WO2001064942A1 - Protein scaffolds for antibody mimics and other binding proteins - Google Patents
Protein scaffolds for antibody mimics and other binding proteins Download PDFInfo
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- WO2001064942A1 WO2001064942A1 PCT/US2001/006414 US0106414W WO0164942A1 WO 2001064942 A1 WO2001064942 A1 WO 2001064942A1 US 0106414 W US0106414 W US 0106414W WO 0164942 A1 WO0164942 A1 WO 0164942A1
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
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- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
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- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
- C07K16/241—Tumor Necrosis Factors
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
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- C—CHEMISTRY; METALLURGY
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Definitions
- This invention relates to protein scaffolds useful, for example, for the generation of products having novel binding characteristics.
- Proteins having relatively defined three-dimensional structures may be used as reagents for the design of engineered products. These scaffolds typically contain one or more regions which are amenable to specific or random sequence variation, and such sequence randomization is often carried out to produce libraries of proteins from which desired products may be selected.
- One particular area in which such scaffolds are useful is the field of antibody design.
- a "minibody” scaffold which is related to the immunoglobulin fold, has been designed by deleting three beta strands from a heavy chain variable domain of a monoclonal antibody (Tramontano et al., J. Mol. Recognit. 7:9, 1994).
- This protein includes 61 residues and can be used to present two hypervariable loops. These two loops have been randomized and products selected for antigen binding, but thus far the framework appears to have somewhat limited utility due to solubility problems.
- the present invention provides a new family of proteins capable of evolving to bind any compound of interest.
- These proteins which make use of a fibronectin or fibronectin-like scaffold, function in a manner characteristic of natural or engineered antibodies (that is, polyclonal, monoclonal, or single-chain antibodies) and, in addition, possess structural advantages.
- the structure of these antibody mimics has been designed for optimal folding, stability, and solubility, even under conditions which normally lead to the loss of structure and function in antibodies.
- These antibody mimics may be utilized for the purpose of designing proteins which are capable of binding to virtually any compound (for example, any protein) of interest.
- the fibronectin-based molecules described herein may be used as scaffolds which are subjected to directed evolution designed to randomize one or more of the three fibronectin loops which are analogous to the complementarity-determining regions (CDRs) of an antibody variable region.
- CDRs complementarity-determining regions
- Such a directed evolution approach results in the production of antibody-like molecules with high affinities for antigens of interest.
- the scaffolds described herein may be used to display defined exposed loops (for example, loops previously randomized and selected on the basis of antigen binding) in order to direct the evolution of molecules that bind to such introduced loops.
- the present invention features a protein that includes a fibronectin type III domain having at least one randomized loop, the protein being characterized by its ability to bind to a compound that is not bound by the corresponding naturally-occurring fibronectin.
- the fibronectin type III domain is a mammalian (for example, a human) fibronectin type III domain; and the protein includes the tenth module of the fibronectin type III ( 10 Fn3) domain.
- compound binding is preferably mediated by either one, two, or three 10 Fn3 loops.
- the second loop of 10 Fn3 may be extended in length relative to the naturally-occurring module, or the 10 Fn3 may lack an integrin-binding motif.
- the integrin- binding motif may be replaced by an amino acid sequence in which a basic amino acid-neutral amino acid-acidic amino acid sequence (in the N-terminal to C-terminal direction) replaces the integrin-binding motif; one preferred sequence is serine-glycine-glutamate.
- the fibronectin type III domain-containing proteins of the invention lack disulfide bonds. Any of the fibronectin type III domain-containing proteins described herein may be formulated as part of a fusion protein (for example, a fusion protein which further includes an immunoglobulin F c domain, a complement protein, a toxin protein, or an albumin protein).
- any of the fibronectin type III domain proteins may be covalently bound to a nucleic acid (for example, an RNA), and the nucleic acid may encode the protein.
- the protein may be a multimer, or, particularly if it lacks an integrin- binding motif, it may be formulated in a physiologically-acceptable carrier.
- the present invention also features proteins that include a fibronectin type III domain having at least one mutation in a ⁇ -sheet sequence which changes the scaffold structure. Again, these proteins are characterized by their ability to bind to compounds that are not bound by the corresponding naturally-occurring fibronectin.
- any of the fibronectin scaffolds of the invention may be immobilized on a solid support (for example, a bead or chip), and these scaffolds may be arranged in any configuration on the solid support, including an array.
- the invention further features nucleic acids encoding any of the proteins of the invention.
- the nucleic acid is DNA or RNA.
- the invention also features a method for generating a protein which includes a fibronectin type III domain and which is pharmaceutically acceptable to a mammal, involving removing the integrin- binding domain of said fibronectin type III domain.
- This method may be applied to any of the fibronectin type III domain-containing proteins described above and is particularly useful for generating proteins for human therapeutic applications.
- the invention also features such fibronectin type III domain- containing proteins which lack integrin-binding domains.
- the invention features screening methods which may be used to obtain or evolve randomized fibronectin type III proteins capable of binding to compounds of interest, or to obtain or evolve compounds (for example, proteins) capable of binding to a particular protein containing a randomized fibronectin type III motif.
- the invention features screening procedures which combine these two methods, in any order, to obtain either compounds or proteins of interest.
- the first screening method useful for the isolation or identification of randomized proteins of interest, involves: (a) contacting the compound with a candidate protein, the candidate protein including a fibronectin type III domain having at least one randomized loop, the contacting being carried out under conditions that allow compound-protein complex formation; and (b) obtaining, from the complex, the protein which binds to the compound.
- the second screening method for isolating or identifying a compound which binds to a protein having a randomized fibronectin type III domain, involves: (a) contacting the protein with a candidate compound, the contacting being carried out under conditions that allow compound-protein complex formation; and (b) obtaining, from the complex, the compound which binds to the protein.
- the methods further involve either randomizing at least one loop of the fibronectin type III domain of the protein obtained in step (b) and repeating steps (a) and (b) using the further randomized protein, or modifying the compound obtained in step (b) and repeating steps (a) and (b) using the further modified compound.
- the compound is preferably a protein
- the fibronectin type III domain is preferably a mammalian (for example, a human) fibronectin type III domain.
- the protein includes the tenth module of the fibronectin type III domain ( 10 Fn3), and binding is mediated by one, two, or three 10 Fn3 loops.
- the second loop of 10 Fn3 may be extended in length relative to the naturally-occurring module, or 10 Fn3 may lack an integrin-binding motif.
- the integrin-binding motif may be replaced by an amino acid sequence in which a basic amino acid- neutral amino acid-acidic amino acid sequence (in the N-terminal to C-terminal direction) replaces the integrin-binding motif; one preferred sequence is serine- glycine-glutamate.
- the selection methods described herein may be carried out using any fibronectin type III domain-containing protein.
- the fibronectin type III domain-containing protein may lack disulfide bonds, or may be formulated as part of a fusion protein (for example, a fusion protein which further includes an immunoglobulin F c domain, a complement protein, a toxin protein, or an albumin protein).
- selections may be carried out using the fibronectin type III domain proteins covalently bound to nucleic acids (for example, RNAs or any nucleic acid which encodes the protein).
- the selections may be carried out using fibronectin domain- containing protein multimers.
- the selections involve the immobilization of the binding target on a solid support.
- solid supports include columns (for example, affinity columns, such as agarose columns) or microchips.
- the invention features diagnostic methods which employ the fibronectin scaffold proteins of the invention.
- diagnostic methods may be carried out on a sample (for example, a biological sample) to detect one analyte or to simultaneously detect many different analytes in the sample.
- the method may employ any of the scaffold molecules described herein.
- the method involves (a) contacting the sample with a protein which binds to the compound analyte and which includes a fibronectin type III domain having at least one randomized loop, the contacting being carried out under conditions that allow compound-protein complex formation; and (b) detecting the complex, and therefore the compound in the sample.
- the protein is immobilized on a solid support (for example, a chip or bead) and may be immobilized as part of an array.
- the protein may be covalently bound to a nucleic acid, preferably, a nucleic acid, such as RNA, that encodes the protein.
- the compound is often a protein, but may also be any other analyte in a sample. Detection may be accomplished by any standard technique including, without limitation, radiography, fluorescence detection, mass spectroscopy, or surface plasmon resonance.
- fibronectin type III domain is meant a domain having 7 or 8 beta strands which are distributed between two beta sheets, which themselves pack against each other to form the core of the protein, and further containing loops which connect the beta strands to each other and are solvent exposed. There are at least three such loops at each edge of the beta sheet sandwich, where the edge is the boundary of the protein perpendicular to the direction of the beta strands.
- Naturally occurring fibronectin is meant any fibronectin protein that is encoded by a living organism.
- randomized is meant including one or more amino acid alterations relative to a template sequence.
- Protein By a “protein” is meant any sequence of two or more amino acids, regardless of length, post-translation modification, or function. "Protein” and “peptide” are used interchangeably herein.
- RNA is meant a sequence of two or more covalently bonded, naturally occurring or modified ribonucleotides.
- a modified RNA included within this term is phosphorothioate RNA.
- DNA is meant a sequence of two or more covalently bonded, naturally occurring or modified deoxyribonucleotides.
- nucleic acid any two or more covalently bonded nucleotides or nucleotide analogs or derivatives. As used herein, this term includes, without limitation, DNA, RNA, and PNA.
- pharmaceutically acceptable is meant a compound or protein that may be administered to an animal (for example, a mammal) without significant adverse medical consequences.
- physiologically acceptable carrier a carrier which does not have a significant detrimental impact on the treated host and which retains the therapeutic properties of the compound with which it is administered.
- physiologically acceptable carrier is physiological saline.
- Other physiologically acceptable carriers and their formulations are known to one skilled in the art and are described, for example, in Remington's Pharmaceutical Sciences, (18 th edition), ed. A. Gennaro, 1990, Mack Publishing Company, Easton, PA, incorporated herein by reference.
- a “selecting” step provides at least a 2-fold, preferably, a 30-fold, more preferably, a 100-fold, and, most preferably, a 1000-fold enrichment of a desired molecule relative to undesired molecules in a population following the selection step.
- a selection step may be repeated any number of times, and different types of selection steps may be combined in a given approach.
- binding partner any molecule which has a specific, covalent or non-covalent affinity for a portion of a desired compound (for example, protein) of interest.
- binding partners include, without limitation, members of antigen/antibody pairs, protein/inhibitor pairs, receptor/ligand pairs (for example cell surface receptor/ligand pairs, such as hormone receptor/peptide hormone pairs), enzyme/substrate pairs (for example, kinase/substrate pairs), lectin/carbohydrate pairs, oligomeric or heterooligomeric protein aggregates, DNA binding protein/DNA binding site pairs, RNA/protein pairs, and nucleic acid duplexes, heteroduplexes, or ligated strands, as well as any molecule which is capable of forming one or more covalent or non-covalent bonds (for example, disulfide bonds) with any portion of another molecule (for example, a compound or protein).
- solid support is meant, without limitation, any column (or column material), bead, test tube, microtiter dish, solid particle (for example, agarose or sepharose), microchip (for example, silicon, silicon-glass, or gold chip), or membrane (for example, the membrane of a liposome or vesicle) to which a fibronectin scaffold or an affinity complex may be bound, either directly or indirectly (for example, through other binding partner intermediates such as other antibodies or Protein A), or in which a fibronectin scaffold or an affinity complex may be embedded (for example, through a receptor or channel).
- the present antibody mimics exhibit improved biophysical properties, such as stability under reducing conditions and solubility at high concentrations.
- these molecules may be readily expressed and folded in prokaryotic systems, such as E. coli, in eukaryotic systems, such as yeast, and in in vitro translation systems, such as the rabbit reticulocyte lysate system.
- prokaryotic systems such as E. coli
- eukaryotic systems such as yeast
- in vitro translation systems such as the rabbit reticulocyte lysate system.
- these molecules are extremely amenable to affinity maturation techniques involving multiple cycles of selection, including in vitro selection using RNA-protein fusion technology (Roberts and Szostak, Proc. Natl. Acad. Sci USA 94:12297, 1997; Szostak et al., U.S.S.N.
- FIGURE 1 is a photograph showing a comparison between the structures of antibody heavy chain variable regions from camel (dark blue) and llama (light blue), in each of two orientations.
- FIGURE 2 is a photograph showing a comparison between the structures of the camel antibody heavy chain variable region (dark blue), the llama antibody heavy chain variable region (light blue), and a fibronectin type III module number 10 ( 10 Fn3) (yellow).
- FIGURE 3 is a photograph showing a fibronectin type III module number 10 ( 10 Fn3), with the loops corresponding to the antigen-binding loops in IgG heavy chains highlighted in red.
- FIGURE 4 is a graph illustrating a sequence alignment between a fibronectin type III protein domain and related protein domains.
- FIGURE 5 is a photograph showing the structural similarities between a 10 Fn3 domain and 15 related proteins, including fibronectins, tenascins, collagens, and undulin. In this photograph, the regions are labeled as follows: constant, dark blue; conserved, light blue; neutral, white; variable, red; and RGB integrin-binding motif (variable), yellow.
- FIGURE 6 is a photograph showing space filling models of fibronectin III modules 9 and 10, in each of two different orientations. The two modules and the integrin binding loop (RGB) are labeled. In this figure, blue indicates positively charged residues, red indicates negatively charged residues, and white indicates uncharged residues.
- FIGURE 7 is a photograph showing space filling models of fibronectin III modules 7-10, in each of three different orientiations. The four modules are labeled. In this figure, blue indicates positively charged residues, red indicates negatively charged residues, and white indicates uncharged residues.
- FIGURE 8 is a photograph illustrating the formation, under different salt conditions, of RNA-protein fusions which include fibronectin type III domains.
- FIGURE 9 is a series of photographs illustrating the selection of fibronectin type III domain-containing RNA-protein fusions, as measured by PCR signal analysis.
- FIGURE 10 is a graph illustrating an increase in the percent TNF- ⁇ binding during the selections described herein, as well as a comparison between RNA-protein fusion and free protein selections.
- FIGURE 11 is a series of schematic representations showing IgG,
- FIGURE 12 is a photograph showing a molecular model of Fn-CH ! - CH 2 -CH 3 based on known three-dimensional structures of IgG (X-ray crystallography) and 10 Fn3 (NMR and X-ray crystallography).
- FIGURE 13 is a graph showing the time course of an exemplary
- FIGURES 14 and 15 are graphs illustrating TNF- ⁇ binding by TNF- ⁇ Fn-binders. In particular, these figures show mass spectra data obtained from a 10 Fn3 fusion chip and non-fusion chip, respectively.
- FIGURES 16 and 17 are the phosphorimage and fluorescence scan, respectively, of a 10 Fn3 array, illustrating TNF- ⁇ binding.
- novel antibody mimics described herein have been designed to be superior both to antibody-derived fragments and to non-antibody frameworks, for example, those frameworks described above.
- the antibody mimics described herein have no disulfide bonds, which have been reported to retard or prevent proper folding of antibody fragments under certain conditions. Since the present scaffolds do not rely on disulfides for native fold stability, they are stable under reducing conditions, unlike antibodies and their fragments which unravel upon disulfide bond breakdown.
- these fibronectin-based scaffolds provide the functional advantages of antibody molecules.
- the 10 Fn3 module is not an immunoglobulin
- its overall fold is close to that of the variable region of the IgG heavy chain ( Figure 2), making it possible to display the three fibronectin loops analogous to CDRs in relative orientations similar to those of native antibodies.
- the present antibody mimics possess antigen binding properties that are similar in nature and affinity to those of antibodies, and a loop randomization and shuffling strategy may be employed in vitro that is similar to the process of affinity maturation of antibodies in vivo.
- the antibody mimics of the present invention are based on the structure of a fibronectin module of type III (Fn3), a common domain found in mammalian blood and structural proteins. This domain occurs more than 400 times in the protein sequence database and has been estimated to occur in 2% of the proteins sequenced to date, including fibronectins, tenscin, intracellular cytoskeletal proteins, and prokaryotic enzymes (Bork and Doolittle, Proc. Natl.
- Fn3 fibronectin module of type III
- these scaffolds include, as templates, the tenth module of human Fn3 ( 10 Fn3), which comprises 94 amino acid residues.
- the overall fold of this domain is closely related to that of the smallest functional antibody fragment, the variable region of the heavy chain, which comprises the entire antigen recognition unit in camel and llama IgG ( Figure 1, 2).
- the major differences between camel and llama domains and the 10 Fn3 domain are that (i) 10 Fn3 has fewer beta strands (seven vs. nine) and (ii) the two beta sheets packed against each other are connected by a disulfide bridge in the camel and llama domains, but not in 10 Fn3.
- the three loops of 10 Fn3 corresponding to the antigen-binding loops of the IgG heavy chain ran between amino acid residues 21-31, 51-56, and 76-88 ( Figure 3).
- the length of the first and the third loop, 11 and 12 residues, respectively, fall within the range of the corresponding antigen-recognition loops found in antibody heavy chains, that is, 10-12 and 3-25 residues, respectively. Accordingly, once randomized and selected for high antigen affinity, these two loops make contacts with antigens equivalent to the contacts of the corresponding loops in antibodies.
