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`Methods of synthesizing diverse coll-qctioggglolig-qmg1g*_*
`*bry|*wre4&4
`s.6flXrMqrur@r@"
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`ILMN EXHIBIT 1014
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`SiNO Ino TowNsEND
`lreet Tower
`Plua
`ilco, CA 94105
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`ER OF PATENTS AND TRADEMARKS
`tngtont D.C. 20231
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`I pttcnt apptication.
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`For:
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`Enclosed are:
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`i
`sheet(s)of I I formal [ | informaldrawing(s).
`t I An assignment of the invention
`IX A I lsigcd [{ unsigrcd Declaradon& Fowerof Attomev.
`t I Al lsigncd I lunsigredDcclaration.
`t I A power of attorney. ( unsi-gned)
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`Page 2 of 80
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`ffi,
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`Attorney Docket No. 1L509-36
`
`PATENT APPLICA,TION
`
`METHOD OF SYNTHESIZI\IG DMRSE COLLECTIONS OF OLIGOMERS
`
`Inventors:
`
`Assignee:
`
`Willian J. Dower,
`States;
`Ronal-d W. Barrett,
`States; and
`Mark A. Ga11op, a
`
`a ci.tizen of the United
`
`a citizen of the United
`
`citizen of New Zealand.
`
`Affymax Technologies N.V.
`
`TOWNSEND and TOWNSEND
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`Page 3 of 80
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`

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`q
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`qsp
`\I {.- t
`T-S'
`tgg1'{
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`ill ,t?62nZZA
`
`PATENT
`Attorney Docket No. 1"1"509-35
`
`10
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`FIELD OF THE TNVENTION
`The present invention relates generally to a general
`stochastic nethod for synthesizing random oligomers on
`particles. A further aspect of the invention relates to the
`use of identification
`tags on the particles to facilitate
`identification of the oligomer sequence.
`
`BACKGROUND OF THE INVENTION
`.
`The relationship between structure and activity of
`molecules is a fundamental issue in the study of biological
`systems. Structure-activity relationships are irnportant in
`understanding, for example, the function of enzymes, the ways
`in which cells communicate with each other, ds wel] as ceIlular
`control and feedback systems. Certain macromolecules are known
`to interact and bind to other molecules having a very specific
`three-dimensional spatial and electronic distribution.
`Any
`large molecule having such specificity can be considered a
`receptor, whether it is an enzyme catalyzing hydrolysis of a
`metabolic intermediate, a cell-surface protein nediating
`rnembrane transport of ions, a glycoprotein serving to identify
`a particular ceII to its neighbors, dn lgG-class antibody
`circurating in the plasma, drl oligonucleotide sequence of DNA
`in the nucleus, or the like. The various molecules which
`receptors selectively bind are known as ligands.
`Many assays are available for measuring the binding
`affinity of known receptors and ligands, but the inforrnation
`which can be gained from such experiments is often limited by
`the nurnber and type of ligands which are availabre. Novel
`rigands are sometines discovered by chance or by apprication of
`new techniques for the elucidation of molecular structure,
`including x-ray crystallographic analysis and reconbinAnt
`genetic technigues for proteins.
`
`
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`2
`Snal1 peptides are an exemplary systern for exploring
`the relationship between structure and function in biology. A
`peptide is a sequence of amino acids. When the twenty
`naturally occurring anino acids are condensed into polymeric
`molecules they form a wide variety of three-dirnensional
`configurations, each resulting from a particular amino acid
`seqluence and solvent condition. The number of, possible
`pentapeptides of the 20 naturally occurring amino acids, for
`example, is 2Os or 3.2 mitlion different peptides. The
`Iikelihood that molecules of this size night be useful in
`receptor-binding studies is supported by epitope analysis
`studies showing that some antibodies recognize seqluences as
`short as a few amino acids with high specificity.
`Furthermore,
`the average molecular weight of amino acids puts srnall peptides
`in the size range of many currently useful pharmaceutical
`products. Of course, larger peptides may be necessary for many
`purposes; and polypeptides having changes i-n only a sma1l
`number of residues may also be useful for such purpor"J?n"
`A
`analysis of structure-activity relationships.
