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• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
  • B01J19/0046—Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
  • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
  • B01J2219/00277—Apparatus
  • B01J2219/00452—Means for the recovery of reactants or products
  • B01J2219/00454—Means for the recovery of reactants or products by chemical cleavage from the solid support
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
  • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
  • B01J2219/00277—Apparatus
  • B01J2219/00497—Features relating to the solid phase supports
  • B01J2219/005—Beads
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
  • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
  • B01J2219/00277—Apparatus
  • B01J2219/00497—Features relating to the solid phase supports
  • B01J2219/00502—Particles of irregular geometry
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
  • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
  • B01J2219/00277—Apparatus
  • B01J2219/0054—Means for coding or tagging the apparatus or the reagents
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
  • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
  • B01J2219/00277—Apparatus
  • B01J2219/0054—Means for coding or tagging the apparatus or the reagents
  • B01J2219/00545—Colours
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
  • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
  • B01J2219/00277—Apparatus
  • B01J2219/0054—Means for coding or tagging the apparatus or the reagents
  • B01J2219/00554—Physical means
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
  • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
  • B01J2219/00277—Apparatus
  • B01J2219/0054—Means for coding or tagging the apparatus or the reagents
  • B01J2219/00563—Magnetic means
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
  • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
  • B01J2219/00583—Features relative to the processes being carried out
  • B01J2219/00585—Parallel processes
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
  • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
  • B01J2219/00583—Features relative to the processes being carried out
  • B01J2219/00592—Split-and-pool, mix-and-divide processes
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
  • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
  • B01J2219/00583—Features relative to the processes being carried out
  • B01J2219/00596—Solid-phase processes
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
  • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
  • B01J2219/00718—Type of compounds synthesised
  • B01J2219/0072—Organic compounds
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
  • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
  • C07B2200/11—Compounds covalently bound to a solid support
• • C—CHEMISTRY; METALLURGY
  • C40—COMBINATORIAL TECHNOLOGY
  • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
  • C40B50/00—Methods of creating libraries, e.g. combinatorial synthesis
  • C40B50/14—Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support
• • C—CHEMISTRY; METALLURGY
  • C40—COMBINATORIAL TECHNOLOGY
  • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
  • C40B70/00—Tags or labels specially adapted for combinatorial chemistry or libraries, e.g. fluorescent tags or bar codes

Definitions

• the invention relates generally to the field of combinatorial chemistry, and in particular to encoding schemes for solid phase chemical libraries. More specifically, the invention relates to the production of constructs which have different physical characteristics to allow the constructs to be physically separated into different groups.
• chemical libraries are intentionally created collections of differing molecules which can be prepared either synthetically or biosynthetically and screened for biological activity in a variety of formats. Examples of chemical libraries include libraries of soluble molecules; libraries of compounds tethered to resin beads, silica chips or other solid supports; or recombinant peptide libraries displayed on bacteriaphage or other biological display vectors.
• solid supports are placed into individual reaction vessels and a first building block is synthesized onto each of the solid supports.
• Solid supports are then mixed together and redistributed to the reaction vessels where a second building block is synthesized onto the solid supports.
• the solid supports may once again be mixed and redistributed where another building block may be synthesized onto the solid supports. This process may be repeated as many times as necessary. Examples of mix and split techniques are described generally in U.S. Patent No. 5,503,805, the complete disclosure of which is herein incorporated by reference.
• a batch of resin supports may be divided into n fractions, coupling a single monomer amino acid to each aliquot in a separate reaction, and then thoroughly mixing all of the resin particles together. Repeating this protocol for a total of x cycles can produce a stochastic collection of up toeometric distribution. Further, to ensure representation of the majority of possible ligan n x different molecules, as governed by a hypergds, a relatively large number of beads should be employed. A typical value may be about ten times as many beads as the desired number of ligands.
• the compound may be cleaved from the solid supports and tested to determine if the compound produces the desired result. If so, the particular compound needs to be identified. However, since the solid support was mixed and split one or more times during the synthesis process, identifying the compound on the solid support can be challenging.
• solid supports are placed at discrete locations and a set of chemicals are synthesized onto each of the solid supports while they remain within their discrete locations.
• a ligand code is also coupled to each of the solid supports, and may be used to identify the chemicals synthesized onto their solid supports while at their discrete locations.
• the solid supports are then mixed and split where still another chemical is synthesized onto each of the solid supports.
• the chemical compound on each of the solid supports may then be identified by knowing the position of the solid support when the last building block was added, and by reading the ligand code to determine the chemical compounds added while the solid supports were at their discrete locations.