- the second loop of 10 Fn3 is only 6 residues long, whereas the corresponding loop in antibody heavy chains ranges from 16-19 residues.
- the second loop of 10 Fn3 is preferably extended by 10-13 residues (in addition to being randomized) to obtain the greatest possible flexibility and affinity in antigen binding.
- the lengths as well as the sequences of the CDR-like loops of the antibody mimics may be randomized during in vitro or in vivo affinity maturation (as described in more detail below).
- the tenth human fibronectin type III domain, 10 Fn3, refolds rapidly even at low temperature; its backbone conformation has been recovered within 1 second at 5°C.
- One of the physiological roles of 10 Fn3 is as a subunit of fibronectin, a glycoprotein that exists in a soluble form in body fluids and in an insoluble form in the extracellular matrix (Dickinson et al., J. Mol. Biol. 236:1079, 1994).
- a fibronectin monomer of 220-250 kD contains 12 type I modules, two type II modules, and 17 fibronectin type III modules (Potts and Campbell, Curr. Opin.Cell Biol. 6:648, 1994). Different type III modules are involved in the binding of fibronectin to integrins, heparin, and chondroitin sulfate. 10 Fn3 was found to mediate cell adhesion through an integrin-binding Arg-Gly-Asp (RGD) motif on one of its exposed loops.
- RGD integrin-binding Arg-Gly-Asp
- the 10 Fn3 framework possesses exposed loop sequences tolerant of randomization, facilitating the generation of diverse pools of antibody mimics. This determination was made by examining the flexibility of the 10 Fn3 sequence.
- the human 10 Fn3 sequence was aligned with the sequences of fibronectins from other sources as well as sequences of related proteins ( Figure 4), and the results of this alignment were mapped onto the three-dimensional structure of the human 10 Fn3 domain ( Figure 5).
- mutations change the scaffold and thereby indirectly alter loop structure(s). If this approach is taken, mutations should not saturate the sequence, but rather few mutations should be introduced. Preferably, no more than 10 amino acid changes, and, more preferably, no more than 3 amino acid changes should be introduced to the ⁇ -sheet sequences by this approach.
- the antibody mimics described herein may be fused to other protein domains.
- these mimics may be integrated with the human immune response by fusing the constant region of an IgG (F c ) with a 10 Fn3 module, preferably through the C-terminus of 10 Fn3.
- the F c in such a 10 Fn3-F c fusion molecule activates the complement component of the immune response and increases the therapeutic value of the antibody mimic.
- a fusion between 10 Fn3 and a complement protein, such as Clq may be used to target cells, and a fusion between 10 Fn3 and a toxin may be used to specifically destroy cells that carry a particular antigen.
- 10 Fn3 in any form may be fused with albumin to increase its half -life in the bloodstream and its tissue penetration.
- Any of these fusions may be generated by standard techniques, for example, by expression of the fusion protein from a recombinant fusion gene constructed using publically available gene sequences.
- any of the fibronectin constructs described herein may be generated as dimers or multimers of 10 Fn3 -based antibody mimics as a means to increase the valency and thus the avidity of antigen binding.
- Such multimers may be generated through covalent binding between individual 10 Fn3 modules, for example, by imitating the natural 8 Fn3- 9 Fn3- 10 Fn3 C-to-N-terminus binding or by imitating antibody dimers that are held together through their constant regions.
- a 10 Fn3-Fc construct may be exploited to design dimers of the general scheme of
- the bonds engineered into the Fc::Fc interface may be covalent or non-covalent.
- dimerizing or multimerizing partners other than Fc can be used in 10 Fn3 hybrids to create such higher order structures.
- covalently bonded multimers may be generated by constructing fusion genes that encode the multimer or, alternatively, by engineering codons for cysteine residues into monomer sequences and allowing disulfide bond formation to occur between the expression products.
- Non-covalently bonded multimers may also be generated by a variety of techniques.
- 10 Fn3 represents a preferred scaffold for the generation of antibody mimics
- other molecules may be substituted for 10 Fn3 in the molecules described herein.
- These include, without limitation, human fibronectin modules 1 Fn3- 9 Fn3 and n Fn3- 17 Fn3 as well as related Fn3 modules from non-human animals and prokaryotes.
- Fn3 modules from other proteins with sequence homology to 10 Fn3, such as tenascins and undulins may also be used.
- Modules from different organisms and parent proteins may be most appropriate for different applications; for example, in designing an antibody mimic, it may be most desirable to generate that protein from a fibronectin or fibronectin-like molecule native to the organism for which a therapeutic or diagnostic molecule is intended.
- the antibody mimics described herein may be used in any technique for evolving new or improved binding proteins.
- the target of binding is immobilized on a solid support, such as a column resin or microtiter plate well, and the target contacted with a library of candidate scaffold-based binding proteins.
- a library may consist of 10 Fn3 clones constructed from the wild type 10 Fn3 scaffold through randomization of the sequence and/or the length of the 10 Fn3 CDR-like loops.
- this library may be an RNA-protein fusion library generated, for example, by the techniques described in Szostak et al., U.S.S.N.
- it may be a DNA-protein library (for example, as described in Lohse, DNA-Protein Fusions and Uses Thereof, U.S.S.N. 60/110,549, U.S.S.N. 09/459,190, and US 99/28472).
- the fusion library is incubated with the immobilized target, the support is washed to remove non-specific binders, and the tightest binders are eluted under very stringent conditions and subjected to PCR to recover the sequence information or to create a new library of binders which may be used to repeat the selection process, with or without further mutagenesis of the sequence. A number of rounds of selection may be performed until binders of sufficient affinity for the antigen are obtained.
- the 10 Fn3 scaffold may be used as the selection target.
- RNA-protein fusion library based on an appropriate scaffold is then allowed to interact with the support, which is then washed, and desired molecules eluted and re-selected as described above.
- the 10 Fn3 scaffold may be used to find natural proteins that interact with the peptide sequence displayed in a 10 Fn3 loop.
- the 10 Fn3 protein is immobilized as described above, and an RNA-protein fusion library is screened for binders to the displayed loop.
- the binders are enriched through multiple rounds of selection and identified by DNA sequencing.
- RNA-protein libraries represent exemplary libraries for directed evolution
- any type of scaffold-based library may be used in the selection methods of the invention.
- the antibody mimics described herein may be evolved to bind any antigen of interest. These proteins have thermodynamic properties superior to those of natural antibodies and can be evolved rapidly in vitro. Accordingly, these antibody mimics may be employed in place of antibodies in all areas in which antibodies are used, including in the research, therapeutic, and diagnostic fields. In addition, because these scaffolds possess solubility and stability properties superior to antibodies, the antibody mimics described herein may also be used under conditions which would destroy or inactivate antibody molecules. Finally, because the scaffolds of the present invention may be evolved to bind virtually any compound, these molecules provide completely novel binding proteins which also find use in the research, diagnostic, and therapeutic areas.
- a complex library was constructed from three fragments, each of which contained one randomized area corresponding to a CDR-like loop.
- the fragments were named BC, DE, and FG, based on the names of the CDR-H-like loops contained within them; in addition to 10 Fn3 and a randomized sequence, each of the fragments contained stretches encoding an N-terminal His 6 domain or a C-terminal FLAG peptide tag.
- each DNA fragment contained recognition sequences for the Earl Type IIS restriction endonuclease. This restriction enzyme allowed the splicing together of adjacent fragments while removing all foreign, non- 10 Fn3, sequences.
- each fragment was assembled from two overlapping oligonucleotides, which were first annealed, then extended to form the double-stranded DNA form of the fragment.
- the oligonucleotides that were used to construct and process the three fragments are listed below; the "Top” and “Bottom” species for each fragment are the oligonucleotides that contained the entire 10 Fn3 encoding sequence. In these oligonucleotides designations, "N” indicates A, T, C, or G; and “S” indicates C or G.
- HFnLFGTop 5'- GG AAT TCC TAA TAC GAC TCA CTA TAG GGA CAA TTA CTA TTT ACA ATT ACA ATG CAT CAC CAT CAC CAT CAC CTC TTC TAT ACC ATC ACT GTG TAT GCT GTC-3' (SEQ ID NO: 8)
- T7Tmv (introduces T7 promoter and TMV untranslated region needed for in vitro translation): 5'- GCG TAA TAC GAC TCA CTA TAG GGA CAA TTA CTA TTT ACA ATT ACA-3' (SEQ ID NO: 11)
- Unispl-s spint oligonucleotide used to ligate mRNA to the puromycin-containing linker, described by Roberts et al, 1997, supra: 5'-TTTTTTTTTNAGCGGATGC-3' (SEQ ID NO: 13)
- each of the double-stranded fragments was transformed into a RNA-protein fusion (PROfusionTM) using the technique developed by Szostak et al., U.S.S.N. 09/007,005 and U.S.S.N. 09/247,190; Szostak et al,
- the resulting mRNA-DNA-puromycin-protein PROfusionTM was purified using Oligo(dT) cellulose, and a complementary DNA strand was synthesized using reverse transcriptase and the RT primers described above (Unisplint-S or flag AS A), following the manufacturer's instructions.
- the PROfusionTM obtained for each fragment was next purified on the resin appropriate to its peptide purification tag, i.e., on Ni-NTA agarose for the His 6 -tag and M2 agarose for the FLAG-tag, following the procedure recommended by the manufacturer.
- the DNA component of the tag-binding PROfusionsTM was amplified by PCR using Pharmacia Ready-to-Go PCR Beads, 10 pmol of 5' and 3' PCR primers, and the following PCR program (Pharmacia, Piscataway, NJ): Step 1: 95°C for 3 minutes; Step 2: 95°C for 30 seconds, 58/62°C for 30 seconds, 72°C for 1 minute, 20/25/30 cycles, as required; Step 3: 72°C for 5 minutes; Step 4: 4°C until end.
- the resulting DNA was cleaved by 5 U Earl (New England Biolabs) perl ug DNA; the reaction took place in T4 DNA Ligase Buffer (New England Biolabs) at 37°C, for 1 hour, and was followed by an incubation at 70°C for 15 minutes to inactivate Ear I. Equal amounts of the BC, DE, and FG fragments were combined and ligated to form a full-length 10 Fn3 gene with randomized loops. The ligation required 10 U of fresh Earl (New England Biolabs) and 20 U of T4 DNA Ligase (Promega, Madison, WI), and took 1 hour at 37°C.
- Three different libraries were made in the manner described above. Each contained the form of the FG loop with 10 randomized residues.
- the BC and the DE loops of the first library bore the wild type 10 Fn3 sequence; a BC loop with 7 randomized residues and a wild type DE loop made up the second library; and a BC loop with 7 randomized residues and a DE loop with 4 randomized residues made up the third library.
- the complexity of the FG loop in each of these three libraries was 10 13 ; the further two randomized loops provided the potential for a complexity too large to be sampled in a laboratory.
- the three libraries constructed were combined into one master library in order to simplify the selection process; target binding itself was expected to select the most suitable library for a particular challenge.
- PROfusionsTM were obtained from the master library following the general procedure described in Szostak et al., U.S.S.N. 09/007,005 and 09/247,190; Szostak et al., WO98/31700; and Roberts & Szostak, Proc. Natl. Acad. Sci. USA (1997) vol. 94, p. 12297-12302 ( Figure 8).
- the master library in the PROfusionTM form was subjected to selection for binding to TNF- ⁇ .
- Two protocols were employed: one in which the target was immobilized on an agarose column and one in which the target was immobilized on a BIACORE chip.
- an extensive optimization of conditions to minimize background binders to the agarose column yielded the favorable buffer conditions of 50 mM HEPES pH 7.4, 0.02% Triton, 100 ⁇ g/ml Sheared Salmon Sperm DNA.
- the non-specific binding of the 10 Fn3 RNA fusion to TNF- ⁇ Sepharose was 0.3%.
- the non-specific binding background of the 10 Fn3 RNA-DNA to TNF- ⁇ Sepharose was found to be 0.1%.
- ProfusionTM library was first preincubated for an hour with underivatized Sepharose to remove any remaining non-specific binders; the flow-through from this pre-clearing was incubated for another hour with TNF- ⁇ Sepharose. The TNF- ⁇ Sepharose was washed for 3-30 minutes.
- Arginine 78 - Glycine 79 - Aspartate 80 (the "RGD motif) at the tip of the FG loop.
- SGE mutant Serine 78 - Glycine 79 - Glutamate 80
- This SGE mutant was expressed as an N-terminally His 6 -tagged, free protein in R coli, and purified to homogeneity on a metal chelate column followed by a size exclusion column.
- the DNA sequence encoding His 6 - 10 Fn3(SGE) was cloned into the pET9a expression vector and transformed into BL21 DE3 pLysS cells.
- the induced culture was further incubated, under the same conditions, overnight (14-18 hours); the bacteria were recovered by standard, low speed centrifugation.
- the cell pellet was resuspended in 1/50 of the original culture volume of lysis buffer (50 mM Tris 8.0, 0.5 M NaCl, 5% glycerol, 0.05% Triton X-100, and 1 mM PMSF), and the cells were lysed by passing the resulting paste through a Microfluidics Corporation Microfluidizer M110-EH, three times.
- the lysate was clarified by centrifugation, and the supernatant was filtered through a 0.45 ⁇ m filter followed by filtration through a 0.2 ⁇ m filter.
- the column was run at 0.8 mL/min, in buffer 1; all fractions that contained a protein of the expected MW were pooled, concentrated 10X as described above, then dialyzed into PBS. Toxikon (MA) was engaged to perform endotoxin screens and animal studies on the resulting sample.
- This 10 Fn3-F c molecule contains the -CH r CH 2 -CH 3 ( Figure 11) or -CH 2 -CH 3 domains of the IgG constant region of the host; in these constructs, the 10 Fn3 domain is grafted onto the N-terminus in place of the IgG V H domain ( Figures 11 and 12).
- Such antibody-like constructs are expected to improve the pharmacokinetics of the protein as well as its ability to harness the natural immune response.
- the -CH 1 -CH 2 -CH 3 region was first amplified from a mouse liver spleen cDNA library (Clontech), then ligated into the pET25b vector.
- the primers used in the cloning were 5' Fc Nest and 3' 5 Fc Nest, and the primers used to graft the appropriate restriction sites onto the ends of the recovered insert were 5' Fc HIII and 3' Fc Nhe:
- PCR is used to remove the CH j region from this clone and create the Fc part of the shorter, 10 Fn3-CH 2 -CH 3 clone.
- the sequence encoding 10 Fn3 is spliced onto the 5' end of each clone; either the wild type 10 Fn3 cloned from the same mouse spleen cDNA library or a modified 10 Fn3 obtained by mutagenesis or randomization of the molecules can be used.
- the oligonucleotides used in the cloning of murine wild-type 10 Fn3 were:
- the human equivalents of the clones are constructed using the same strategy with human oligonucleotide sequences.
- the suitability of the 10 Fn3 scaffold for protein chip applications is the consequence of (1) its ability to support many binding functions which can be selected rapidly on the bench or in an automated setup, and (2) its superior biophysical properties.
- the versatile binding properties of 10 Fn3 are a function of the loops displayed by the Fn3 immunoglobulin-like, beta sandwich fold. As discussed above, these loops are similar to the complementarity determining regions of antibody variable domains and can cooperate in a way similar to those antibody loops in order to bind antigens.
- 10 Fn3 loops BC (residues 21-30), DE (residues 51-56), and FG (residues 76-87) are randomized either in sequence, in length, or in both sequence and length in order to generate diverse libraries of mRNA- I0 Fn3 fusions.
- the binders in such libraries are then enriched based on their affinity for an immobilized or tagged target, until a small population of high affinity binders are generated.
- error-prone PCR and recombination can be employed to facilitate affinity maturation of selected binders. Due to the rapid and efficient selection and affinity maturation protocols, binders to a large number of targets can be selected in a short time.
- the 10 Fn3 domain As a scaffold for binders to be immobilized on protein chips, the 10 Fn3 domain has the advantage over antibody fragments and single-chain antibodies of being smaller and easier to handle.
- 10 Fn3 unlike single-chain scaffolds or isolated variable domains of antibodies, which vary widely in their stability and solubility, and which require an oxidizing environment to preserve their structurally essential disulfide bonds, 10 Fn3 is extremely stable, with a melting temperature of 110°C, and solubility at a concentration > 16 mg/mL.
- the 10 Fn3 scaffold also contains no disulfides or free cysteines; consequently, it is insensitive to the redox potential of its environment.
- a further advantage of 10 Fn3 is that its antigen-binding loops and N-terminus are on the edge of the beta-sandwich opposite to the C-terminus; thus the attachment of a 10 Fn3 scaffold to a chip by its C-terminus aligns the antigen-binding loops, allowing for their greatest accessibility to the solution being assayed. Since 10 Fn3 is a single domain of only 94 amino acid residues, it is also possible to immobilize it onto a chip surface at a higher density than is used for single-chain antibodies, with their approximately 250 residues.