`Pharmaceutical drug discovery is one type of research
`which relies on such a study of structure-activity
`relationships.
`In most cases contemporary pharmaceutical
`research can be described as the process of discovering novel
`ligands with desirable patterns of specificity
`for biologically
`important receptors. Another example is research to discover
`new compounds for use in agriculture, such as pesticides and
`herbicides.
`Prior methods of preparing large nurnbers of different
`oligomers have been painstakingly slow when used at a scale
`sufficient to pernit effective rational or random screening.
`For example, the "Merrifieldrr rnethod (J. Arn. Chen. Soc. (L963)
`85:2I49-2L54t which is incorporated herein by reference) has
`been used to synthesize peptides on a solid support. In the
`Merrifield method, an amino acid is covalently bonded to a
`support rnade of an insoluble polyner. Another amino acid with
`an alpha protected group is reacted with the covalently bonded
`anino acid to fonn a dipeptide. After washing, the protective
`group is removed and a third amino acid r.rith an alpha
`
`Ir
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`3
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`protective group is added to the dipeptide. This process is
`continued until a pept,ide of a desired rength and sequence is
`obtained. Using the Merrifield method, it is not econornically
`practical to synthesize more than a handful of peptide
`sequences in a day.
`To synthesize larger numbers of oligomer sequences,
`it has also been proposed to use a series of reaction vessels
`for oligomer synthesis. For example, a tuburar reactor system
`rnay be used to synthesize a linear oligorner on a sorid phase
`support by automated seguential addition of reagents. This
`nethod still does not enabre the synthesis of a sufficientry
`large number of origomer sequences for effective economical
`scrgening.
`
`Methods of preparing a plurality of oligomer
`sequences are also known in which a foraminous container
`encloses a known guantity of reactive solid supports, the solid
`supports being larger in size than openings of the container.
`The containers may be selectively reacted with desired
`materials to synthesize desired sequences of product molecules.
`As with other rnethods known in the art, this rnethod cannot
`practically be used to synthesize a sufficient variety of
`polypeptides for effective screening.
`Other techniques have also been described. These
`methods include the synthesis of peptides on 96 plastic pins
`which f it the forrnat of standard rnicrotiter plates.
`Unfortunately, whj-le these technigues have been somewhat
`useful, substantial problems remain. For example, these
`nethods continue to be linited
`in the diversity of seguences
`which can be economically synthesized and screened.
`Fron the above, it is seen that an improved nethod
`and apparatus for synthesizing a diverse collection of chemical
`sequences is desired.
`
`SUIIO,IARY OF THE INVENTION
`The present invention provides a general stoehastic
`nethod for synthesizing randorn oligoners on solid supportsr of
`particles. The oligomers are composed of a seguence of
`monomers, the nonomers being any member of the set of nolecules
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`which can be joined together to fonn an oligomer or poI1rmer.,
`i.e. amino acids, nucleic acids, carbohydrates, 1j-pids,
`polyesters, and the like. The nethod involves producing a
`large library of solid supports, each support having attached a
`single oligomer sequence, the oligorners being synthesized in a
`random conbinatorial (ttstochasticrr) fashion. The library is
`then screened to isolate individual solid supports carrying
`oligomers that bind to a receptor. Each oligomer seqluence in
`the library is unigue, in a preferred ernbodiment. fn another
`preferred ernbodiment, the solid supports are nonporous beads.
`The solid supports may be composed of a single particle, or two
`or more linked particles.
`A further enbodirnent of the invention is the use of
`an identifier
`tag to identify the seguence of monomers in the
`oligomer. The identifier
`tag, which rnay be attached to the
`same particle as the oligomer or to a second particle attached
`to the oligomer-carrying particle, may be any recognizable
`feature that in some way carries the required information, and
`that is decipherable at the level of one or a few solid
`supports. The solid supports may be joined to the oligomers
`and the identifier
`tag by means of a linker molecule.