• the tags are used as a binary code to record the reaction history of each bead.
• the code can then be read directly from a single bead by electron capture capillary gas chromaphotography. Such a process is described generally in Michael H. J. Ohlmeyer, et al, Complex Synthetic Chemical Libraries Indexed with Molecular Tags, Proc. Natl. Acad. Sci. USA, vol. 90, pp. 10922-10926, December 1993, the complete disclosure of which is herein incorporated by reference.
• radio frequency memory tag Another encoding strategy proposes the use of a microelectronic device called a radio frequency memory tag.
• This tag may be embedded into a solid support and provided with a unique binary code.
• the chip is placed near a computer interface transceiver, which causes the chip to power up, read its identification tag, and send a code to the transceiver in an RF pulse.
• the complexity surrounding combinatorial libraries is often increased because of the requirement imposed by different chemists, which often prefer different types of assays in order to determine which ligands produce a positive result.
• the combinatorial library may need to be compatible with direct binding assays, lawn assays, solution assays, and the like.
• multiple libraries may need to be produced, which each include the same set of ligands but which are constructed using a different process so that each of the libraries is conducive with a different type of assay.
• the use of different types of assays to evaluate chemical libraries is described generally in Daniolos, A. et al. Pigment Cells Res., 3, 38-43 (1990); Jayawickreme, C.
• a chemical library comprises a plurality of constructs which are separable into physically distinct groups.
• the constructs in each group have at least one common physical characteristic which is physically distinct from the physical characteristics of the constructs in all other groups.
• the constructs may easily be separated into different groups based upon their common physical characteristics. Examples of physical characteristics which may be utilized include size, density, geometry, color, magnetization, charge, refractive index, and the like, as well as those described in U.S. Patent No. 5,708,153, the disclosure of which is herein incorporated by reference.
• the physical characteristics are classifiable into categories.
• each construct comprises a solid support to which are attached multiple components.
• Such components can include, for example, a ligand component and at least one linking component.
• the construct may also include one or more one ligand coding components.
• the constructs of each group have linking components which have been assembled in essentially the same manner.
• each group is representative of the process by which the group is testable, i.e., by knowing the manner of assembly of the linking components, one can determine the particular type of assay which should be employed to evaluate the ligand components.
• the manner of assembly of the linking components of each group is preferably unique to each group.
• the ligand component is composed of three different building blocks.
• each ligand code component is readable to determine two of the building blocks on each solid support.
• the ligand code may be employed to determine the first two building blocks which were synthesized while the solid supports were placed at discrete locations. The solid supports may then be mixed and split where the third building block is synthesized onto the solid supports.
• the third building block is known.
• the building blocks employed to produce the chemical composition may readily be determined by evaluating the ligand code component, while the manner of attaching the ligand components may be determined based on the physical characteristics of the construct.
• the invention further provides an exemplary method for producing a chemical library.
• the method utilizes a plurality of solid support groups, where each group includes multiple solid supports which have at least one common physical characteristic which is different from the physical characteristics of the solid supports of the other groups.
• the method proceeds by attaching at least one linking component to each of the solid supports.
• the manner of attachment of the linking components is unique to each group as compared to the other groups.
• a ligand is synthesized onto each solid support such that each group of solid supports receives the same ligands. In this way, a chemical library is produced where the manner of attachment of the linking components may readily be identified.
• Identification of the manner of attachment of the linking components is advantageous in that the process by which the group is testable to evaluate the ligand may be determined simply by knowing the common physical characteristic of the group to which the solid support pertains.
• one or more ligand coding components may also be attached. In such an event, a particular ligand may be identified by decoding the ligand component, typically after a tested construct has produced a positive result.
• the physical characteristics by which the testable processes may be identified include size, geometry, density, color, magnetization, charge, refractive index, and the like. As previously described, the physical characteristics may be classifiable into categories so that the physical characteristics may differ within a given category or across certain categories.
• the processes by which the groups may be tested include direct binding assays, lawn assays, solution assays, and the like.
• multiple solid supports are placed into physically discrete locations prior to the synthesizing step.
• Each of the locations is provided with at least one solid support from each of the groups.
• one or more building blocks may be synthesized onto each solid support while the solid supports are at the discrete locations.
• the ligand coding component is preferably representative of the building block(s).
• the solid supports are preferably mixed and allocated into a plurality of vessels where another building block is synthesized on each of the solid supports.
• the ligand may be identified by utilizing the ligand code to determine the initial building block(s).