- the hydrophilicity of the 10 Fn3 scaffold which is reflected in the high solubility of this domain, leads to a lower than average background binding of 10 Fn3 to a chip surface.
- the stability of the 10 Fn3 scaffold as well as its suitability for library formation and selection of binders are likely to be shared by the large, Fn3-like class of protein domains with an immunoglobulin-like fold, such as the domains of tenascin, N-cadherin, E-cadherin, ICAM, titin, GCSF-R, cytokine receptor, glycosidase inhibitor, and antibiotic chromoprotein.
- the key features shared by all such domains are a stable framework provided by two beta-sheets, which are packed against each other and which are connected by at least three solvent-accessible loops per edge of the sheet; such loops can be randomized to generate a library of potential binders without disrupting the structure of the framework (as described above).
- Fn-binders may be immobilized as RNA-protein fusions by Watson-Crick hybridization of the RNA moiety of the fusion to a base complementary DNA immobilized on the chip surface (as described, for example, in Addressable Protein Arrays, U.S.S.N. 60/080,686; U.S.S.N. 09/282,734; and WO 99/51773).
- Fn-binders can be immobilized as free proteins directly on a chip surface. Manual as well as robotic devices may be used for deposition of the Fn-binders on the chip surface.
- Spotting robots can be used for deposition of Fn-binders with high density in an array format (for example, by the method of Lueking et al., Anal Biochem. 1999 May 15;270(1): 103-11). Different methods may also be utilized for anchoring the Fn-binder on the chip surface. A number of standard immobilization procedures may be used including those described in Methods in Enzymology (K. Mosbach and B. Danielsson, eds.), vols. 135 and 136, Academic Press, Orlando, Florida, 1987; Nilsson et al., Protein Expr. Purif. 1997 Oct; 11(1):1-16; and references therein.
- Oriented immobilization of Fn-binders can help to increase the binding capacity of chip-bound Fn-binders.
- Exemplary approaches for achieving oriented coupling are described in Lu et al., The Analyst (1996), vol. 121, p. 29R-32R; and Turkova, J Chromatogr B Biomed Sci App. 1999 Feb 5;722(l-2): 11-31.
- any of the methods described herein for anchoring Fn-binders to chip surfaces can also be applied to the immobilization of Fn-binders on beads, or other supports.
- Selected populations of Fn-binders may be used for detection and/or quantitation of analyte targets, for example, in samples such as biological samples.
- samples such as biological samples.
- selected Fn-binders to targets of interest are immobilized on an appropriate support to form multi-featured protein chips.
- a sample is applied to the chip, and the components of the sample that associate with the Fn-binders are identified based on the target- specificity of the immobilized binders.
- one or more components may be simultaneously identified or quantitated in a sample (for example, as a means to carry out sample profiling).
- Methods for target detection allow measuring the levels of bound protein targets and include, without limitation, radiography, fluorescence scanning, mass spectroscopy (MS), and surface plasmon resonance (SPR).
- Autoradiography using a phosphorimager system (Molecular Dynamics, Sunnyvale, CA) can be used for detection and quantification of target protein which has been radioactively labeled, e.g., using 35 S methionine.
- Fluorescence scanning using a laser scanner may be used for detection and quantification of fluorescently labeled targets.
- fluorescence scanning may be used for the detection of fluorescently labeled ligands which themselves bind to the target protein (e.g., fluorescently labeled target-specific antibodies or fluorescently labeled streptavidin binding to target-biotin, as described below).
- fluorescently labeled ligands which themselves bind to the target protein (e.g., fluorescently labeled target-specific antibodies or fluorescently labeled streptavidin binding to target-biotin, as described below).
- Mass spectroscopy can be used to detect and identify bound targets based on their molecular mass. Desorption of bound target protein can be achieved with laser assistance directly from the chip surface as described below. Mass detection also allows determinations, based on molecular mass, of target modifications including post-translational modifications like phosophorylation or glycosylation. Surface plasmon resonance can be used for quantification of bound protein targets where the Fn-binder(s) are immobilized on a suitable gold-surface (for example, as obtained from Biacore, Sweden).
- Fn binders in this case, Fn-binders specific for the protein, TNF- ⁇
- TNF- ⁇ Fn-binders specific for the protein
- an 10 Fn3-based selection was performed against TNF- ⁇ , using a library of human 10 Fn3 variants with randomized loops BC, DE, and FG.
- the library was constructed from three DNA fragments, each of which contained nucleotide sequences that encoded approximately one third of human 10 Fn3, including one of the randomized loops.
- the DNA sequences that encoded the loop residues listed above were rebuilt by oligonucleotide synthesis, so that the codons for the residues of interest were replaced by (NNS)n, where N represents any of the four deoxyribonucleotides (A, C, G, or T), and S represents either C or G.
- N represents any of the four deoxyribonucleotides (A, C, G, or T)
- S represents either C or G.
- the C-terminus of each fragment contained the sequence for the FLAG purification tag.
- each DNA fragment was transcribed, ligated to a puromycin-containing DNA linker, and translated in vitro, as described by Szostak et al. (Roberts and Szostak, Proc. Natl. Acad. Sci USA 94:12297, 1997; Szostak et al., U.S.S.N. 09/007,005 and U.S.S.N. 09/247,190; Szostak et al., WO98/31700), to generate an mRNA-peptide fusion, which was then reverse-transcribed into a DNA-mRNA-peptide fusion.
- the binding of the FLAG-tagged peptide to M2 agarose separated full-length fusion molecules from those containing frameshifts or superfluous stop codons; the DNA associated with the purified full-length fusion was amplified by PCR, then the three DNA fragments were cut by Ear I restriction endonuclease and ligated to form the full length template.
- the template was transcribed, ligated to puromycin-containing DNA linkers, and translated to generate a 10 Fn3-PROfusionTM library, which was then reverse-transcribed to yield the DNA-mRNA-peptide fusion library which was subsequently used in the selection.
- TNF- ⁇ binders were placed in 50 mM HEPES, pH 7.4, 0.02% Triton-X, 0.1 mg/mL salmon sperm DNA.
- the PROfusionTM library was incubated with Sepharose-immobilized TNF- ⁇ ; after washing, the DNA associated with the tightest binders was eluted with 0.1 M KOH, amplified by PCR, and transcribed, ligated, translated, and reverse-transcribed into the starting material for the next round of selection.
- an oligonucleotide capture probe was prepared with an automated DNA synthesizer (PE BioSystems Expedite 8909) using the solid-support phosphoramidite approach. All reagents were obtained from Glen Research. Synthesis was initiated with a solid support containing a disulfide bond to eventually provide a 3'-terminal thiol functionality. The first four monomers to be added were hexaethylene oxide units, followed by 20 T monomers. The 5 '-terminal DMT group was not removed.
- the capture probe was cleaved from the solid support and deprotected with ammonium hydroxide, concentrated to dryness in a vacuum centrifuge, and purified by reverse-phase HPLC using an acetonitrile gradient in triethylammonium acetate buffer. Appropriate fractions from the HPLC were collected, evaporated to dryness in a vacuum centrifuge, and the 5'-terminal DMT group was removed by treatment with 80% AcOH for 30 minutes. The acid was removed by evaporation, and the oligonucleotide was then treated with 100 mM DTT for 30 minutes to cleave the disulfide bond. DTT was removed by repeated extraction with EtOAc. The oligonucleotide was ethanol precipitated from the remaining aqueous layer and checked for purity by reverse-phase HPLC.
- the 3'-thiol capture probe was adjusted to 250 ⁇ M in degassed IX PBS buffer and applied as a single droplet (75 ⁇ L) to a 9x9mm gold-coated chip (Biacore) in an argon-flushed chamber containing a small amount of water. After 18 hours at room temperature, the capture probe solution was removed, and the functionalized chip was washed with 50 mL IX PBS buffer (2x for 15 minutes each) with gentle agitation, and then rinsed with 50 mL water (2x for 15 minutes each) in the same fashion. Remaining liquid was carefully removed and the functionalized chips were either used immediately or stored at 4°C under argon.
- Figures 14 and 15 show the mass spectra from the 10 Fn3 fusion chip and the non-fusion chip, respectively. In each case, a small number of 200 micron features were analyzed to collect the spectra, but Figure 15 required significantly more acquisitions.
- the signal at 17.5 kDa corresponds to TNF- ⁇ monomer.
- Pre-cleaned 1x3 inch glass microscope slides (Goldseal, #3010) were treated with Nanostrip (Cyantek) for 15 minutes, 10% aqueous NaOH at 70°C for 3 minutes, and 1% aqueous HC1 for 1 minute, thoroughly rinsing with deionized water after each reagent.
- the slides were then dried in a vacuum desiccator over anhydrous calcium sulfate for several hours.
- a 1% solution of aminopropytrimethoxy silane in 95% acetone / 5% water was prepared and allowed to hydrolyze for 20 minutes.
- the glass slides were immersed in the hydrolyzed silane solution for 5 minutes with gentle agitation.
- silane was removed by subjecting the slides to ten 5-minute washes, using fresh portions of 95% acetone / 5% water for each wash, with gentle agitation. The slides were then cured by heating at 110°C for 20 minutes. The silane treated slides were immersed in a freshly prepared 0.2% solution of phenylene 1,4-diisothiocyanate in 90% DMF / 10% pyridine for two hours, with gentle agitation. The slides were washed sequentially with 90% DMF / 10% pyridine, methanol, and acetone. After air drying, the functionalized slides were stored at 0°C in a vacuum desiccator over anhydrous calcium sulfate. Similar results were obtained with commercial amine-reactive slides (3-D Link, Surmodics). Oligonucleotide capture probes were prepared with an automated
- DNA synthesizer (PE BioSystems Expedite 8909) using conventional phosphoramidite chemistry. All reagents were from Glen Research. Synthesis was initiated with a solid support bearing an orthogonally protected amino functionality, whereby the 3'-terminal amine is not unmasked until final deprotection step. The first four monomers to be added were hexaethylene oxide units, followed by the standard A, G, C and T monomers. All capture oligo sequences were cleaved from the solid support and deprotected with ammonium hydroxide, concentrated to dyrness, precipitated in ethanol, and purified by reverse-phase HPLC using an acetonitrile gradient in triethylammonium acetate buffer. Appropriate fractions from the HPLC were collected, evaporated to dryness in a vacuum centrifuge, and then coevaporated with a portion of water.
- the purified, amine-labeled capture oligos were adjusted to a concentration of 250 ⁇ M in 50 mM sodium carbonate buffer (pH 9.0) containing 10% glycerol.
- the probes were spotted onto the amine-reactive glass surface at defined positions in a 5x5x6 array pattern with a 3-axis robot (MicroGrid, BioRobotics).
- a 16-pin tool was used to transfer the liquid from 384- well microtiter plates, producing 200 micron features with a 600 micron pitch. Each sub-grid of 24 features represents a single capture probe (i.e., 24 duplicate spots).
- the arrays were incubated at room temperature in a moisture-saturated environment for 12-18 hours.
- the attachment reaction was terminated by immersing the chips in 2% aqueous ammonium hydroxide for five minutes with gentle agitation, followed by rinsing with distilled water (3X for 5 minutes each).
- the array was finally soaked in 10X PBS solution for 30 minutes at room temperature, and then rinsed again for 5 minutes in distilled water.
- CP3' TGTAAATAGTAATTGTCCC (SEQ ID NO: 22)
- CP5' ⁇ (SE Q ID N0 . 23)
- CPnegl CCTGTAGGTGTCCAT (SEQ ID NO: 24)
- CPflag CATCGTCCTTGTAGTC (SEQ ID NO: 25)
- CPneg2 CGTCGTAGGGGTA (SEQ ID NO: 26)
- CPneg3 CAGGTCTTCTTCAGAGA (SEQ ID NO: 27)
- Recombinant human TNF- ⁇ (500 ⁇ g, lyophilized, from PreproTech) was taken up in 230 ⁇ L IX PBS and dialyzed against 700 mL stirred IX PBS at 4°C for 18 hours in a Microdialyzer unit (3,500 MWCO, Pierce).
- the dialyzed TNF- ⁇ was treated with EZ-Link NHS-LC-LC biotinylation reagent (20 ⁇ g, Pierce) for 2 hours at 0°C, and again dialyzed against 700 mL stirred IX PBS at 4°C for 18 hours in a Microdialyzer unit (3,500 MWCO, Pierce).
- the resulting conjugate was analyzed by MALDI-TOF mass spectrometry and was found to be almost completely functionalized with a single biotin moiety.
- Each of the following processes was conducted at 4°C with continuous rotation or mixing.
- the protein microarray surface was passivated by treatment with IX TBS containing 0.02% Tween-20 and 0.2% BSA (200 ⁇ L) for 60 minutes.
- Biotinylated TNF- ⁇ 100 nM concentration made up in the passivation buffer
- the microarray was washed with IX TBS containing 0.02% Tween-20 (3X 50 mL, 5 minutes each wash).
- Fluorescently labeled streptavidin (2.5 ⁇ g/mL Alexa 546-streptavidin conjugate from Molecular Probes, made up in the passivation buffer) was contacted with the microarray for 60 minutes.
- the microarray was washed with IX TBS containing 0.02% Tween-20 (2X 50 mL, 5 minutes each wash) followed by a 3 minute rinse with IX TBS. Traces of liquid were removed by centrifugation, and the slide was allowed to air-dry at room temperature. Fluorescence laser scanning was performed with a GSI Lumonics ScanArray 5000 system using 10 ⁇ M pixel resolution and preset excitation and emission wavelengths for Alexa 546 dye.
- Phosphorimage analysis was performed with a Molecular Dynamics Storm system. Exposure time was 48 hours with direct contact between the microarray and the phosphor storage screen. Phosphorimage scanning was performed at the 50 ⁇ M resolution setting, and data was extracted with ImageQuant v.4.3 software.
- Figures 16 and 17 are the phosphorimage and fluorescence scan, respectively, of the same array.
- the phosphorimage shows where the 10 Fn3 fusion hybridized based on the 35 S methionine signal.
- the fluorescence scan shows where the labeled TNF- ⁇ bound.