`In a preferred enbodiment, the identifier
`tag will be
`an oligonucleotide, preferably composed of pyrinidines. The
`tag may contain a 5 | and a 3 |
`oligonucleotide identifier
`amplification site, to allow arnplification of the tag by, for
`example, polymerase chain reaction. A DNA seguencing prirner
`site, which may be specific for each step of the oligomer
`synthesis, may also be included in the oligonucleotide tag.
`The tag may be designed to include, in the oligonucleotide
`sequence, information allowing identification of the monomer
`associated with the addition of the particular tag. The
`oligonucleotide will be about l-00 nucleotides in length, j-n a
`preferred embodiment.
`
`BRIEF DESCRIPTTON OF THE FIGURES
`Fig. 1 is a schematic representation of cornbinatorial-
`oligorner synthesis on particles.
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`Fig. 2 is a schenatic representation of concurrent
`cornbinatorial oligomer synthesis and particle tagging'
`Fig. 3 is a description of one method of bead
`functionalization, the compatible chemistries for peptide
`synthesis and round by round attachrnent of oligonucleotide
`identifier tags, ipclusing synthesis of amino-functionalized
`b...i{,i*ild'Er-fr 5tt'q.lr!d'.g,3;gypignareinidvr
`o r iqonuc r eot ia&fuittii n8'-"dii1 Fffit iif /ana i nt roduct i on o f a
`tnior "rl.n116',1' ^'!f.?S$#tt {t Lrisonucreotide attachment to a
`u.uf i itJtbJ& ,rAfft*#G i{.ia",,+9.go,uP} ing ( s ) ?ng, oJ isonucleotide '
`attaJfi ne".r=#.iill*&:,*iifftlEirtr"""*,tiiritJ$i;ii*ti..ii''fl x*
`,,rn.fun is a schematic representation"5f one example
`of an oligonucleotiderfag-
`\2'
`DESCRIPTIoNoTTHESPECIFICEMBoDIMENTS
`Thepresentinventionprovidesnovelmethodsand
`instruments for producing large synthetic oligomer libraries'
`In a preferred embodiment of the present invention, each member
`of such a library has a means for uniguely identifying the
`sequence of each oligorner. Methods for screening such
`Iibraries and reagents useful for their production are also
`provided.
`
`n
`
`n'?
`
`j=$- ':r
`
`Glossarv
`
`intended
`are used
`
`to have the
`herein:
`
`The following tenns are
`following general meanings as they
`: Refers
`to base pairing between nucleotides or nucleic acids, such as,
`for instance, between the two strands of a double stranded DNA
`molecule or between an oligonucleotide primer and a primer
`binding site on a single stranded nucleic acid to be seguenced
`or amplified. Complementary nucleotides are' generally, A and
`T(orAandU)lofCandG.Twosing}estrandedRNAorDNA
`nolecules are said to be substantially cornplernentary when the
`nucleotides of one strand, optinally aligned and compared and
`with appropriate nucleotide j.nsertions or deletions, $air with
`at least about 80? of the nucleotides of the other strand,
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`6
`usually at least about 90t to 95*, and rnore preferably at least
`about 98 to 99.5?.
`Alternatively, substantial complementarity exists
`when an RNA or DNA strand will hybridize under selective
`5 hybridization conditions to its cornplement. Typically,
`selective hybridization will occur when there is at least about
`55t identity over a stretch of at least L4 to 25 nucleotides,
`preferably at least about 652, more preferably at least about
`752, and most preferably at least about 90t identity. See, M.
`L0 Kanehisa Nucleic Acids Res. L2z2O3 (L984), incorporated herein
`by reference.
`Stringent hybridization conditions will typically
`include salt concentrations of less than about 1 M, more
`usually less than about 5OO mM and preferably less than about
`15 200 nM. The hybridization temperature for oligomers will
`typically be greater than 22"e, more typically greater than
`about 30'C, and preferably in excess of about 37'C. Longer
`fragments may require higher hybridization temperatures for
`specific hybridization. As other factors may dramatically
`.