• the process by which each group is testable may be determined simply by determining the common physical characteristics of each of the groups. In this way, the solid supports may be separated into groups where each group is tested using a different assay. If a positive result is obtained, the ligand may be determined by reading the ligand code.
• the invention further provides an exemplary method for evaluating a chemical library.
• a plurality of constructs are provided which are typed by at least one common physical characteristic.
• the constructs have been separated into groups based on their type. Following separation, each of the groups may be evaluated, with each group containing information which will assist in the evaluation process. For example, each group may require the performance of a different assay in order to evaluate the ligand. More specifically, the constructs of one group may have linking components which have been assembled in essentially the same manner so that the common physical characteristic of each group is representative of the process by which the group is testable.
• the physical characteristic is density.
• the constructs may be separated by sequentially placing the constructs within fluids having different densities and removing the constructs that rise to the top of each fluid.
• at least one of the physical characteristics may be size. With this construction, the constructs may be sieved or agitated to permit the larger sized constructs to rise to the top of the remaining constructs. The larger size constructs may then be picked from the collection, preferably utilizing a picking device which can distinguish based on size.
• the invention provides an exemplary chemical library system which comprises at least two different chemical libraries. Each chemical library comprises a plurality of solid supports onto which at least two building blocks are coupled.
• the solid supports of each library have a unique physical characteristic common to the library which allows the library to which each solid support belongs to be identified. In this way, the libraries may be combined to facilitate testing. When desired, the library from which a particular solid support originated may be determined simply by evaluating its physical characteristic.
• the physical characteristics of the solid supports are preferably selected from a group consisting of size, geometry, density, color, magnetization, charge, and refractive index.
• the invention further provides an exemplary method for evaluating two or more different chemical libraries.
• at least two different chemical libraries are provided, with each library comprising a plurality of solid supports onto which at least two building blocks are coupled. Further, the solid supports of each library have a unique physical characteristic common to the library.
• the two libraries are combined together to form a combined library, and the combined library is tested for positive outcomes. If any positive outcomes are observed, the library or libraries to which those solid supports belong are identified based on the physical characteristic of the solid supports.
• FIG. IA is a schematic diagram of four groups of constructs which differ in size according to the invention.
• Fig. IB illustrates four groups of constructs which differ in density according to the invention.
• Fig. IC illustrates four groups of constructs, at least some of which differ in density, shape, and size according to the invention.
• Fig. 2A is a schematic diagram of an exemplary construct according to the invention.
• Fig. 2B is a schematic diagram of an alternative construct according to the invention.
• Fig. 3 is a flow chart of an exemplary method for evaluating a chemical library according to the invention.
• Fig. 4 is a schematic diagram of a process for producing a chemical library according to the invention.
• Fig. 4A is an enlarged view of a discrete well of Fig. 4 taken along lines A-A.
• Fig. 4B is an enlarged view of a reaction vessel of Fig. 4 taken along line B-B.
• Fig. 5 is a schematic diagram illustrating the combination and subsequent evaluation of two or more different libraries according to the invention. DETAILED DESCRIPTION OF THE SPECIFIC EMBODIMENTS
• a “construct” as used herein, is a covalently bonded entity comprising, in any combination, some type of solid support, one or more linking components, one or more ligands, and optionally one or more ligand coding components.
• a "ligand coding component” is a component which may be linked to a solid support and which contains information about at least one of the chemicals used to construct a ligand which is also linked to the solid support.
• a "ligand” is a chemical reaction product of interest.
• a ligand can be part of a larger construct, where the ultimate goal will be to identify and/or cleave the ligand apart from the rest of the construct.
• linking component is a covalent bond or a molecular moiety that is suitable for linking two portions of a construct together.
• a "solid support” is one or more materials upon which combinatorial chemistry synthesis can be performed, including beads, solid surfaces, solid substrates, particles, pellets, discs, capillaries, hollow fibers, needles, solid fibers, cellulose beads, pore glass beads, silica gels, polystyrene beads optionally crosslinked with divinylbenzene, grafted copoly beads, polyacrylamide beads, latex beads, dimethylacrylamide beads, optionally cross-linked N, N'-bis-acryloyl ethylene diamine, glass particles coated with a hydrophobic polymer, fullerenes and soluble supports, such as low molecular weight, noncrosslinked polystyrene.
• the invention provides exemplary constructs having different physical characteristics which allows the constructs to be separated into different groups.
• the physical characteristics by which the groups may be separated facilitate the identification of a particular feature or aspect of the constructs within a given group.
• the common physical characteristics may be employed to identify the process by which the group is testable when screening the ligands.