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---|---|---|---|---|
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US7115396B2 (en) | 1998-12-10 | 2006-10-03 | Compound Therapeutics, Inc. | Protein scaffolds for antibody mimics and other binding proteins |
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Families Citing this family (311)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
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GB0417887D0 (en) | 2004-08-11 | 2004-09-15 | Ares Trading Sa | Protein |
GB0423126D0 (en) * | 2004-10-18 | 2004-11-17 | Ares Trading Sa | Protein |
GB0426960D0 (en) * | 2004-12-08 | 2005-01-12 | Ares Trading Sa | TGR-3 like protein receptor |
US20080254451A1 (en) * | 2004-12-28 | 2008-10-16 | Ares Trading S.A. | Compositions and Methods for Treating Schizophrenia and Related Disorders |
US7749694B2 (en) | 2004-12-31 | 2010-07-06 | The Regents Of The University Of California | C-type lectin fold as a scaffold for massive sequence variation |
KR101404512B1 (en) | 2005-01-05 | 2015-01-29 | 에프-스타 비오테크놀로기쉐 포르슝스 운드 엔트비클룽스게스.엠.베.하. | Synthetic immunoglobulin domains with binding properties engineered in regions of the molecule different from the complementarity determining regions |
GB0504767D0 (en) * | 2005-03-08 | 2005-04-13 | Ares Trading Sa | Lipocalin protein |
US7833979B2 (en) * | 2005-04-22 | 2010-11-16 | Amgen Inc. | Toxin peptide therapeutic agents |
US8008453B2 (en) | 2005-08-12 | 2011-08-30 | Amgen Inc. | Modified Fc molecules |
WO2007062676A1 (en) | 2005-12-01 | 2007-06-07 | B.R.A.H.M.S. Aktiengesellschaft | Methods for the diagnosis and treatment of critically ill patients with endothelin, endothelin agonists and adrenomedullin antagonists |
US8445639B2 (en) * | 2006-05-15 | 2013-05-21 | Avidbiotics Corporation | Recombinant bacteriophage and methods for their use |
JP5603070B2 (en) * | 2006-05-15 | 2014-10-08 | アビッドバイオティクス コーポレイション | Modified bacteriocin and method of use thereof |
US7700729B2 (en) * | 2006-05-15 | 2010-04-20 | Avidbiotics Corporation | Modified bacteriocins and methods for their use |
CN101506227A (en) * | 2006-05-26 | 2009-08-12 | 怀卡托灵科有限公司 | OB-fold domains |
HUE030269T2 (en) | 2006-06-26 | 2017-04-28 | Macrogenics Inc | Fc riib-specific antibodies and methods of use thereof |
AT503889B1 (en) | 2006-07-05 | 2011-12-15 | Star Biotechnologische Forschungs Und Entwicklungsges M B H F | MULTIVALENT IMMUNE LOBULINE |
AT503902B1 (en) * | 2006-07-05 | 2008-06-15 | F Star Biotech Forsch & Entw | METHOD FOR MANIPULATING IMMUNE LOBULINS |
WO2008003473A2 (en) * | 2006-07-06 | 2008-01-10 | Merck Patent Gmbh | Compositions and methods for enhancing the efficacy of il-2 mediated immune responses |
EP2423333A1 (en) | 2006-08-25 | 2012-02-29 | Oncotherapy Science, Inc. | Prognostic markers and therapeutic targets for lung cancer |
WO2008048970A2 (en) * | 2006-10-16 | 2008-04-24 | The Arizona Board Of Regents, A Body Corporate Of The State Of Arizona Acting For And On Behalf Of Arizona State University | Synthetic antibodies |
JP5238710B2 (en) | 2006-10-19 | 2013-07-17 | シーエスエル、リミテッド | High affinity antibody antagonist of interleukin-13 receptor alpha 1 |
US8613925B2 (en) | 2006-10-19 | 2013-12-24 | Csl Limited | Anti-IL-13Rα1 antibodies and their uses thereof |
CA2667678A1 (en) * | 2006-10-25 | 2008-07-24 | Amgen Inc. | Toxin peptide therapeutic agents |
NZ598345A (en) | 2006-11-21 | 2013-09-27 | Kalobios Pharmaceuticals Inc | Methods of treating chronic inflammatory diseases using a gm-csf antagonist |
WO2008103378A2 (en) | 2007-02-20 | 2008-08-28 | Merrimack Pharmaceuticals, Inc. | Methods of treating multiple sclerosis by administration of alpha-fetoprotein in combination with an integrin antagonist |
CN101641115B (en) | 2007-03-08 | 2013-04-24 | 卡罗拜奥斯制药公司 | Epha3 antibodies for the treatment of solid tumors |
US11078262B2 (en) | 2007-04-30 | 2021-08-03 | Allergan, Inc. | High viscosity macromolecular compositions for treating ocular conditions |
EP2162540A2 (en) | 2007-05-22 | 2010-03-17 | Amgen Inc. | Compositions and methods for producing bioactive fusion proteins |
EP3241842B1 (en) | 2007-06-26 | 2024-01-31 | F-star Therapeutics Limited | Display of binding agents |
US8470966B2 (en) | 2007-08-10 | 2013-06-25 | Protelica, Inc. | Universal fibronectin type III binding-domain libraries |
US8680019B2 (en) | 2007-08-10 | 2014-03-25 | Protelica, Inc. | Universal fibronectin Type III binding-domain libraries |
US20090176654A1 (en) | 2007-08-10 | 2009-07-09 | Protelix, Inc. | Universal fibronectin type III binding-domain libraries |
AU2008293986A1 (en) * | 2007-08-23 | 2009-03-05 | The Board Of Trustees Of The Leland Stanford Junior University | Modulation of synaptogenesis |
EP2190478B1 (en) | 2007-08-24 | 2016-03-23 | Oncotherapy Science, Inc. | Dkk1 oncogene as therapeutic target for cancer and a diagnosing marker |
RU2010111120A (en) | 2007-08-24 | 2011-09-27 | Онкотерапи Сайенс, Инк. (Jp) | EB13, DLX5, NPTX1 AND CDKN3 AS TARGET TARGETS FOR THERAPY AND DIAGNOSTICS OF LUNG CANCER |
TW200920405A (en) | 2007-08-24 | 2009-05-16 | Oncotherapy Science Inc | PKIB and NAALADL2 for target genes of prostate cancer therapy and diagnosis |
GB2453589A (en) * | 2007-10-12 | 2009-04-15 | King S College London | Protease inhibition |
AR068767A1 (en) | 2007-10-12 | 2009-12-02 | Novartis Ag | ANTIBODIES AGAINST SCLEROSTIN, COMPOSITIONS AND METHODS OF USE OF THESE ANTIBODIES TO TREAT A PATHOLOGICAL DISORDER MEDIATIONED BY SCLEROSTIN |
EA201000718A1 (en) | 2007-11-02 | 2011-06-30 | Новартис Аг | MOLECULES AND METHODS DESIGNED TO MODULATE THE RELATED LIPOPROTEIDES RECEPTOR OF LOW DENSITY PROTEIN 6 (LRP6) |
CL2008003561A1 (en) | 2007-11-30 | 2010-02-05 | Glaxo Group Ltd | Il-13 binding construct comprising at least one antibody single domain (dab) grafted onto a monoclonal antibody (mabdab); polynucleotide and host cell; procedure to prepare the construction; pharmaceutical composition comprising it and used to treat cancer or inflammatory diseases. |
ATE555131T1 (en) | 2008-01-15 | 2012-05-15 | Kalobios Pharmaceuticals Inc | METHOD FOR TREATING BONE DEPOSITIVE DISEASES USING A GM-CSF ANTAGONIST |
AU2009239558B2 (en) | 2008-04-21 | 2013-05-02 | Bio-Rad Laboratories, Inc. | Recombinant deamidated gliadin antigen |
US20120021967A1 (en) * | 2008-04-23 | 2012-01-26 | Arizona Board of Regents, A Body Corporate of the State Of Arizona Acting for and Behalf of Arizona | Synthetic antibodies |
EP2113255A1 (en) | 2008-05-02 | 2009-11-04 | f-star Biotechnologische Forschungs- und Entwicklungsges.m.b.H. | Cytotoxic immunoglobulin |
KR20110010758A (en) | 2008-05-06 | 2011-02-07 | 글락소 그룹 리미티드 | Encapsulation of biologically active agents |
EP3629022A1 (en) | 2008-07-25 | 2020-04-01 | Richard W. Wagner | Protein screening methods |
EP2815766B1 (en) | 2008-08-05 | 2017-07-05 | Novartis AG | Compositions and methods for antibodies targeting complement protein C5 |
WO2010096126A1 (en) | 2008-10-29 | 2010-08-26 | Bg Medicine, Inc. | Galectin-3 immunoassay |
AR074777A1 (en) | 2008-12-19 | 2011-02-09 | Glaxo Group Ltd | PROTEINS OF UNION TO ANTIGEN |
WO2010093814A1 (en) | 2009-02-11 | 2010-08-19 | Kalobios Pharmaceuticals, Inc. | Methods of treating dementia using a gm-csf antagonist |
CA2753287A1 (en) | 2009-02-24 | 2010-09-02 | Glaxo Group Limited | Antigen-binding constructs |
US20110305693A1 (en) | 2009-02-24 | 2011-12-15 | Glaxo Group Limited | Anitigen-binding constructs |
WO2010097394A1 (en) | 2009-02-24 | 2010-09-02 | Glaxo Group Limited | Multivalent and/or multispecific rankl-binding constructs |
AR075715A1 (en) * | 2009-03-05 | 2011-04-20 | Novartis Ag | FORMULATION OF LIOFILIZED ANTIBODY |
WO2010102244A1 (en) | 2009-03-06 | 2010-09-10 | Kalobios Pharmaceuticals, Inc. | Treatment of leukemias and chronic myeloproliferative diseases with antibodies to epha3 |
MA33279B1 (en) | 2009-04-27 | 2012-05-02 | Novartis Ag | COMPOSITIONS AND METHODS FOR INCREASING MUSCLE GROWTH |
EA201101572A1 (en) | 2009-04-27 | 2012-05-30 | Новартис Аг | COMPOSITIONS AND METHODS OF APPLICATION OF THERAPEUTIC ANTIBODIES SPECIFIC TO THE SUB-UNIT OF BETA1 IL-12 RECEPTOR |
WO2010127186A1 (en) | 2009-04-30 | 2010-11-04 | Prognosys Biosciences, Inc. | Nucleic acid constructs and methods of use |
WO2011012646A2 (en) | 2009-07-28 | 2011-02-03 | F. Hoffmann-La Roche Ag | Non-invasive in vivo optical imaging method |
EP2464657B1 (en) | 2009-08-10 | 2015-04-01 | MorphoSys AG | Novel screening strategies for the identification of antibodies or fragments thereof which bind an antigen that has an enzymatic activity |
WO2011029823A1 (en) | 2009-09-09 | 2011-03-17 | Novartis Ag | Monoclonal antibody reactive with cd63 when expressed at the surface of degranulated mast cells |
JP5733784B2 (en) * | 2009-09-24 | 2015-06-10 | 国立大学法人埼玉大学 | Efficient synthesis of cDNA / mRNA-protein conjugates |
US20120282276A1 (en) | 2009-11-05 | 2012-11-08 | The Regents Of The University Of Michigan | Biomarkers predictive of progression of fibrosis |
WO2011080050A2 (en) | 2009-12-11 | 2011-07-07 | Novartis Ag | Binding molecules |
NO2516702T3 (en) | 2009-12-23 | 2018-08-18 | ||
JP5850860B2 (en) | 2010-01-28 | 2016-02-03 | グラクソ グループ リミテッドGlaxo Group Limited | CD127 binding protein |
WO2011098449A1 (en) | 2010-02-10 | 2011-08-18 | Novartis Ag | Methods and compounds for muscle growth |
KR20130010461A (en) | 2010-02-18 | 2013-01-28 | 브리스톨-마이어스 스큅 컴퍼니 | Fibronectin based scaffold domain proteins that bind il-23 |
US20150231215A1 (en) | 2012-06-22 | 2015-08-20 | Randolph J. Noelle | VISTA Antagonist and Methods of Use |
US10745467B2 (en) | 2010-03-26 | 2020-08-18 | The Trustees Of Dartmouth College | VISTA-Ig for treatment of autoimmune, allergic and inflammatory disorders |
CN107098958B (en) | 2010-03-26 | 2021-11-05 | 达特茅斯大学理事会 | VISTA regulatory T cell mediator protein, VISTA binding agent and uses thereof |
US10787701B2 (en) | 2010-04-05 | 2020-09-29 | Prognosys Biosciences, Inc. | Spatially encoded biological assays |
US20190300945A1 (en) | 2010-04-05 | 2019-10-03 | Prognosys Biosciences, Inc. | Spatially Encoded Biological Assays |
PL2556171T3 (en) | 2010-04-05 | 2016-04-29 | Prognosys Biosciences Inc | Spatially encoded biological assays |
KR20130056871A (en) | 2010-04-13 | 2013-05-30 | 브리스톨-마이어스 스큅 컴퍼니 | Fibronectin based scaffold domain proteins that bind pcsk9 |
KR20130066631A (en) | 2010-05-06 | 2013-06-20 | 노파르티스 아게 | Compositions and methods of use for therapeutic low density lipoprotein - related protein 6 (lrp6) multivalent antibodies |
KR20130036012A (en) | 2010-05-07 | 2013-04-09 | 에프. 호프만-라 로슈 아게 | Diagnostic method for the detection of cells ex vivo |
UY33421A (en) | 2010-06-03 | 2011-12-30 | Glaxo Wellcome House | HUMANIZED ANTIGEN UNION PROTEINS |
PT2606070T (en) | 2010-08-20 | 2017-03-31 | Novartis Ag | Antibodies for epidermal growth factor receptor 3 (her3) |
WO2012028683A1 (en) | 2010-09-02 | 2012-03-08 | Novartis Ag | Antibody gel system for sustained drug delivery |
CN103154037A (en) | 2010-10-05 | 2013-06-12 | 诺瓦提斯公司 | Anti-IL 12 Rbeta 1 antibodies and their use in treating autoimmune and inflammatory disorders |
EP2635304B1 (en) | 2010-11-05 | 2017-03-08 | Morphotek, Inc. | Folate receptor alpha as a diagnostic and prognostic marker for folate receptor alpha-expressing cancers |
PT2643352T (en) | 2010-11-23 | 2018-08-01 | Glaxo Group Ltd | Antigen binding proteins to oncostatin m (osm) |
US20130236467A1 (en) | 2010-11-24 | 2013-09-12 | Jeremy Griggs | Multispecific antigen binding proteins targeting hgf |
CN103380143B (en) | 2010-12-22 | 2016-01-06 | 百时美施贵宝公司 | In conjunction with the scaffold domains protein based on fibronectin of IL-23 |
WO2012139110A2 (en) | 2011-04-08 | 2012-10-11 | Prognosys Biosciences, Inc. | Peptide constructs and assay systems |
GB201106254D0 (en) | 2011-04-13 | 2011-05-25 | Frisen Jonas | Method and product |
CN105601741A (en) | 2011-04-15 | 2016-05-25 | 卡姆普根有限公司 | Polypeptides and polynucleotides, and uses thereof for treatment of immune related disorders and cancer |
AR086543A1 (en) | 2011-05-25 | 2014-01-08 | Bg Medicine Inc | GALECTIN-3 INHIBITORS AND METHODS OF USE OF THE SAME, PHARMACEUTICAL COMPOSITION |
ES2953190T3 (en) | 2011-05-27 | 2023-11-08 | Glaxo Group Ltd | BCMA binding proteins (CD269/TNFRSF17) |
WO2012172495A1 (en) | 2011-06-14 | 2012-12-20 | Novartis Ag | Compositions and methods for antibodies targeting tem8 |
CA2840650C (en) | 2011-06-29 | 2019-08-20 | Affinity Biosciences Pty Ltd | Method of protein display |
HUE040276T2 (en) | 2011-07-01 | 2019-02-28 | Novartis Ag | Method for treating metabolic disorders |
US9238689B2 (en) | 2011-07-15 | 2016-01-19 | Morpho Sys AG | Antibodies that are cross-reactive for macrophage migration inhibitory factor (MIF) and D-dopachrome tautomerase (D-DT) |
RU2662671C2 (en) | 2011-11-16 | 2018-07-26 | Адреномед Аг | Antibody to adrenomedullin (adm) or anti-adm antibody element, or anti-adm he-iq framing for application in therapy |
EP2780717B1 (en) | 2011-11-16 | 2016-12-21 | sphingotec GmbH | Adrenomedullin assays and methods for determining mature adrenomedullin |
CA2856136A1 (en) | 2011-11-16 | 2013-05-23 | Adrenomed Ag | Anti-adrenomedullin (adm) antibody or anti-adm antibody fragment or anti-adm non-ig scaffold for reducing the risk of mortality in a patient having a chronic or acute disease or acute condition |
RS58340B1 (en) | 2011-11-16 | 2019-03-29 | Adrenomed Ag | Anti-adrenomedullin (adm) antibody or anti-adm antibody fragment or anti-adm non-ig scaffold for regulating the fluid balance in a patient having a chronic or acute disease |
HUE045940T2 (en) | 2011-11-16 | 2020-01-28 | Adrenomed Ag | Anti-adrenomedullin (adm) antibody or anti-adm antibody fragment or anti-adm non-ig scaffold for use in therapy of an acute disease or acute condition of a patient for stabilizing the circulation |
SG10202006318TA (en) | 2011-11-16 | 2020-08-28 | Adrenomed Ag | Anti-adrenomedullin (adm) antibody or anti-adm antibody fragment or anti-adm non-ig scaffold for prevention or reduction of organ dysfunction or organ failure in a patient having a chronic or acute disease or acute condition |
KR20140103135A (en) | 2011-12-05 | 2014-08-25 | 노파르티스 아게 | Antibodies for epidermal growth factor receptor 3 (her3) directed to domain ii of her3 |