`20 affect the stringency of hybridization, including base
`conposition and length of the complenentary strands, presence
`of organic solvents and extent of base mismatching, the
`conbination of parameters is.more important than the absolute
`measure "'";I;:i
`fdlifi*gt fS? an antisen molecure which is
`delineated by the area of interaction with the subclass of
`rec ept o r=
`:": :::i'u1Mhl, $kS?, P#:" 95},.
`:::::, : :
`" "r i ry
`
`"
`which monomer reactions an individual solid support has
`experienced in the synthesis of an oligomer. The identifier
`tag also records the step in the synthesis series in which the
`solid support visited that monomer reaction. The identifier
`tag nay be any recognizable feature which is, for exarnple:
`microscopically distinguishable in shape, size, co1or, optical
`density, etc.; differently absorbing or emitting of light;
`chenically reactive; rnagnetically or electronically encoded; or
`in some other way distinctively marked with the required
`information, and decipherable at the leveL of one (or fe.rv)
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`7
`solid support(s). A preferred example of such an identifier
`tag is an oligonucleotide seguence.
`Ligand: A ligand is a molecule that is recognized by
`a particular receptor. The agent bound by or reacting with a
`receptor is called a rrligandrr, a term which is def initionally
`meaningful only in terms of its counterpart receptor. The term
`"ligandrr does not inply any particular molecular size or other
`structural or compositional feature other than that the
`substance in question is capable of binding or otherwise
`interacting with the receptor. A1so, a ligand may serve either
`as the naturaL ligand to which the receptor binds, or as a
`functional analogue that nay act as an agonist or antagonist.
`Exanples of ligands that can be investigated by this invention
`include, but are not restricted to, agonists and antagonists
`for bell membrane receptors, toxins and venoms, viral epitopes,
`hormones (e.9., opiates, steroids, etc,), hormone receptors,
`peptides, enzymes, enzyme substrates, cofactors, drugs,
`proteins, and monoqlonal antibodies.
`Monomer: l Sfirffil93fl.n" set or molecures which
`can be joined together to form an oligomer or pollrrner. The set
`-J\
`of monomers useful in the present invention includes, but is
`not restricted to, for the example of peptide synthesis, the
`set of L-amino acids, D-amino acids, oy synthetic amino acids.
`As used herein, monomery' reters to any rnember of a basis set
`for synthesis of an oligorner. For example, dimers of L-amino
`acids form a basis set of 400 monomers for synthesis of
`polypeptides. Different basis sets of rnonomers may be used at
`successive steps in the synthesis of a polymer.
`Oligomer or Polymer: The oligomer or polymer
`sequences of the present invention are formed from the chemical
`or enzymatic addition of nonomer subunits. Such oligomers
`include, for example, both linear, cyclic, and branched
`pollnners of nucleic acids, polysaccharides, phospholipids, and
`peptides having either c-, F- , or c,;-amino acids, heteropolymers
`in which a known drug is covalently bound to any of the above,
`polyurethanes, polyesters, polycarbonates, polyureas,
`polyanides, polyethyleneimines, polyaryJ.ene sulfides,
`polysiloxanes, polyirnides, polyacetates, oF other pollrners
`
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`I
`which will be readily apparent to one skilled in the art upon
`revi.ew of this disclosure.
`Peptide: A peptide is an oligomer in which the
`monomers are alpha amino acids and which are joined together
`through amide bonds and alternatively referred to as a
`polypeptide. In the context of this specification it should be
`appreciated that the anino acids may be the L-optical isomer or
`the D-optical isomer. Peptides are more than two amino acid
`monomers 1ong, and often more than 20 amino acid monomers long.
`Standard abbreviations for anino acids are used (e.9., P for
`proline). These abbreviations are included in Stryer,
`Biochemistrv, Third Ed., 1988, which is incorporated herei-n by
`reference.
