is subjected to conditions at which hetero-duplexes and homo-duplexes are formed. The homo-duplexes are subsequently recovered and decoded to identify the identity of the molecule interacting with the target.
124 Citations
62 Claims
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1. A method for identifying a display molecule of a bifunctional complex further comprising an identifier oligonucleotide linked to the display molecule, said method comprising the steps of
i) separating complementary single stranded identifier oligonucleotides from an enriched and partitioned fraction of bifunctional complexes having an affinity for a target molecule, said bifunctional complexes comprising a duplex identifier oligonucleotide comprising said complementary single stranded identifier oligonucleotides, ii) hybridising the single stranded identifier oligonucleotides obtained in step i), thereby generating a composition comprising homo-duplex identifier oligonucleotides and hetero-duplex identifier oligonucleotides, wherein homo-duplex identifier oligonucleotides hybridise without generating any mis-matches between the complementary oligonucleotide strands, and wherein one or more mis-matches are present when complementary identifier oligonucleotides form hetero-duplexes, iii) separating homo-duplexes from hetero-duplexes, iv) obtaining a fraction comprising predominantly homo-duplexes, v) decoding the identifier oligonucleotides of the homo-duplexes, and vi) identifying based on the decoding in step v) the one or more display molecules identifier by the decoded identifier oligonucleotides.
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19-55. -55. (canceled)
Specification