KR102083957B1 (en) | 2011-12-05 | 2020-03-04 | 노파르티스 아게 | Antibodies for epidermal growth factor receptor 3 (her3) |
CN104254777B (en) | 2011-12-05 | 2016-08-24 | 伯乐实验室公司 | Restructuring desamidization gliadin antigen |
GB201121301D0 (en) | 2011-12-12 | 2012-01-25 | Novartis Ag | Method |
AU2012356170B2 (en) | 2011-12-21 | 2016-06-16 | Novartis Ag | Compositions and methods for antibodies targeting Factor P |
WO2013113696A1 (en) | 2012-01-30 | 2013-08-08 | Vib Vzw | Means and method for diagnosis and treatment of alzheimer's disease |
PT2814843T (en) | 2012-02-13 | 2020-06-18 | Agency Science Tech & Res | Il-ãy neutralizing human monoclonal antibodies |
CN103308689B (en) | 2012-03-08 | 2017-04-12 | 思芬构技术有限公司 | A method for predicting the risk of getting cancer or diagnosing cancer in a female subject |
CN103308673B (en) | 2012-03-08 | 2017-05-31 | 思芬构技术有限公司 | For predicting in female subject the method for the risk of cardiovascular event |
CN103308670B (en) | 2012-03-08 | 2017-06-09 | 思芬构技术有限公司 | For predicting the method that object suffers from the risk of diabetes and/or metabolic syndrome |
GB2502127A (en) | 2012-05-17 | 2013-11-20 | Kymab Ltd | Multivalent antibodies and in vivo methods for their production |
US9890215B2 (en) | 2012-06-22 | 2018-02-13 | King's College London | Vista modulators for diagnosis and treatment of cancer |
CA2884704C (en) | 2012-09-07 | 2023-04-04 | Randolph J. Noelle | Vista modulators for diagnosis and treatment of cancer |
JOP20200308A1 (en) | 2012-09-07 | 2017-06-16 | Novartis Ag | IL-18 binding molecules |
TR201809001T4 (en) | 2012-10-02 | 2018-07-23 | Sphingotec Gmbh | A method for diagnosing or monitoring renal function or diagnosing renal dysfunction. |
WO2014072306A1 (en) | 2012-11-08 | 2014-05-15 | F. Hoffmann-La Roche Ag | Her3 antigen binding proteins binding to the beta-hairpin of her3 |
WO2014084859A1 (en) | 2012-11-30 | 2014-06-05 | Novartis Ag | Molecules and methods for modulating tmem16a activities |
PT2928921T (en) | 2012-12-05 | 2021-04-06 | Novartis Ag | Compositions and methods for antibodies targeting epo |
CA2897327A1 (en) | 2013-01-08 | 2014-07-17 | Sphingotec Gmbh | Fasting levels of growth hormone as a predictive marker of cardiovascular risk |
UY35315A (en) | 2013-02-08 | 2014-09-30 | Novartis Ag | ANTI-IL-17A ANTIBODIES AND ITS USE IN THE TREATMENT OF AUTOIMMUNE AND INFLAMMATORY DISORDERS |
AU2014223824B2 (en) | 2013-02-28 | 2020-02-27 | Albert Einstein College Of Medicine, Inc. | Tuberculosis biomarkers and uses thereof |
WO2014141064A1 (en) | 2013-03-13 | 2014-09-18 | Novartis Ag | Notch2 binding molecules for treating respiratory diseases |
WO2014159239A2 (en) | 2013-03-14 | 2014-10-02 | Novartis Ag | Antibodies against notch 3 |
US9005901B2 (en) | 2013-03-15 | 2015-04-14 | Abbott Laboratories | Assay with internal calibration |
CN111233978A (en) | 2013-03-15 | 2020-06-05 | 普罗格诺西斯生物科学公司 | Methods for detecting peptide/MHC/TCR binding |
US9157910B2 (en) | 2013-03-15 | 2015-10-13 | Abbott Laboratories | Assay with increased dynamic range |
EP2970426B1 (en) | 2013-03-15 | 2019-08-28 | Michael C. Milone | Targeting cytotoxic cells with chimeric receptors for adoptive immunotherapy |
US10598674B2 (en) | 2013-03-20 | 2020-03-24 | Sphingotec Gmbh | Adrenomedullin to guide therapy of blood pressure decline |
CN105143250B (en) * | 2013-05-10 | 2020-11-03 | 北京华金瑞清生物医药技术有限公司 | Method for modifying non-antibody protein to generate binding molecule, product thereof and long-acting GLP-1 receptor agonist |
UY35620A (en) | 2013-06-21 | 2015-01-30 | Novartis Ag | ANTIBODIES OF LEXINED OXIDATED LDL RECEIVER 1 AND METHODS OF USE |
AR096601A1 (en) | 2013-06-21 | 2016-01-20 | Novartis Ag | ANTIBODIES OF LEXINED OXIDATED LDL RECEIVER 1 AND METHODS OF USE |
US9868979B2 (en) | 2013-06-25 | 2018-01-16 | Prognosys Biosciences, Inc. | Spatially encoded biological assays using a microfluidic device |
US10208125B2 (en) | 2013-07-15 | 2019-02-19 | University of Pittsburgh—of the Commonwealth System of Higher Education | Anti-mucin 1 binding agents and uses thereof |
BR112016002198A2 (en) | 2013-08-14 | 2017-09-12 | Novartis Ag | Methods of treating sporadic inclusion body myositis |
US10059937B2 (en) | 2013-09-27 | 2018-08-28 | The Board Of Trustees Of The University Of Illinois | Method and kit for generating high affinity binding agents |
US9388222B2 (en) | 2013-10-06 | 2016-07-12 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Modified Pseudomonas exotoxin A |
US10203327B2 (en) | 2013-11-05 | 2019-02-12 | Novartis Ag | Organic compounds |
US10288608B2 (en) | 2013-11-08 | 2019-05-14 | Prognosys Biosciences, Inc. | Polynucleotide conjugates and methods for analyte detection |
US10287354B2 (en) | 2013-12-20 | 2019-05-14 | Novartis Ag | Regulatable chimeric antigen receptor |
MX369173B (en) | 2013-12-24 | 2019-10-30 | Janssen Pharmaceutica Nv | Anti-vista antibodies and fragments. |
US11014987B2 (en) | 2013-12-24 | 2021-05-25 | Janssen Pharmaceutics Nv | Anti-vista antibodies and fragments, uses thereof, and methods of identifying same |
ES2857226T3 (en) | 2014-03-15 | 2021-09-28 | Novartis Ag | Regulable chimeric antigen receptor |
TW201622746A (en) | 2014-04-24 | 2016-07-01 | 諾華公司 | Methods of improving or accelerating physical recovery after surgery for hip fracture |
CN107073109B (en) | 2014-06-11 | 2021-08-06 | 凯西·A·格林 | Use of VISTA agonists and antagonists to inhibit or enhance humoral immunity |
EP3161001A2 (en) | 2014-06-25 | 2017-05-03 | Novartis AG | Antibodies specific for il-17a fused to hyaluronan binding peptide tags |
US10398765B2 (en) | 2014-07-03 | 2019-09-03 | Yale University | Dickkopf2 (Dkk2) inhibition suppresses tumor formation |
US11542488B2 (en) | 2014-07-21 | 2023-01-03 | Novartis Ag | Sortase synthesized chimeric antigen receptors |
EP3174546B1 (en) | 2014-07-31 | 2019-10-30 | Novartis AG | Subset-optimized chimeric antigen receptor-containing t-cells |
WO2016020880A2 (en) | 2014-08-07 | 2016-02-11 | Novartis Ag | Angiopoietin-like 4 antibodies and methods of use |
WO2016020882A2 (en) | 2014-08-07 | 2016-02-11 | Novartis Ag | Angiopoetin-like 4 (angptl4) antibodies and methods of use |
AU2015317608B2 (en) | 2014-09-17 | 2021-03-11 | Novartis Ag | Targeting cytotoxic cells with chimeric receptors for adoptive immunotherapy |
EP3002589A1 (en) | 2014-10-01 | 2016-04-06 | sphingotec GmbH | A method for stratifying a female subject for hormone replacement therapy |
MA41044A (en) | 2014-10-08 | 2017-08-15 | Novartis Ag | COMPOSITIONS AND METHODS OF USE FOR INCREASED IMMUNE RESPONSE AND CANCER TREATMENT |
WO2016059602A2 (en) | 2014-10-16 | 2016-04-21 | Glaxo Group Limited | Methods of treating cancer and related compositions |
HUE049982T2 (en) | 2014-11-14 | 2020-11-30 | Hoffmann La Roche | Antigen binding molecules comprising a tnf family ligand trimer |
EA037590B1 (en) | 2014-11-25 | 2021-04-19 | Бристол-Майерс Сквибб Компани | PD-L1 BINDING POLYPEPTIDES COMPRISING 10Fn3 DOMAIN FOR IMAGING |
BR112017010414A2 (en) | 2014-11-25 | 2018-05-15 | Bristol-Myers Squibb Company | methods and compositions for 18f radiolabeling of biological substances |
WO2016090347A1 (en) | 2014-12-05 | 2016-06-09 | Immunext, Inc. | Identification of vsig8 as the putative vista receptor and its use thereof to produce vista/vsig8 modulators |
UY36449A (en) | 2014-12-19 | 2016-07-29 | Novartis Ag | COMPOSITIONS AND METHODS FOR ANTIBODIES DIRECTED TO BMP6 |
CN116869964A (en) | 2014-12-24 | 2023-10-13 | 耐克西缪恩有限公司 | Nanoparticle compositions and methods for immunotherapy |
SI3280729T1 (en) | 2015-04-08 | 2022-09-30 | Novartis Ag | Cd20 therapies, cd22 therapies, and combination therapies with a cd19 chimeric antigen receptor (car) - expressing cell |
ES2955916T3 (en) | 2015-04-10 | 2023-12-11 | Spatial Transcriptomics Ab | Multiplex analysis of biological specimens of spatially distinguished nucleic acids |
JP6841765B2 (en) | 2015-04-24 | 2021-03-10 | シュピーンゴテック ゲゼルシャフト ミット ベシュレンクテル ハフツング | Methods for Predicting the Risk of Developing Chronic Kidney Disease |
AU2016252008B2 (en) | 2015-04-24 | 2019-06-13 | VIIV Healthcare UK (No.5) Limited | Polypeptides targeting HIV fusion |
EP3549606A1 (en) | 2015-05-28 | 2019-10-09 | Bio-rad Laboratories, Inc. | Affinity ligands and methods relating thereto |
BR112017021484A2 (en) | 2015-06-05 | 2018-07-03 | Novartis Ag | antibodies targeting bone morphogenetic protein 9 (bmp9) and methods for them |
BR112017027870A2 (en) | 2015-06-24 | 2018-08-28 | Janssen Pharmaceutica Nv | antibodies and anti-sight fragments |
JOP20200312A1 (en) | 2015-06-26 | 2017-06-16 | Novartis Ag | Factor xi antibodies and methods of use |
CN108350072B (en) | 2015-08-03 | 2022-05-24 | 诺华股份有限公司 | Methods of treating FGF 21-associated disorders |
EP3347377B1 (en) | 2015-09-09 | 2021-02-17 | Novartis AG | Thymic stromal lymphopoietin (tslp)-binding antibodies and methods of using the antibodies |
AU2016320748B2 (en) | 2015-09-09 | 2019-05-02 | Novartis Ag | Thymic stromal lymphopoietin (TSLP)-binding antibodies and methods of using the antibodies |
US11124791B2 (en) | 2015-09-14 | 2021-09-21 | Arizona Board Of Regents On Behalf Of Arizona State University | Generating recombinant affinity reagents with arrayed targets |
AR106188A1 (en) | 2015-10-01 | 2017-12-20 | Hoffmann La Roche | ANTI-CD19 HUMANIZED HUMAN ANTIBODIES AND METHODS OF USE |
WO2017055404A1 (en) | 2015-10-02 | 2017-04-06 | F. Hoffmann-La Roche Ag | Bispecific antibodies specific for pd1 and tim3 |
EP3356403A2 (en) | 2015-10-02 | 2018-08-08 | H. Hoffnabb-La Roche Ag | Bispecific antibodies specific for a costimulatory tnf receptor |
MX2018004157A (en) | 2015-10-07 | 2019-04-01 | F Hoffmann La Roche Ag | Bispecific antibodies with tetravalency for a costimulatory tnf receptor. |
RU2018119317A (en) | 2015-10-28 | 2019-12-04 | Йейл Юниверсити | HUMANIZED ANTIBODY TO DKK2 AND ITS APPLICATION |
EP3371311B1 (en) | 2015-11-06 | 2021-07-21 | Orionis Biosciences BV | Bi-functional chimeric proteins and uses thereof |
SG10202004259PA (en) | 2015-11-19 | 2020-06-29 | Asclepix Therapeutics Llc | Peptides with anti-angiogenic, anti-lymphangiogenic, and anti-edemic properties and nanoparticle formulations |
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WO2017134305A1 (en) | 2016-02-05 | 2017-08-10 | Orionis Biosciences Nv | Bispecific signaling agents and uses thereof |
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WO2018050902A2 (en) | 2016-09-15 | 2018-03-22 | Quadrucept Bio Limited | Multimers, tetramers & octamers |
AU2017339970A1 (en) | 2016-10-04 | 2019-04-18 | Asclepix Therapeutics, Llc | Compounds and methods for activating Tie2 signaling |
EP3309550A1 (en) | 2016-10-12 | 2018-04-18 | sphingotec GmbH | Method for the detection of apolipoprotein e4 |
CA3040802A1 (en) | 2016-10-24 | 2018-05-03 | Orionis Biosciences Nv | Targeted mutant interferon-gamma and uses thereof |
CN110167962B (en) | 2016-12-16 | 2024-06-07 | 艾德里诺医药公司 | Hyperemic anti-Adrenomedullin (ADM) antibodies or anti-ADM antibody fragments or anti-ADM non-Ig scaffolds for intervention and treatment of patients in need thereof |
US11168147B2 (en) | 2016-12-23 | 2021-11-09 | Novartis Ag | Factor XI antibodies and methods of use |
TW201829469A (en) | 2017-01-03 | 2018-08-16 | 瑞士商赫孚孟拉羅股份公司 | Bispecific antigen binding molecules comprising anti-4-1bb clone 20h4.9 |
JP2020505955A (en) | 2017-02-06 | 2020-02-27 | オリオンズ バイオサイエンス インコーポレイテッド | Targeted modified interferon and uses thereof |
MX2019009255A (en) | 2017-02-06 | 2019-11-05 | Orionis Biosciences Nv | Targeted chimeric proteins and uses thereof. |
WO2018146074A1 (en) | 2017-02-07 | 2018-08-16 | Vib Vzw | Immune-cell targeted bispecific chimeric proteins and uses thereof |
AU2018218557B9 (en) | 2017-02-08 | 2021-06-24 | Novartis Ag | FGF21 mimetic antibodies and uses thereof |
WO2018158398A1 (en) | 2017-03-02 | 2018-09-07 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Antibodies having specificity to nectin-4 and uses thereof |
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CN110573528B (en) | 2017-03-29 | 2023-06-09 | 豪夫迈·罗氏有限公司 | Bispecific antigen binding molecules to costimulatory TNF receptors |
EP3601346A1 (en) | 2017-03-29 | 2020-02-05 | H. Hoffnabb-La Roche Ag | Bispecific antigen binding molecule for a costimulatory tnf receptor |
WO2018178074A1 (en) | 2017-03-29 | 2018-10-04 | F. Hoffmann-La Roche Ag | Trimeric antigen binding molecules specific for a costimulatory tnf receptor |
MA49039A (en) | 2017-04-04 | 2020-02-12 | Hoffmann La Roche | NEW BISPECIFIC ANTIGEN BINDING MOLECULES CAPABLE OF BINDING SPECIFICALLY TO CD40 AND FAP |
SG11201909154SA (en) | 2017-04-05 | 2019-10-30 | Hoffmann La Roche | Bispecific antibodies specifically binding to pd1 and lag3 |
AU2018276361A1 (en) | 2017-05-30 | 2019-12-05 | Sphingotec Gmbh | A method for diagnosing or monitoring kidney function or diagnosing kidney dysfunction |
WO2018229715A1 (en) | 2017-06-16 | 2018-12-20 | Novartis Ag | Compositions comprising anti-cd32b antibodies and methods of use thereof |
AU2018292579A1 (en) | 2017-06-28 | 2019-12-05 | Novartis Ag | Methods for preventing and treating urinary incontinence |
EP3661536A4 (en) | 2017-08-03 | 2021-07-21 | Asclepix Therapeutics, Inc. | Methods for identifying and preparing pharmaceutical agents for activating tie2 receptor |
MX2020003538A (en) | 2017-09-25 | 2020-07-29 | Adrenomed Ag | Anti-adrenomedullin (adm) binder for use in therapy or prevention of symptoms of illness. |
US20220268761A1 (en) | 2017-10-18 | 2022-08-25 | Adrenomed Ag | Therapy monitoring under treatment with an anti-adrenomedullin (adm) binder |
WO2019081504A1 (en) | 2017-10-24 | 2019-05-02 | Sphingotec Gmbh | Selenoprotein p for prediction of a first cardiovascular event |
WO2019081983A1 (en) | 2017-10-25 | 2019-05-02 | Novartis Ag | Antibodies targeting cd32b and methods of use thereof |
CN111542548A (en) | 2017-10-25 | 2020-08-14 | 4Teen4制药有限公司 | DPP3 binding agents directed against and binding to specific DPP3 epitopes and their use in the prevention or treatment of diseases/acute conditions associated with oxidative stress |
EP3703746A1 (en) | 2017-11-01 | 2020-09-09 | F. Hoffmann-La Roche AG | Novel tnf family ligand trimer-containing antigen binding molecules |
US20210324108A1 (en) | 2017-11-01 | 2021-10-21 | Hoffmann-La Roche Inc. | Bispecific 2+1 contorsbodies |