`'
`Oliqonucleotides: An oligonucleotide is a single-
`stranded DNA or RNA molecule, typically prepared by synthetic
`neans. Those oligonucleotides ernployed in the present
`invention will be 50 to l-50 nucleotides in length, preferably
`from 80 to 120 nucleotides, and rnost preferably about l-00
`nucleotides, although oligonucleotides of different length may
`be appropriate. Suitable oligonucleotides may be prepared by
`the phosphoramidite rnethod described by Beaucage and
`Carruthers, Tetra. Letts. 22:1-859-LB6Z (L981-), or by the
`triester nethod according to Matteucc.i, et a1., J. Am. Chern.
`Soc., L03:3L85 (L981), both incorporated herein by reference,
`or by other methods such as conmercial automated
`oligonucleotide synthesizers.
`operably linked: A nucleic acid is operably linked
`when it is placed into a functional relationship with another
`nucleic acid sequence. For instance, . pr?L?:ff or enhancer is
`operably linked to a coding sequence if it*d$ffis
`the
`transcription of the sequence. Generally, operably linked
`means that the DNA sequences being linked are contiguous and,
`where necessary to join two protein coding regions, contiguous,
`and in reading f rame. sggd@'t}
`a
`Receptor: A^molecule that has an affinity
`for a
`given ligand. Receptors rnay be naturaLly-occurring or'manmade
`nolecules. AIso, they can be enployed in their unaltered
`natural or isolated state or as aggregates with other species.
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`9
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`Receptors may be attached, covalently or noncovalently, to a
`binding member, either directly or via a specific binding
`substance. Exanples of receptors which can be ernployed by this
`invention include, but are not restricted to, antibodies,
`cell rnernbrane receptors, monoclonal antibodies and antisera
`reactive with specific antigenic determinants (such as on
`viruses, cells or other naterials), drugs, polynucleotides'
`nucleic acids, peptides, cofactors, lectins' sugars'
`polysaccharides, cells, cellular rnembranes, and organelles.
`Receptors are sometirnes referred to in the art as anti-ligands'
`As the term receptors is used herein, ro difference in meaning
`is intended. A t'ligand-receptor pairn is formed when two
`macromolecules have combined through molecular recognition to
`form a comPlex.
`other examples of receptors which can be investigated
`by this invention include but are not restricted to:
`a) Microorqanism receptors: Determination of ligands that
`bind to receptors, such as specific transport proteins or
`enzvmes esSential to survival of microorganisms, is useful
`. &irco\&hl\?
`rnAa neriblass of antibiotics. Of particular value would
`be antibiotics against opportunistic fungi, protozoa' and
`those bacteria resistant to the antibiotics in current
`use '
`{gcrn ''nc\M€$
`'&tit
`b) EnzJrmes: For instance,'^tn" binding site of enzymes such
`as the enzymes ,.=porr=ibl" for cleaving neurotransmitters.
`Detennination of ligands that bind to certain receptors,
`and thus rnodulate the action of the enzymes that cleave
`the different neurotransmitters, is useful in the
`development of drugs that can be used in the treatment of
`disorders of neurotransmission.
`c) Antibodies: For instance, the invention may be useful in
`investigating the ligand-binding site on the antibody
`molecule which conbines with the epitope of an antigen of
`interest. Deterrnining a sequence that rnirnics an antigenic
`epitope rnay lead to the development of vaccines of which.
`the immunogen is based on one or more of such seqfuences,
`or lead to the developnent of related diagnostic..agents or
`compounds useful in therapeutic treatments such as for
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`d)
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`e)
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`f)
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`autoimmune diseases (e.9., by blocking the binding of the
`rrself rr antibodies) .
`Nucleic Acids: The invention may be useful in
`investigating sequences of nucleic acids acting as binding
`sites for cellular proteins (rrtrans-acting factorsrr).
`Such seqfuences may include, e.g. r enhancers or promoter
`sequences.
`{t[5 \$rt\ fckf.S So pdlrncns
`Catalvtic Polypeptides' t" rreJ+melrs, preferably
`polypeptides, which .r"u.up.bIe of promoting a chemical
`reaction involving the conversion of one or more reactants
`to one or nore products. Such polypeptides generally
`include a binding site specific for at least one reactant
`'or reaction intermediate and an active functionality
`is
`proxirnate to the binding site, which functionality
`capable of chernically nodifying the bound reactant.