• the common physical characteristic of a group may be used to identify a particular building block used in a synthesis process.
• the common physical characteristic may be employed to identify a particular library to which the construct belongs after two or more libraries have been combined.
• the constructs of the invention may be constructed to have a wide variety of physical characteristics which allow the constructs to be separated into groups having at least one common physical characteristic.
• physical characteristics which may be employed include size, density, geometry, color, magnetization, charge, refractive index, and the like.
• the physical characteristics may be categorized into types. For example, one type may be size and another type may be density.
• the invention may utilize differences within each type or across different types.
• constructs may be constructed to have various sizes so that the constructs may be separated into the different groups based on their size.
• some of the constructs may be differently sized while other constructs may have a different geometry.
• a variety of combinations of physical characteristics may be provided based on differences within a given type and/or across various types.
• Figs. 1 A-IC Examples of different physical characteristics which may be utilized with the invention are illustrated in Figs. 1 A-IC.
• the solid supports are grouped into four groups, with each group having a different size.
• a variety of techniques may then be employed to separate the constructs into different groups. For example, different size sieves may be employed to filter the different size constructs.
• the constructs may be agitated so that the larger size constructs rise to the top.
• a bead picking apparatus such as the one described in U.S. Patent
• FIG. IB An alternative scheme for separating constructs is illustrated in Fig. IB. According to this scheme, each group of constructs has a solid support with a different density as represented by reference symbols p ⁇ -p 4 . To separate the constructs into groups, the constructs may be placed into a solution having a given density such that the group with the lowest density rises to the top of the solution. These constructs may then be skimmed from the solution. The density of the solution is then increased until the group with the next largest density rises to the top of the solution. This process is then repeated until all the groups have been separated.
• Fig. IC illustrates another alternative for separating the constructs into different groups.
• the constructs are categorized into different types. For example, groups 1 and 2 are classified by density, group 3 is classified by shape, and group 4 is classified by size. Further, the constructs within groups 1 and 2 differ within the same type. More specifically, group 1 has a density that is different than group 2.
• a variety of techniques may then be employed to separate the constructs into their respective groups. For example, an optical detector may be employed to determine differences in size and shape. Differences in density may be detected using different density of solutions as previously described. Agitation techniques may also be employed as previously described.
• a typical construct 10 is shown schematically in Fig. 2 A.
• Construct 10 includes a solid support 12 to which is coupled a linking component 14, a ligand coding component 16 and a ligand component 18. Although shown with only one linking component 14, it will be appreciated that other numbers and arrangements of linking components may be provided.
• a linking component may be provided between ligand coding component 16 and ligand component 18.
• a construct is described in, for example, PCT International Application No. PCT/US97/05701, the complete disclosure of which is herein incorporated by reference.
• Another example of a construct 11 is shown in Fig. 2B and includes a solid support 12' and a pair of linking components 13 which are used to attach a ligand coding component 15 and a ligand 17.
• Such a construct is described in, for example, U.S. Patent No. 5,770,358, the complete disclosure of which is herein incorporated by reference.
• other types of constructs exist which may be utilized with the invention. For convenience of discussion, reference will be made hereinafter to construct 10.
• a different assay will need to be employed in order to screen the ligand component 18 following synthesis.
• direct binding assays, lawn assays, solution assays, and the like all employ different types and/or arrangements of linking components as described generally in Daniolos, A. et al. Pigment Cells Res., 3, 38-43 (1990); Jayawickreme, C. K., Proc. Natl. Acad. Sci. 91, 1614-1618; and Lam, Kit S., et al., "The One-Bead-One-Compound' Combinatorial Library Method", Chem.
• Such assays can include, for example, competitive receptor binding assays with radiolabeled ligands, various enzymatic assays, cell-based signal transduction assays, antibacterial assays, antiviral assays, and anticancer assays.
• the linking and ligand coding components are attached to solid support 12 prior to a synthesis process.
• solid support 12 may be provided with a unique physical characteristic which may be used to identify the manner of attachment of the linking components.
• the synthesis process proceeds to synthesize ligand component 18 to the solid support. As described in greater detail hereinafter, it is preferred to synthesize the same set of ligand components to each group of solid support so that each group may be independently evaluated.
• step 20 multiple types of constructs which each have the same set of ligands is produced in a manner similar to that just described.
• step 22 the constructs are physically separated into groups based on the types, i.e., based on the common physical characteristic that is unique to the group.
• an assay is selected for each of the groups. The type of assay may be determined based on the physical characteristic that is common to the group as shown in step 24.
• step 26 the selected assays are performed on each group of constructs to screen the ligands.