KR20200087236A (en) | 2017-11-22 | 2020-07-20 | 노파르티스 아게 | Reverse binding agents for anti-factor XI/XIa antibodies and uses thereof |
EP3728310A1 (en) | 2017-12-18 | 2020-10-28 | VIIV Healthcare UK (No.5) Limited | Antigen binding polypeptides |
EP3502140A1 (en) | 2017-12-21 | 2019-06-26 | F. Hoffmann-La Roche AG | Combination therapy of tumor targeted icos agonists with t-cell bispecific molecules |
MX2020008208A (en) | 2018-02-05 | 2020-11-09 | Orionis Biosciences Inc | Fibroblast binding agents and use thereof. |
SG11202006686SA (en) | 2018-02-08 | 2020-08-28 | Sphingotec Gmbh | Adrenomedullin (adm) for diagnosis and/or prediction of dementia and anti-adrenomedullin binder for use in therapy or prevention of dementia |
PE20210652A1 (en) | 2018-04-13 | 2021-03-26 | Hoffmann La Roche | HER2 TARGETING ANTIGEN BINDING MOLECULES INCLUDING 4-1BBL |
EP3569614A1 (en) | 2018-05-18 | 2019-11-20 | Julius-Maximilians-Universität Würzburg | Compounds and methods for the immobilization of myostatin-inhibitors on the extracellular matrix by transglutaminase |
TW202015726A (en) | 2018-05-30 | 2020-05-01 | 瑞士商諾華公司 | Entpd2 antibodies, combination therapies, and methods of using the antibodies and combination therapies |
EP3586865A1 (en) | 2018-06-21 | 2020-01-01 | Charité - Universitätsmedizin Berlin | Complement anaphylatoxin binders and their use in treatment of a subject having an ocular wound and/or fibrosis |
CN112424228A (en) | 2018-07-04 | 2021-02-26 | 豪夫迈·罗氏有限公司 | Novel bispecific agonistic 4-1BB antigen-binding molecules |
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US20210347851A1 (en) | 2018-09-28 | 2021-11-11 | Novartis Ag | Cd19 chimeric antigen receptor (car) and cd22 car combination therapies |
CN112654641A (en) | 2018-10-01 | 2021-04-13 | 豪夫迈·罗氏有限公司 | Bispecific antigen binding molecules with trivalent binding to CD40 |
JP7221379B2 (en) | 2018-10-01 | 2023-02-13 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Bispecific antigen-binding molecule comprising anti-FAP clone 212 |
UY38407A (en) | 2018-10-15 | 2020-05-29 | Novartis Ag | TREM2 STABILIZING ANTIBODIES |
WO2020131935A1 (en) | 2018-12-18 | 2020-06-25 | Novartis Ag | Reversal binding agents for anti-factor xi/xia antibodies and uses thereof |
SG11202105176QA (en) | 2018-12-20 | 2021-06-29 | Sphingotec Gmbh | Selenoprotein p in heart failure |
WO2020127618A1 (en) | 2018-12-21 | 2020-06-25 | F. Hoffmann-La Roche Ag | Tumor-targeted agonistic cd28 antigen binding molecules |
TW202030204A (en) | 2018-12-21 | 2020-08-16 | 瑞士商赫孚孟拉羅股份公司 | Tumor-targeted superagonistic cd28 antigen binding molecules |
WO2020128039A2 (en) | 2018-12-21 | 2020-06-25 | 4TEEN4 Pharmaceuticals GmbH | Therapy guidance and/or therapy monitoring for a treatment with angiotensin-receptor-agonist and/or a precursor thereof |
US11254736B2 (en) | 2019-02-15 | 2022-02-22 | Integral Molecular, Inc. | Antibodies comprising a common light chain and uses thereof |
SG11202108281UA (en) | 2019-02-15 | 2021-08-30 | Integral Molecular Inc | Claudin 6 antibodies and uses thereof |
GB201903767D0 (en) | 2019-03-19 | 2019-05-01 | Quadrucept Bio Ltd | Multimers, tetramers & octamers |
WO2020208049A1 (en) | 2019-04-12 | 2020-10-15 | F. Hoffmann-La Roche Ag | Bispecific antigen binding molecules comprising lipocalin muteins |
TW202115124A (en) | 2019-06-26 | 2021-04-16 | 瑞士商赫孚孟拉羅股份公司 | Novel antigen binding molecules binding to cea |
WO2020260326A1 (en) | 2019-06-27 | 2020-12-30 | F. Hoffmann-La Roche Ag | Novel icos antibodies and tumor-targeted antigen binding molecules comprising them |
AU2020328194A1 (en) | 2019-08-15 | 2022-03-17 | Sphingotec Gmbh | A method for diagnosing or monitoring kidney function or diagnosing kidney dysfunction in pediatric patients |
CN114556100A (en) | 2019-08-30 | 2022-05-27 | 4Teen4制药有限公司 | Therapy guidance and/or therapy monitoring for treatment of shock |
TW202124446A (en) | 2019-09-18 | 2021-07-01 | 瑞士商諾華公司 | Combination therapies with entpd2 antibodies |
CN114502590A (en) | 2019-09-18 | 2022-05-13 | 诺华股份有限公司 | ENTPD2 antibodies, combination therapies, and methods of using these antibodies and combination therapies |
EP4072682A1 (en) | 2019-12-09 | 2022-10-19 | Institut National de la Santé et de la Recherche Médicale (INSERM) | Antibodies having specificity to her4 and uses thereof |
EP3871689A1 (en) | 2020-02-26 | 2021-09-01 | sphingotec GmbH | Anti-adm-antibodies binding to the free n-terminus for accelerated transition of adm-gly to bio-adm in patients with adm-gly/ bio-adm ratio above a threshold and combination with vitamin c |
AU2021227279A1 (en) | 2020-02-27 | 2022-10-20 | Adrenomed Ag | Anti-adrenomedullin (ADM) binder for use in therapy of patients in shock |
CA3172349A1 (en) | 2020-02-27 | 2021-09-02 | 4TEEN4 Pharmaceuticals GmbH | Dpp3 for therapy guidance, monitoring and stratification of nt-adm antibodies in patients with shock |
US20230104578A1 (en) | 2020-02-27 | 2023-04-06 | Adrenomed Ag | Anti-adrenomedullin (adm) antibody or anti-adm antibody fragment or anti-adm non-ig scaffold for use in therapy or prevention of shock |
WO2021174045A1 (en) | 2020-02-28 | 2021-09-02 | Bristol-Myers Squibb Company | Radiolabeled fibronectin based scaffolds and antibodies and theranostic uses thereof |
CA3112051A1 (en) | 2020-03-16 | 2021-09-16 | Sphingotec Gmbh | Pro-adrenomedullin or fragment thereof in patients infected with corona virus and treatments with binder against adrenomedullin |
CN115769076A (en) | 2020-03-16 | 2023-03-07 | 4Teen4制药有限公司 | DPP3 in patients infected with coronaviruses |
EP3922993A1 (en) | 2020-06-12 | 2021-12-15 | 4TEEN4 Pharmaceuticals GmbH | Dpp3 in patients infected with coronavirus |
WO2021190980A1 (en) | 2020-03-22 | 2021-09-30 | Quadrucept Bio Limited | Multimers for viral strain evolution |
GB202004514D0 (en) | 2020-03-27 | 2020-05-13 | Inst De Medicina Molecular Joaeo Lobo Antunes | Treatment of Immunosuppressive Cancer |
AR121706A1 (en) | 2020-04-01 | 2022-06-29 | Hoffmann La Roche | OX40 AND FAP-TARGETED BSPECIFIC ANTIGEN-BINDING MOLECULES |
KR20230008751A (en) | 2020-05-12 | 2023-01-16 | 인쎄름 (엥스띠뛰 나씨오날 드 라 쌍떼 에 드 라 흐쉐르슈 메디깔) | Novel methods of treating cutaneous T-cell lymphoma and TFH derived lymphoma |
US20220017637A1 (en) | 2020-06-23 | 2022-01-20 | Hoffmann-La Roche Inc. | Agonistic cd28 antigen binding molecules targeting her2 |
WO2021260064A1 (en) | 2020-06-25 | 2021-12-30 | F. Hoffmann-La Roche Ag | Anti-cd3/anti-cd28 bispecific antigen binding molecules |
JP2023549316A (en) | 2020-11-16 | 2023-11-24 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Combination therapy with CD40 agonists targeting FAP |
EP4023218A1 (en) | 2020-12-02 | 2022-07-06 | S-Form Pharma | Combination therapy for patients having acute and/or persistent dyspnea |
CA3202358A1 (en) | 2020-12-16 | 2022-06-23 | Andreas BOSSHART | Novel slow-release prodrugs |
EP4274658A1 (en) | 2021-01-06 | 2023-11-15 | F. Hoffmann-La Roche AG | Combination therapy employing a pd1-lag3 bispecific antibody and a cd20 t cell bispecific antibody |
WO2022184659A1 (en) | 2021-03-01 | 2022-09-09 | Quadrucept Bio Limited | Antibody domains & multimers |
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AU2022233791A1 (en) | 2021-03-09 | 2023-09-28 | Molecular Partners Ag | Novel darpin based cd123 engagers |
JP2024508969A (en) | 2021-03-09 | 2024-02-28 | モレキュラー パートナーズ アクチェンゲゼルシャフト | Novel DARPin-based CD33 engager |
WO2022243261A1 (en) | 2021-05-19 | 2022-11-24 | F. Hoffmann-La Roche Ag | Agonistic cd40 antigen binding molecules targeting cea |
AR126009A1 (en) | 2021-06-02 | 2023-08-30 | Hoffmann La Roche | CD28 ANTIGEN-BINDING AGONIST MOLECULES THAT TARGET EPCAM |
CN117529664A (en) | 2021-06-18 | 2024-02-06 | 思芬构技术有限公司 | Methods for predicting sepsis and septic shock |
KR20240041912A (en) | 2021-06-29 | 2024-04-01 | 베리솔 게엠베하 | Multiple biomarkers for identifying selenium deficiency in body fluids |
WO2023279092A2 (en) | 2021-07-02 | 2023-01-05 | Genentech, Inc. | Methods and compositions for treating cancer |
CN117715936A (en) | 2021-07-28 | 2024-03-15 | 豪夫迈·罗氏有限公司 | Methods and compositions for treating cancer |
TW202340248A (en) | 2021-12-20 | 2023-10-16 | 瑞士商赫孚孟拉羅股份公司 | Agonistic ltbr antibodies and bispecific antibodies comprising them |
WO2023170296A1 (en) | 2022-03-11 | 2023-09-14 | Inserm (Institut National De La Sante Et De La Recherche Medicale) | Nucleic acid system to specifically reprogram b and t cells and uses thereof |
WO2023175035A1 (en) | 2022-03-15 | 2023-09-21 | Adrenomed Ag | Stable aqueous formulation of an anti-adrenomedullin (adm) antibody or anti-adm antibody fragment |
WO2023186756A1 (en) | 2022-03-28 | 2023-10-05 | F. Hoffmann-La Roche Ag | Interferon gamma variants and antigen binding molecules comprising these |
WO2024008755A1 (en) | 2022-07-04 | 2024-01-11 | Vib Vzw | Blood-cerebrospinal fluid barrier crossing antibodies |
WO2024023368A1 (en) | 2022-07-29 | 2024-02-01 | 4TEEN4 Pharmaceuticals GmbH | Prediction of an increase of dpp3 in a patient with septic shock |
WO2024052503A1 (en) | 2022-09-08 | 2024-03-14 | Institut National de la Santé et de la Recherche Médicale | Antibodies having specificity to ltbp2 and uses thereof |
WO2024056862A1 (en) | 2022-09-15 | 2024-03-21 | Avidicure Ip B.V. | Multispecific antigen binding proteins for tumor-targeting of nk cells and use thereof |
WO2024077118A2 (en) | 2022-10-06 | 2024-04-11 | Bicara Therapeutics Inc. | Multispecific proteins and related methods |
GB202216503D0 (en) | 2022-11-05 | 2022-12-21 | Quadrucept Bio Ltd | Non-human vertebrates & cells |
WO2024126793A1 (en) | 2022-12-15 | 2024-06-20 | 4TEEN4 Pharmaceuticals GmbH | Dpp3 inhibitor for improvement of pulmonary function in critically ill patients |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1998031700A1 (en) * | 1997-01-21 | 1998-07-23 | The General Hospital Corporation | Selection of proteins using rna-protein fusions |
WO1998056915A2 (en) * | 1997-06-12 | 1998-12-17 | Research Corporation Technologies, Inc. | Artificial antibody polypeptides |
WO2000034784A1 (en) * | 1998-12-10 | 2000-06-15 | Phylos, Inc. | Protein scaffolds for antibody mimics and other binding proteins |
Family Cites Families (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5770697A (en) | 1986-11-04 | 1998-06-23 | Protein Polymer Technologies, Inc. | Peptides comprising repetitive units of amino acids and DNA sequences encoding the same |
US5514581A (en) | 1986-11-04 | 1996-05-07 | Protein Polymer Technologies, Inc. | Functional recombinantly prepared synthetic protein polymer |
US6018030A (en) | 1986-11-04 | 2000-01-25 | Protein Polymer Technologies, Inc. | Peptides comprising repetitive units of amino acids and DNA sequences encoding the same |
US5641648A (en) | 1986-11-04 | 1997-06-24 | Protein Polymer Technologies, Inc. | Methods for preparing synthetic repetitive DNA |
US5235041A (en) | 1990-12-28 | 1993-08-10 | Protein Polymer Technologies, Inc. | Purification of structurally ordered recombinant protein polymers |
US5792742A (en) | 1991-06-14 | 1998-08-11 | New York University | Fibrin-binding peptide fragments of fibronectin |
WO1994017097A1 (en) | 1993-01-19 | 1994-08-04 | Regents Of The University Of Minnesota | Synthetic fibronectin fragments as inhibitors of retroviral infections |
GB9618960D0 (en) * | 1996-09-11 | 1996-10-23 | Medical Science Sys Inc | Proteases |
US6159722A (en) * | 1997-12-03 | 2000-12-12 | Boehringer Mannheim Gmbh | Chimeric serine proteases |
IL138668A0 (en) * | 1998-04-03 | 2001-10-31 | Phylos Inc | Addressable protein arrays |
US6818418B1 (en) * | 1998-12-10 | 2004-11-16 | Compound Therapeutics, Inc. | Protein scaffolds for antibody mimics and other binding proteins |
CA2412664A1 (en) * | 2000-06-15 | 2001-12-20 | Board Of Regents The University Of Texas System | Regulatable, catalytically active nucleic acids |
EP2141243A3 (en) * | 2000-10-16 | 2010-01-27 | Brystol-Myers Squibb Company | Protein scaffolds for antibody mimics and other binding proteins |
-
2000
- 2000-02-29 US US09/515,260 patent/US6818418B1/en not_active Expired - Lifetime
-
2001
- 2001-02-28 ES ES01913159T patent/ES2276770T3/en not_active Expired - Lifetime
- 2001-02-28 PT PT01913159T patent/PT1266025E/en unknown
- 2001-02-28 EP EP01913159A patent/EP1266025B1/en not_active Expired - Lifetime
- 2001-02-28 AT AT01913159T patent/ATE346160T1/en active
- 2001-02-28 WO PCT/US2001/006414 patent/WO2001064942A1/en active Search and Examination
- 2001-02-28 AU AU2001241850A patent/AU2001241850B2/en not_active Ceased
- 2001-02-28 JP JP2001563629A patent/JP4829457B2/en not_active Expired - Fee Related
- 2001-02-28 DK DK01913159T patent/DK1266025T3/en active
- 2001-02-28 AU AU4185001A patent/AU4185001A/en active Pending
- 2001-02-28 CA CA002400058A patent/CA2400058A1/en not_active Abandoned
- 2001-02-28 DE DE60124678T patent/DE60124678T2/en not_active Expired - Lifetime
-
2004
- 2004-11-15 US US10/989,723 patent/US20050255548A1/en not_active Abandoned
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1998031700A1 (en) * | 1997-01-21 | 1998-07-23 | The General Hospital Corporation | Selection of proteins using rna-protein fusions |
WO1998056915A2 (en) * | 1997-06-12 | 1998-12-17 | Research Corporation Technologies, Inc. | Artificial antibody polypeptides |
WO2000034784A1 (en) * | 1998-12-10 | 2000-06-15 | Phylos, Inc. | Protein scaffolds for antibody mimics and other binding proteins |
Non-Patent Citations (2)
Title |
---|
KOIDE ET AL.: "The fibronectin type III domain as a scaffold for novel binding proteins", J. MOL. BIOL., vol. 284, 1998, pages 1141 - 1151, XP002944100 * |
NYGREN ET AL.: "Scaffolds for engineering novel binding sites in proteins", ENGINEERING AND DESIGN, vol. 7, 1997, pages 463 - 469, XP002944099 * |
Cited By (233)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7858090B2 (en) | 1997-06-12 | 2010-12-28 | Novartis International Pharmaceutical Ltd. | Artificial antibody polypeptides |
US7981620B2 (en) | 1997-06-12 | 2011-07-19 | Novartis International Pharmaceutical Ltd. | Artificial antibody polypeptides |
US8062858B2 (en) | 1997-06-12 | 2011-11-22 | Novartis International Pharmaceutical Ltd. | Artificial antibody polypeptides |
US9127090B2 (en) | 1997-06-12 | 2015-09-08 | Novartis Ag | Artificial antibody polypeptides |
US8106162B2 (en) | 1997-06-12 | 2012-01-31 | Novartis Ag | Artificial antibody polypeptides |