`Catalytic polypeptides are described in, Lerner, R.A., et
`dI., Science 252: 659 (L991"), which is incorporated
`herein by reference.
`Nrib \gefl'tnciurdes
`Hormone recentors: For instancer^the receptors for
`insulin and growth hormone. Deterrnination of the ligands
`to a receptor is useful in
`which bind with high affinity
`the development of, for exarnple, an oral replacement of
`the daily injections which diabetics must take to relieve
`the symptoms of diabetes, and in the other case, a
`replacement for the scarce human growth hormone that can
`only be obtained from cadavers or by recombinant DNA
`technology. other examples are the vasoconstrictive
`hormone receptors; determination of those ligands that
`bucn 5
`bind to q?eceptor- rnay lead to the development of drugs to
`,.I
`control blood pressure.
`opiate receptors: Determination of ligands that bind to
`the opiate receptors in the brain is useful in the
`developrnent of less-addictive replacements for morphine
`and related druss.
`-fufj lC.f_f,n feWS .tO A
`Substrate or Solid Supportr ,.\material having a
`rigid or semi-rigid surface. Such materials will preferably
`take the form of small beads, pellets, disks or other
`,
`convenient fonns, although other forms may be used. In some
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`s)
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`ernbodiments, at least one surface of the substrate will be
`substantially flat. A roughly qpberigql rshape is preferred.
`synthet i c,'fu'A=!m.Sys t9f4ft9o "n"'i". r or
`enzymatic synthesis. The synthetic libraries of the present
`invention may be contrasted with those in viral or plasnid
`-"A
`vectors, for instance, which rnay be propagated in bacterial,
`yeastr or other living hosts.
`
`Methods for Producinq Larqe Svnthetic oligoner Libraries
`A general nethod of randon oligomer synthesis is
`provided that produces the enormous numbers of compounds
`available with recombinant systems and the monomer set
`diversity available with chemical synthesis methods. By means
`of the present method it is possible to readily produce up to
`1012 different oligorners, a drarnatic improvement over previous
`methods. It also provides a facile means of oligomer
`identification.
`The general nethod comprises:
`(a) Producing a large, highly diverse collection or
`library, each member^pf sucl,r .q l.lb.faqy.
`supports havins aEEffiWoffitftft+ffi
`peptide). The solid support is attached to the oligomer by
`neans of a linker that has an appropriate functional group at
`each end, one group appropriate for attachment to the support
`and the other group appropriate for attachment to the oligorner.
`Such a collection rnay contain, for example, all conbinations of
`n monomers assembled into X length oligorners (n* = up to l-08 to
`LO12 compounds) .
`fn one embodiment, n will be 5 or 6 t and the
`collection will contain about L05 or l-06 different nembers,
`respectively. It may also contain oligomers having different
`mononer units at, for example, only one or a small number of
`positions while having an identical seqluence at all other
`positions;
`(b) Synthesizing the oligomers in a random combinatorial
`(t'stochasticrr) fashion by chernical and/or enzymatic assembly of
`monomer unitsi
`(c) Screening the collection to isolate individual solid
`supports carrying oligorners active in binding to a receptor.
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`b)
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`A synthetic oligomer library may be produced by
`synthesizing on each of a plurality of solid supports a single
`oligomer sequence, the oligoner sequence being different for
`different solid supports. The oligomer sequence is synthesized
`in a process comprising the steps of:
`a) apportioning the supports in a stochastic manner
`among a plurality of reaction vessels;
`exposing the supports in each reaction vessel to
`a first monomer;
`c) pooling the supportsi
`d) apportioning the supports in a stochastic manner
`among the plurality of reaction vessels;
`exposing the supports in each reaction vessel to
`a second monomer; and
`repeating steps a) through e) from at least one
`f)
`to twenty tines.
`Typically, substantially egual numbers of solid
`fn one
`supports will be apportioned to each reaction vessel.