• the constructs which produce positive results may be decoded to identify the ligand.
• FIG. 4 one exemplary way to construct a chemical library which has groups of constructs that are defined by different physical characteristics will be described. However, as previously described, it will be appreciated that the separating techniques of the invention may be used with a wide variety of constructs.
• the process of Fig. 4 is somewhat related to the techniques described in PCT International Application No. PCT/US97/05701 , previously incorporated by reference, and in H. Mario Geysen, et al., Isotope or Mass Encoding of Combinatorial Libraries, Chem. & Biol. Vol. Ill, No. 8, pp. 679-688, August 1996, previously incorporated by reference.
• FIG. 4 employs the use of a plate 28 having a plurality of wells 30.
• wells 30 are arranged in ten rows and ten columns to form a total of 100 wells.
• a plurality of solid supports 32 are placed within each of the wells.
• Each of solid supports 32 preferably already includes one or more linking components and one or more ligand coding components.
• the manner of attachment of the linking components is identified by a physical characteristic of the solid support.
• well 30 includes solid supports 32 having four different densities as identified by reference numerals p ⁇ -p 4 .
• Another group of solid supports has a size which is significantly larger than the other solid supports.
• the size of solid supports 32 may vary depending on the particular application, and may range in size from about 1 ⁇ m to about 500 ⁇ m.
• Each well 30 preferably includes enough solid supports from each group to ensure that a complete set of ligands is produced on each of the groups.
• each column of wells 30 is referenced by a reference numeral Ai-Aio, and each of the ten rows is identified by a reference numeral Bi-Bio.
• Ai-Aio each column of wells 30
• Bi-Bio each of the ten rows is identified by a reference numeral Bi-Bio.
• These reference numerals are provided schematically to show the particular chemicals that are introduced into each of the wells.
• each of the solid supports will receive Ai and Bj as the first two building blocks.
• a table will preferably be maintained indicating which chemicals are provided into each of the wells.
• each reaction vessel 36 is schematically marked with a reference numeral Ci through Cio- These reference numerals indicate the third building block which is added to the associated reaction vessel 36.
• a reference numeral Ci indicates the third building block which is added to the associated reaction vessel 36.
• Fig. 4B because the number of reaction vessels 36 is one-tenth the number of wells 30, each reaction vessel will receive approximately ten times the number of solid supports 32.
• a variety of other sized matrices may be employed.
• the ending matrix may also be a ten by ten matrix.
• the solid supports are ready to be screened to determine any positive results.
• the type of screening to be performed will depend upon which group the construct belongs. Hence, prior to screening the solid supports within each of reaction vessels 36, the solid supports will be physically separated into groups based on the common physical characteristic of each group. When separating solid supports 32 into their respective groups, the association of solid supports 32 relative to their reaction vessel 36 will be maintained so that the last building block is still identifiable.
• One exemplary way to separate solid supports 32 into their different categories is first to agitate reaction vessels 36 and then utilize a bead picker to remove the larger size solid supports which will rise to the top of the remaining constructs.
• a solution may then be introduced into each of reaction vessels 36 to cause the least dense constructs to rise to the top of the fluid. These may be skimmed from the top and the density of the solution increased to allow the next of constructs to rise to the top. This process is repeated until all of the constructs are separated into groups.
• Each of the groups may then be screened using assays that are suitable for the particular group. If a positive result is observed, the ligand coding component on the solid support may be used to identify the first two building blocks, with the third building block being determined based upon the reaction vessel 36 from which it was removed. In this way, the ligand may easily be identified using techniques known in the art.
• Another application of the invention is the ability to combine two or more different libraries to make the next step in a process more efficient, while still providing the ability to identify the libraries from which any given construct originated. For example, two or more libraries may be combined prior to screening to make the screening process more efficient as illustrated in Fig. 5.
• the process begins by providing two or more different libraries, with libraries A and N being shown for convenience of illustration.
• the manner in which the libraries differ may greatly vary.
• the libraries may vary in that all of the building blocks and/or chemistries in one library are different from another library.
• the libraries may differ in that they were created at different times.
• the ligand coding assignments may differ between the libraries.
• the libraries may differ except for common subsets of ligands within the libraries.
• step 50 all of the libraries are combined and mixed.
• the entire library is then screened for positive outcomes. If one or more positive outcomes are observed, the physical characteristics of the positively tested constructs are evaluated to determine the library from which the constructs originated as shown in step 54. That particular library may then be individually tested. In this way, one initial screening may be performed with multiple libraries to reduce the time and effort employed to screen large quantities of constructs.

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