US7078490B2 (en) | 1997-06-12 | 2006-07-18 | Research Corporation Technologies, Inc. | Artificial antibody polypeptides |
US7119171B2 (en) | 1997-06-12 | 2006-10-10 | Research Corporation Technologies, Inc. | Binding polypeptides |
US7153661B2 (en) | 1997-06-12 | 2006-12-26 | Research Corporation Technologies, Inc. | Artificial antibody polypeptides |
US9605039B2 (en) | 1998-12-10 | 2017-03-28 | Bristol-Myers Squibb Company | Protein scaffolds for antibody mimics and other binding proteins |
US7115396B2 (en) | 1998-12-10 | 2006-10-03 | Compound Therapeutics, Inc. | Protein scaffolds for antibody mimics and other binding proteins |
US8263741B2 (en) | 2000-07-11 | 2012-09-11 | Research Corporation Technologies, Inc. | Artificial antibody polypeptides |
EP1301538B1 (en) | 2000-07-11 | 2015-12-09 | Research Corporation Technologies, Inc | Artificial antibody polypeptides |
EP1356075A4 (en) * | 2000-10-16 | 2005-04-13 | Compound Therapeutics Inc | Protein scaffolds for antibody mimics and other binding proteins |
EP2141243A3 (en) * | 2000-10-16 | 2010-01-27 | Brystol-Myers Squibb Company | Protein scaffolds for antibody mimics and other binding proteins |
EP1356075A2 (en) * | 2000-10-16 | 2003-10-29 | Phylos, Inc. | Protein scaffolds for antibody mimics and other binding proteins |
EP1318195A1 (en) * | 2001-12-10 | 2003-06-11 | CatchMabs B.V. | A structure for presenting desired peptide sequences |
WO2003050283A3 (en) * | 2001-12-10 | 2004-04-01 | Catchmabs B V | A structure for presenting desired peptide sequences |
WO2003050283A2 (en) * | 2001-12-10 | 2003-06-19 | Catchmabs B.V. | A structure for presenting desired peptide sequences |
US8258265B2 (en) | 2002-06-06 | 2012-09-04 | Research Corporation Technologies, Inc. | Reconstituted polypeptides |
US10370440B2 (en) | 2002-06-06 | 2019-08-06 | Research Corporation Technologies, Inc. | Reconstituted polypeptides |
US9051357B2 (en) | 2002-06-06 | 2015-06-09 | Research Corporation Technologies, Inc. | Reconstituted polypeptides |
WO2004092741A3 (en) * | 2003-04-14 | 2005-03-31 | Univ Montana State | Mapping discontinuous antibody or aptamer epitopes for protein structure determination and other applications |
WO2004092741A2 (en) * | 2003-04-14 | 2004-10-28 | Montana State University | Mapping discontinuous antibody or aptamer epitopes for protein structure determination and other applications |
EP2382990A1 (en) | 2003-04-30 | 2011-11-02 | Universität Zürich | Methods for treating cancer using an immunotoxin |
US11168125B2 (en) | 2003-05-06 | 2021-11-09 | Bioverativ Therapeutics Inc. | Immunoglobulin chimeric monomer-dimer hybrids |
US7858739B2 (en) | 2003-12-05 | 2010-12-28 | Bristol-Myers Squibb Company | Inhibitors of type 2 vascular endothelial growth factor receptors |
US8609613B2 (en) | 2003-12-05 | 2013-12-17 | Bristol-Myers Squibb Company | Methods of inhibiting type 2 vascular endothelial growth factor receptors using polypeptides |
US8324362B2 (en) | 2003-12-05 | 2012-12-04 | Bristol-Myers Squibb Company | Nucleic acid encoding 10FN3 polypeptide inhibitors of Type 2 vascular endothelial growth factor receptors |
US7847062B2 (en) | 2003-12-05 | 2010-12-07 | Bristol-Myers Squibb Company | Pharmacokinetic modulation and compositions for modified FN3 polypeptides |
US9328157B2 (en) | 2003-12-05 | 2016-05-03 | Bristol-Myers Squibb Company | Inhibitors of type 2 vascular endothelial growth factor receptors |
US10995131B2 (en) | 2003-12-05 | 2021-05-04 | Bristol-Myers Squibb Company | Libraries of modified fibronectin type III tenth domain-containing polypeptides |
US9862758B2 (en) | 2003-12-05 | 2018-01-09 | Bristol-Myers Quibb Company | Inhibitors of type 2 vascular endothelial growth factor receptors |
US7939265B2 (en) | 2004-01-06 | 2011-05-10 | Badrilla Limited | Agents for and method of quantifying binding |
WO2005066630A3 (en) * | 2004-01-06 | 2005-10-13 | Badrilla Ltd | Agents for and method of quantifying binding |
EP2151453A1 (en) | 2004-06-17 | 2010-02-10 | Amgen Mountain View Inc. | C-met kinase binding proteins |
US8470332B2 (en) | 2006-11-22 | 2013-06-25 | Bristol-Myers Squibb Company | Targeted therapeutics based on engineered proteins for tyrosine kinases receptors, including IGF-IR |
US10221232B2 (en) | 2006-11-22 | 2019-03-05 | Bristol-Myers Squibb Company | Methods of treating cancer by administering IGF-IR binding molecules |
EP3156415A1 (en) | 2006-11-22 | 2017-04-19 | Bristol-Myers Squibb Company | Targeted therapeutics based on engineered proteins for tyrosine kinases receptors, including igf-ir |
EP2727936A1 (en) | 2006-11-22 | 2014-05-07 | Bristol-Myers Squibb Company | Targeted therapeutics based on engineered proteins for tyrosine kinases receptors, including IGF-IR |
US11149077B2 (en) | 2006-11-22 | 2021-10-19 | Bristol-Myers Squibb Company | Targeted therapeutics based on engineered proteins for tyrosine kinases receptors, including IGF-IR |
WO2008097497A2 (en) | 2007-02-02 | 2008-08-14 | Adnexus, A Bristol-Myers Squibb R & D Company | Vegf pathway blockade |
EP2559703A1 (en) | 2007-02-08 | 2013-02-20 | Domantis Limited | Antibody single variable domains against serum albumin |
EP2559704A1 (en) | 2007-02-08 | 2013-02-20 | Domantis Limited | Antibody single variable domains against serum albumin |
EP2559702A1 (en) | 2007-02-08 | 2013-02-20 | Domantis Limited | Antibody single variable domains against serum albumin |
US9938336B2 (en) | 2007-03-12 | 2018-04-10 | Esbatech, An Alcon Biomedical Research Unit Llc | Sequence based engineering and optimization of single chain antibodies |
US8280711B2 (en) | 2007-03-12 | 2012-10-02 | ESBATech, an Alcon Biomedical Research Unit, LLC. | Sequence based engineering and optimization of single chain antibodies |
EP2851091A1 (en) | 2007-04-13 | 2015-03-25 | Dana-Farber Cancer Institute, Inc. | Methods for treating cancer resistant to ERBB therapeutics |
US8715665B2 (en) | 2007-04-13 | 2014-05-06 | The General Hospital Corporation | Methods for treating cancer resistant to ErbB therapeutics |
US11008620B2 (en) | 2007-04-13 | 2021-05-18 | The General Hospital Corporation | Methods for treating cancer resistant to ErbB therapeutics |
US8492328B2 (en) | 2007-05-17 | 2013-07-23 | Bristol-Myers Squibb Company | Biomarkers and methods for determining sensitivity to insulin growth factor-1 receptor modulators |
EP2559771A2 (en) | 2007-05-17 | 2013-02-20 | Bristol-Myers Squibb Company | Biomarkers and methods for determining sensitivity to insulin growth factor-1 receptor modulators |
WO2008144345A2 (en) | 2007-05-17 | 2008-11-27 | Bristol-Myers Squibb Company | Biomarkers and methods for determining sensitivity to insulin growth factor-1 receptor modulators |
US9908945B2 (en) | 2007-06-25 | 2018-03-06 | Esbatech, An Alcon Biomedical Research Unit Llc | Sequence based engineering and optimization of single chain antibodies |
WO2009000099A2 (en) | 2007-06-25 | 2008-12-31 | Esbatech Ag | Methods of modifying antibodies, and modified antibodies with improved functional properties |
US9176129B2 (en) | 2007-10-31 | 2015-11-03 | Medimmune, Llc | Protein scaffolds |
US8633297B2 (en) | 2007-10-31 | 2014-01-21 | Medimmune, Llc | Protein scaffolds |
WO2009062170A1 (en) * | 2007-11-08 | 2009-05-14 | The University Of Chicago | Molecular affinity clamp technology and uses thereof |
US9885050B2 (en) | 2007-11-08 | 2018-02-06 | The University Of Chicago | Molecular affinity clamp technology and uses thereof |
CN101965198A (en) * | 2007-12-27 | 2011-02-02 | 诺瓦提斯公司 | Improved fibronectin-based binding molecules and their use |
WO2009083804A3 (en) * | 2007-12-27 | 2010-04-01 | Novartis Ag | Improved fibronectin-based binding molecules and their use |
WO2009083804A2 (en) * | 2007-12-27 | 2009-07-09 | Novartis Ag | Improved fibronectin-based binding molecules and their use |
US8524244B2 (en) | 2008-02-14 | 2013-09-03 | Bristol-Myers Squibb Company | Targeted therapeutics based on engineered proteins that bind EGFR |
WO2009102421A2 (en) | 2008-02-14 | 2009-08-20 | Bristol-Myers Squibb Company | Targeted therapeutics based on engineered proteins that bind egfr |
US10781247B2 (en) | 2008-02-14 | 2020-09-22 | Bristol-Myers Squibb Company | Targeted therapeutics based on engineered proteins that bind EGFR |
US9920108B2 (en) | 2008-02-14 | 2018-03-20 | Bristol-Myers Squibb Company | Targeted therapeutics based on engineered proteins that bind EGFR |
US9234028B2 (en) | 2008-02-14 | 2016-01-12 | Bristol-Myers Squibb Company | Targeted therapeutics based on engineered proteins that bind EGFR |
US9296810B2 (en) | 2008-05-02 | 2016-03-29 | Novartis Ag | Fibronectin-based binding molecules and uses thereof |
US8221765B2 (en) | 2008-05-22 | 2012-07-17 | Bristol-Myers Squibb Company | Multivalent fibronectin based scaffold domain proteins |
US9902762B2 (en) | 2008-05-22 | 2018-02-27 | Bristol-Myers Squibb Company | Multivalent fibronectin based scaffold domain proteins |
US8728483B2 (en) | 2008-05-22 | 2014-05-20 | Bristol-Myers Squibb Company | Multivalent fibronectin based scaffold domain proteins |
US10774130B2 (en) | 2008-05-22 | 2020-09-15 | Bristol-Myers Squibb Company | Method of treating cancer by administering multivalent fibronectin based scaffold domain proteins |
EP2799448A1 (en) | 2008-05-22 | 2014-11-05 | Bristol-Myers Squibb Company | Multivalent fibronectin based scaffold domain proteins |
US11046757B2 (en) | 2008-06-25 | 2021-06-29 | Novartis Ag | Solubility optimization of immunobinders |
US9556265B2 (en) | 2008-06-25 | 2017-01-31 | Esbatech, An Alcon Biomedical Research Unit Llc | Solubility optimization of immunobinders |
EP3241843A1 (en) | 2008-06-25 | 2017-11-08 | ESBATech, an Alcon Biomedical Research Unit LLC | Solubility optimization of immunobinders |
US10221237B2 (en) | 2008-06-25 | 2019-03-05 | Esbatech, An Alcon Biomedical Research Unit Llc | Solubility optimization of immunobinders |
US9089610B2 (en) | 2008-08-19 | 2015-07-28 | Nektar Therapeutics | Complexes of small-interfering nucleic acids |
US9433684B2 (en) | 2008-08-19 | 2016-09-06 | Nektar Therapeutics | Conjugates of small-interfering nucleic acids |
US10654913B2 (en) | 2008-10-31 | 2020-05-19 | Janssen Biotech, Inc. | Fibronectin type III domain based scaffold compositions, methods and uses |
US10040842B2 (en) | 2008-10-31 | 2018-08-07 | Janssen Biotech, Inc. | Fibronectin type III domain based scaffold compositions, methods and uses |
US11479880B2 (en) | 2008-10-31 | 2022-10-25 | Janssen Biotech, Inc. | Fibronectin type III domain based scaffold compositions, methods and uses |
US8617894B2 (en) | 2008-10-31 | 2013-12-31 | Janssen Biotech, Inc. | Anti-TNF alpha fibronectin type III domain based scaffold compositions, methods and uses |
US8415291B2 (en) | 2008-10-31 | 2013-04-09 | Centocor Ortho Biotech Inc. | Anti-TNF alpha fibronectin type III domain based scaffold compositions, methods and uses |
US9200059B2 (en) | 2008-10-31 | 2015-12-01 | Janssen Biotech, Inc. | Fibronectin type III domain based scaffold compositions, methods and uses |
US8278419B2 (en) | 2008-10-31 | 2012-10-02 | Centocor Ortho Biotech Inc. | Fibronectin type III domain based scaffold compositions, methods and uses |
EP3029064A1 (en) | 2008-11-24 | 2016-06-08 | Bristol-Myers Squibb Company | Bispecific egfr/igfir binding molecules |
US10954286B2 (en) | 2008-11-24 | 2021-03-23 | Bristol-Myers Squibb Company | Bispecific EGFR/IGFIR binding molecules |
US9771411B2 (en) | 2008-11-24 | 2017-09-26 | Bristol-Myers Squibb Company | Method of treating cancer by administering EGFR and EGFR/IGFIR binding molecules |
US10183987B2 (en) | 2008-11-24 | 2019-01-22 | Bristol-Myers Squibb Company | Polynucleotides encoding bispecific EGFR/IGF-IR binding molecules |
WO2010060095A1 (en) | 2008-11-24 | 2010-05-27 | Bristol-Myers Squibb Company | Bispecific egfr/igfir binding molecules |
US8343501B2 (en) | 2008-11-24 | 2013-01-01 | Bristol-Myers Squibb Company | Bispecific EGFR/IGFIR binding molecules |
US9017655B2 (en) | 2008-11-24 | 2015-04-28 | Bristol-Myers Squibb Company | Bispecific EGFR/IGFIR binding molecules |
US10041064B2 (en) | 2008-12-16 | 2018-08-07 | Novartis Ag | Yeast display systems |
US10301616B2 (en) | 2008-12-16 | 2019-05-28 | Novartis Ag | Yeast display systems |
US8067201B2 (en) | 2009-04-17 | 2011-11-29 | Bristol-Myers Squibb Company | Methods for protein refolding |
US9139825B2 (en) | 2009-10-30 | 2015-09-22 | Novartis Ag | Universal fibronectin type III bottom-side binding domain libraries |
WO2011053779A2 (en) | 2009-10-30 | 2011-05-05 | Bristol-Myers Squibb Company | Methods for treating cancer in patients having igf-1r inhibitor resistance |
US10253313B2 (en) | 2009-10-30 | 2019-04-09 | Novartis Ag | Universal fibronectin type III bottom-side binding domain libraries |
EP2679234A2 (en) | 2009-12-02 | 2014-01-01 | Amgen Inc. | Binding proteins that bind to human FGFR1C, human beta-klotho and both human FGFR1C and human beta-klotho |
WO2011068893A1 (en) | 2009-12-02 | 2011-06-09 | Amgen Inc. | BINDING PROTEINS THAT BIND TO HUMAN FGFR1C, HUMAN β-KLOTHO AND BOTH HUMAN FGFR1C AND HUMANβ-KLOTHO |
US9212231B2 (en) | 2010-04-13 | 2015-12-15 | Medimmune, Llc | TRAIL R2-specific multimeric scaffolds |
US8569227B2 (en) | 2010-04-30 | 2013-10-29 | Janssen Biotech, Inc. | Stabilized fibronectin domain compositions, methods and uses |
US9982253B2 (en) | 2010-04-30 | 2018-05-29 | Janssen Biotech, Inc. | Stabilized fibronectin domain compositions, methods and uses |
US9234029B2 (en) | 2010-04-30 | 2016-01-12 | Janssen Biotech, Inc. | Stabilized fibronectin domain compositions, methods and uses |
WO2011140086A2 (en) | 2010-05-03 | 2011-11-10 | Bristol-Myers Squibb Company | Serum albumin binding molecules |
US8969289B2 (en) | 2010-05-03 | 2015-03-03 | Bristol-Myers Squibb Company | Serum albumin binding molecules |
US9540424B2 (en) | 2010-05-03 | 2017-01-10 | Bristol-Myers Squibb Company | Serum albumin binding molecules |
US10934572B2 (en) | 2010-05-03 | 2021-03-02 | Bristol-Myers Squibb Company | Serum albumin binding molecules |
US10221438B2 (en) | 2010-05-03 | 2019-03-05 | Bristol-Myers Squibb Company | Serum albumin binding molecules |
US9428583B2 (en) | 2010-05-06 | 2016-08-30 | Novartis Ag | Compositions and methods of use for therapeutic low density lipoprotein-related protein 6 (LRP6) multivalent antibodies |