`embodirnent of the rnethod, the monomers are chosen from the set
`20 of amino acids and the resulting oligoner is a peptide.
`In a preferred ernbodiment of the invention, the solid
`supports on which the oligomers are synthesized also have
`tag that can be easily decoded to report
`attached an identifier
`the sequence of the oligorner contained on each solid support.
`25 The identifier
`tags rnay be attached to the solid support by
`means of a linker that has an appropriate functional group at
`each end, one group appropriate for attachment to the support
`and the other group appropriate for attachment to the
`identifier tag.
`It will be readily appreciated after reading
`30 the disclosure below that one could also produce large
`synthetic oligomer libraries lacking identifier
`tags.
`A synthetic oligomer library that incorporates
`identifier tags is produced by synthesizinq on each of a
`plurality of solid supports a single oligoner seqluence and one
`35 or more identifier tags identifying the oligorner sequence. The
`tags are synthesized in a
`oligomer seguence and identifier
`process conprising the steps of:
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`b)
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`c)
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`e)
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`a) apportioning the supports among a plurality of
`reaction vessels;
`exposing the supports in each reaction vesseL to
`a first oligorner monomer and to a first
`identifier tag monomer;
`pooling the supports;
`apportioning the supports among a plurality of
`reaction vessels; and
`exposing the supports to a second oligorner
`monomer and to a second identifier tag monomer.
`one embodinent, the steps of this process will be
`In
`repeated one to about 20 tirnes.
`'
`Alternatively to exposing the solid supports to a
`tag monomer at the same
`oligomer monomer and an identifier
`time, the supports rnay be exposed seguentially to the first
`tag monomer.
`identifier
`oligomer monomer and then to the first
`The supports are then pooled and exposed to the second oligorner
`tag monomer. These
`monomer and then to the second identifier
`steps are then repeated, typically from one to about 20 times.
`The invention is described herein prirnarily with
`regard to the preparation of molecules containing sequences of
`amino acids, but could readily be applied in the preparation of
`other oligomers, as can be apprecj.ated by those skilled in the
`art.
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`There are various solid supports useful in
`preparation of the synthetic oligorner libraries of the present
`invention.
`It is understood that such solid supports are solid
`phase supports commonly used for solid phase synthesis of, for
`example, such oligomers as enunerated above, and thus are well
`known to those skilled in the art.
`In some embodiments of the
`present invention, such solid supports have novel features as
`described below
`The chernical or enzymatic synthesis of the oligomer
`Iibraries of the present invention takes pLace on such solid
`supports. The term "soIid support'r as used herein embiaces a
`particle with appropriate sites for oligomer synthesis and, in
`some embodiments, tag attachnent and/or synthesis, or it. nay be
`a nore nassive solid support of up to l- mm in size.
`In
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`general, the solid support size is in the range of l- nm to
`1-00 pn. such solid supports rnay be of any shape, although they
`will preferabry be roughly spherical. such solid supports may
`consist of many materials, rimited prinariry by capacity for
`5 derivatization to attach any of a number of chemically reactive
`groups, and compatibility with the chemistry of oligomer
`synthesis and tag attachrnent. They need not necessarily be
`homogenous in size, shape, or cornposition; though usually will
`be somewhat uniform. Two or more distinctly different
`L0 populat,ions of solid supports may be used for certain purposes.
`Except as otherwise noted, the chemically reactive groups with
`which such solid supports rnay be derivatized are those cornmonly
`used"for solid state synthesis of the respective oligomer and
`thus will be well known to those skilled in the art.
`It should be noted that the solid supports of the present
`invention, as defined above, would not include such things as
`Iiving cells, viruses, or cloning vectors such as phage vectors
`or plasnids.
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`An fllustration of the Method
`As a specific example of the method, one may consider
`the synthesis on resin beads of pepticles three residues in
`length, assembled from a monomer set of three dj.fferent monomer
`reaction conponents: A, B, and c. The first monomer is
`coupled to beads, and the beads frorn all the reactions are then
`25 pooled (see, Fig. i.). The poor now contains approximatery
`equal numbers of solid