US9562089B2 (en) | 2010-05-26 | 2017-02-07 | Bristol-Myers Squibb Company | Fibronectin based scaffold proteins having improved stability |
US10273286B2 (en) | 2010-05-26 | 2019-04-30 | Bristol-Myers Squibb Company | Fibronectin based scaffold proteins having improved stability |
EP3091028A1 (en) | 2010-05-26 | 2016-11-09 | Bristol-Myers Squibb Company | Fibronectin based scaffold proteins having improved stability |
WO2011150133A2 (en) | 2010-05-26 | 2011-12-01 | Bristol-Myers Squibb Company | Fibronectin based scaffold proteins having improved stability |
US11161893B2 (en) | 2010-05-26 | 2021-11-02 | Bristol-Myers Squibb Company | Fibronectin based scaffold proteins having improved stability |
US9856468B2 (en) | 2010-07-09 | 2018-01-02 | Bioverativ Therapeutics Inc. | Processable single chain molecules and polypeptides made using same |
EP3560962A1 (en) | 2010-07-09 | 2019-10-30 | Bioverativ Therapeutics Inc. | Processable single chain molecules and polypeptides made using same |
US10927362B2 (en) | 2010-07-09 | 2021-02-23 | Bioverativ Therapeutics Inc. | Processable single chain molecules and polypeptides made using same |
US10968442B2 (en) | 2010-07-09 | 2021-04-06 | Bioverativ Therapeutics Inc. | Chimeric clotting factors |
WO2012006635A1 (en) | 2010-07-09 | 2012-01-12 | Biogen Idec Hemophilia Inc. | Processable single chain molecules and polypeptides made using same |
US9469676B2 (en) | 2011-04-13 | 2016-10-18 | Bristol-Myers Squibb Company | Fc fusion proteins comprising novel linkers or arrangements |
US10214579B2 (en) | 2011-04-13 | 2019-02-26 | Bristol-Myers Squibb Company | Fc fusion proteins comprising novel linkers or arrangements |
EP3144320A1 (en) | 2011-04-13 | 2017-03-22 | Bristol-Myers Squibb Company | Fc fusion proteins comprising novel linkers or arrangements |
EP3415528A2 (en) | 2011-04-13 | 2018-12-19 | Bristol-Myers Squibb Company | Fc fusion proteins comprising novel linkers or arrangements |
EP3896083A1 (en) | 2011-04-13 | 2021-10-20 | Bristol-Myers Squibb Company | Fc fusion proteins comprising novel linkers or arrangements |
WO2012142515A2 (en) | 2011-04-13 | 2012-10-18 | Bristol-Myers Squibb Company | Fc fusion proteins comprising novel linkers or arrangements |
US10150962B2 (en) | 2011-05-17 | 2018-12-11 | Bristol-Myers Squibb Company | Methods for the selection of binding proteins |
EP3318880A1 (en) | 2011-05-17 | 2018-05-09 | Bristol-Myers Squibb Company | Improved methods for the selection of binding proteins |
US11913137B2 (en) | 2011-05-17 | 2024-02-27 | Bristol-Myers Squibb Company | Methods for the selection of binding proteins |
US9347058B2 (en) | 2011-05-17 | 2016-05-24 | Bristol-Myers Squibb Company | Methods for the selection of binding proteins |
WO2012158678A1 (en) | 2011-05-17 | 2012-11-22 | Bristol-Myers Squibb Company | Methods for maintaining pegylation of polypeptides |
WO2012158739A1 (en) | 2011-05-17 | 2012-11-22 | Bristol-Myers Squibb Company | Improved methods for the selection of binding proteins |
US10898538B2 (en) | 2011-05-17 | 2021-01-26 | Bristol-Myers Squibb Company | Methods for maintaining pegylation of polypeptides |
US11060085B2 (en) | 2011-05-17 | 2021-07-13 | Bristol-Myers Squibb Company | Methods for the selection of binding proteins |
EP3939613A1 (en) | 2011-08-11 | 2022-01-19 | ONO Pharmaceutical Co., Ltd. | Therapeutic agent for autoimmune diseases comprising pd-1 agonist |
WO2013022091A1 (en) | 2011-08-11 | 2013-02-14 | 小野薬品工業株式会社 | Therapeutic agent for autoimmune diseases comprising pd-1 agonist |
US9897612B2 (en) | 2011-09-27 | 2018-02-20 | Janssen Biotech, Inc. | Fibronectin type III repeat based protein scaffolds with alternative binding surfaces |
US9200273B2 (en) | 2011-09-27 | 2015-12-01 | Janssen Biotech, Inc. | Fibronectin type III repeat based protein scaffolds with alternative binding surfaces |
US10571472B2 (en) | 2011-09-27 | 2020-02-25 | Janssen Biotech, Inc. | Fibronectin type III repeat based protein scaffolds with alternative binding surfaces |
US11555062B2 (en) | 2011-10-11 | 2023-01-17 | Viela Bio, Inc. | Methods of administering a composition comprising a Tn3 scaffold and a CD40L-specific monomer |
WO2013056069A1 (en) | 2011-10-13 | 2013-04-18 | Bristol-Myers Squibb Company | Methods for selecting and treating cancer in patients with igf-1r/ir inhibitors |
US9416170B2 (en) | 2011-10-31 | 2016-08-16 | Bristol-Myers Squibb Company | Fibronectin binding domains with reduced immunogenicity |
US11279751B2 (en) | 2011-10-31 | 2022-03-22 | Bristol-Myers Squibb Company | Fibronectin binding domains with reduced immunogenicity |
US11408093B2 (en) | 2011-10-31 | 2022-08-09 | Bristol-Myers Squibb Company | Fibronectin binding domains with reduced immunogenicity |
WO2013067029A2 (en) | 2011-10-31 | 2013-05-10 | Bristol-Myers Squibb Company | Fibronectin binding domains with reduced immunogenicity |
US9765132B2 (en) | 2011-10-31 | 2017-09-19 | Bristol-Myers Squibb Company | Fibronectin binding domains with reduced immunogenicity |
US10604556B2 (en) | 2011-10-31 | 2020-03-31 | Bristol-Myers Squibb Company | Fibronectin binding domains with reduced immunogenicity |
US9522951B2 (en) | 2011-10-31 | 2016-12-20 | Bristol-Myers Squibb Company | Fibronectin binding domains with reduced immunogenicity |
US10464995B2 (en) | 2011-10-31 | 2019-11-05 | Bristol-Myers Squibb Company | Fibronectin binding domains with reduced immunogenicity |
USRE47860E1 (en) | 2011-11-04 | 2020-02-18 | Novartis Ag | Methods of treating cancer with low density lipoprotein-related protein 6 (LRP6)—half life extender constructs |
US9173960B2 (en) | 2011-11-04 | 2015-11-03 | Novartis Ag | Methods of treating cancer with low density lipoprotein-related protein 6 (LRP6)—half life extender constructs |
WO2013106577A2 (en) | 2012-01-10 | 2013-07-18 | Biogen Idec Ma Inc. | Enhancement of transport of therapeutic molecules across the blood brain barrier |
US11827701B2 (en) | 2012-05-23 | 2023-11-28 | argenx BV | IL-6 binding molecules |
US11117959B2 (en) | 2012-05-23 | 2021-09-14 | Argenx Bvba | IL-6 binding molecules |
WO2013175276A1 (en) | 2012-05-23 | 2013-11-28 | Argen-X B.V | Il-6 binding molecules |
WO2013175427A1 (en) | 2012-05-23 | 2013-11-28 | Argen-X B.V. | Il-6 binding molecules |
US10183995B2 (en) | 2012-05-23 | 2019-01-22 | Argen-X N.V. | IL-6 binding molecules |
EP3835310A1 (en) * | 2012-09-13 | 2021-06-16 | Bristol-Myers Squibb Company | Fibronectin based scaffold domain proteins that bind to myostatin |
US8853154B2 (en) | 2012-09-13 | 2014-10-07 | Bristol-Myers Squibb Company | Fibronectin based scaffold domain proteins that bind to myostatin |
EP3564258A1 (en) * | 2012-09-13 | 2019-11-06 | Bristol-Myers Squibb Company | Fibronectin based scaffold domain proteins that bind to myostatin |
WO2014043344A1 (en) * | 2012-09-13 | 2014-03-20 | Bristol-Myers Squibb Company | Fibronectin based scaffold domain proteins that bind to myostatin |
US9662373B2 (en) | 2012-09-13 | 2017-05-30 | Bristol-Myers Squibb Company | Fibronectin based scaffold domain proteins that bind to myostatin |
US8993265B2 (en) | 2012-09-13 | 2015-03-31 | Bristol-Myers Squibb Company | Fibronectin based scaffold domain proteins that bind to myostatin |
US8933199B2 (en) | 2012-09-13 | 2015-01-13 | Bristol-Myers Squibb Company | Fibronectin based scaffold domain proteins that bind to myostatin |
TWI633117B (en) * | 2012-09-13 | 2018-08-21 | 必治妥美雅史谷比公司 | Fibronectin based scaffold domain proteins that bind to myostatin |
EA033536B1 (en) * | 2012-09-13 | 2019-10-31 | Bristol Myers Squibb Co | Fibronectin based scaffold domain proteins that bind to myostatin |
AU2018201273B2 (en) * | 2012-09-13 | 2020-04-09 | Bristol-Myers Squibb Company | Fibronectin based scaffold domain proteins that bind to myostatin |
US11813315B2 (en) | 2012-09-13 | 2023-11-14 | Bristol-Myers Squibb Company | Fibronectin based scaffold domain proteins that bind to myostatin |
US10406212B2 (en) | 2012-09-13 | 2019-09-10 | Bristol-Myers Squibb Company | Fibronectin based scaffold domain proteins that bind to myostatin |
US9493546B2 (en) | 2012-09-13 | 2016-11-15 | Bristol-Myers Squibb Company | Fibronectin based scaffold domain proteins that bind to myostatin |
TWI694085B (en) | 2012-09-13 | 2020-05-21 | 美商必治妥美雅史谷比公司 | Fibronectin based scaffold domain proteins that bind to myostatin |
IL270110B (en) * | 2012-09-13 | 2022-08-01 | Bristol Myers Squibb Co | Fibronectin based scaffold domain proteins that bind to myostatin |
US10245302B2 (en) | 2012-09-13 | 2019-04-02 | Bristol-Myers Squibb Company | Fibronectin based scaffold domain proteins that bind to myostatin |
WO2014120891A2 (en) | 2013-02-01 | 2014-08-07 | Bristol-Myers Squibb Company | Fibronectin based scaffold proteins |
EP2951206A2 (en) * | 2013-02-01 | 2015-12-09 | Bristol-Myers Squibb Company | Fibronectin based scaffold proteins |
US11447538B2 (en) | 2013-02-01 | 2022-09-20 | Bristol-Myers Squibb Company | Fibronectin based scaffold proteins |
US10787498B2 (en) | 2013-02-06 | 2020-09-29 | Bristol-Myers Squibb Company | Fibronectin type III domain proteins with enhanced solubility |
US11512124B2 (en) | 2013-02-06 | 2022-11-29 | Bristol-Myers Squibb Company | Fibronectin type III domain proteins with enhanced solubility |
US11345722B2 (en) | 2013-02-12 | 2022-05-31 | Bristol-Myers Squibb Company | High pH protein refolding methods |
US10065987B2 (en) | 2013-02-12 | 2018-09-04 | Bristol-Myers Squibb Company | High pH protein refolding methods |
WO2014165093A2 (en) | 2013-03-13 | 2014-10-09 | Bristol-Myers Squibb Company | Fibronectin based scaffold domains linked to serum albumin or a moiety binding thereto |
WO2015143156A1 (en) | 2014-03-20 | 2015-09-24 | Bristol-Myers Squibb Company | Stabilized fibronectin based scaffold molecules |
US11203630B2 (en) | 2014-03-20 | 2021-12-21 | Bristol-Myers Squibb Company | Serum albumin-binding fibronectin type III domains |
EP3647322A1 (en) | 2014-03-20 | 2020-05-06 | Bristol-Myers Squibb Company | Stabilized fibronectin based scaffold molecules |
US11407815B2 (en) | 2014-03-20 | 2022-08-09 | Bristol-Myers Squibb Company | Stabilized fibronectin based scaffold molecules |
US10442851B2 (en) | 2014-03-20 | 2019-10-15 | Bristol-Myers Squibb Company | Serum albumin-binding fibronectin type III domains |
US10450363B2 (en) | 2014-03-20 | 2019-10-22 | Bristol-Myers Squibb Company | Stabilized fibronectin based scaffold molecules |
US10766946B2 (en) | 2015-09-23 | 2020-09-08 | Bristol-Myers Squibb Company | Fast-off rate serum albumin binding fibronectin type III domains |
EP3733698A1 (en) | 2015-09-23 | 2020-11-04 | Bristol-Myers Squibb Company | Glypican-3 binding fibronectin based scafflold molecules |
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WO2017165464A1 (en) | 2016-03-21 | 2017-09-28 | Elstar Therapeutics, Inc. | Multispecific and multifunctional molecules and uses thereof |
US11291721B2 (en) | 2016-03-21 | 2022-04-05 | Marengo Therapeutics, Inc. | Multispecific and multifunctional molecules and uses thereof |
WO2017178572A1 (en) | 2016-04-13 | 2017-10-19 | Vivia Biotech, S.L | Ex vivo bite-activated t cells |
WO2017210598A1 (en) | 2016-06-03 | 2017-12-07 | Amgen Inc. | Compositions and methods for treating an articular disorder |
US11673971B2 (en) | 2016-09-23 | 2023-06-13 | Marengo Therapeutics, Inc. | Multispecific antibody molecules comprising lambda and kappa light chains |
WO2018057955A1 (en) | 2016-09-23 | 2018-03-29 | Elstar Therapeutics, Inc. | Multispecific antibody molecules comprising lambda and kappa light chains |
US11447539B2 (en) | 2016-12-14 | 2022-09-20 | Janssen Biotech, Inc. | PD-L1 binding fibronectin type III domains |
US11345739B2 (en) | 2016-12-14 | 2022-05-31 | Janssen Biotech, Inc | CD137 binding fibronectin type III domains |
US11932680B2 (en) | 2016-12-14 | 2024-03-19 | Janssen Biotech, Inc. | CD8A-binding fibronectin type III domains |
US11299534B2 (en) | 2016-12-14 | 2022-04-12 | Janssen Biotech, Inc. | CD8A-binding fibronectin type III domains |
WO2018151820A1 (en) | 2017-02-16 | 2018-08-23 | Elstar Therapeutics, Inc. | Multifunctional molecules comprising a trimeric ligand and uses thereof |
WO2018195283A1 (en) | 2017-04-19 | 2018-10-25 | Elstar Therapeutics, Inc. | Multispecific molecules and uses thereof |
EP4328241A2 (en) | 2017-04-28 | 2024-02-28 | Marengo Therapeutics, Inc. | Multispecific molecules comprising a non-immunoglobulin heterodimerization domain and uses thereof |
WO2018201047A1 (en) | 2017-04-28 | 2018-11-01 | Elstar Therapeutics, Inc. | Multispecific molecules comprising a non-immunoglobulin heterodimerization domain and uses thereof |
WO2018222901A1 (en) | 2017-05-31 | 2018-12-06 | Elstar Therapeutics, Inc. | Multispecific molecules that bind to myeloproliferative leukemia (mpl) protein and uses thereof |
CN111065407A (en) * | 2017-06-16 | 2020-04-24 | 普罗特利卡公司 | Fibronectin binding domain chimeric antigen receptors and methods of use thereof |
WO2019035938A1 (en) | 2017-08-16 | 2019-02-21 | Elstar Therapeutics, Inc. | Multispecific molecules that bind to bcma and uses thereof |
WO2019113464A1 (en) | 2017-12-08 | 2019-06-13 | Elstar Therapeutics, Inc. | Multispecific molecules and uses thereof |
US11680091B2 (en) | 2018-02-23 | 2023-06-20 | The University Of Chicago | Methods and composition involving thermophilic fibronectin type III (FN3) monobodies |
WO2019178362A1 (en) | 2018-03-14 | 2019-09-19 | Elstar Therapeutics, Inc. | Multifunctional molecules that bind to calreticulin and uses thereof |
WO2019178364A2 (en) | 2018-03-14 | 2019-09-19 | Elstar Therapeutics, Inc. | Multifunctional molecules and uses thereof |
US11845797B2 (en) | 2018-07-03 | 2023-12-19 | Marengo Therapeutics, Inc. | Anti-TCR antibody molecules and uses thereof |
DE202019005887U1 (en) | 2018-07-03 | 2023-06-14 | Marengo Therapeutics, Inc. | Anti-TCR antibody molecules and uses thereof |
US11965025B2 (en) | 2018-07-03 | 2024-04-23 | Marengo Therapeutics, Inc. | Method of treating solid cancers with bispecific interleukin-anti-TCRß molecules |
WO2020010250A2 (en) | 2018-07-03 | 2020-01-09 | Elstar Therapeutics, Inc. | Anti-tcr antibody molecules and uses thereof |
US20210396761A1 (en) * | 2018-07-20 | 2021-12-23 | Aicuris Gmbh & Co. Kg | Methods for screening and identifying agents that inhibit or modulate the nuclear egress complex of herpesviruses |
WO2020016433A1 (en) * | 2018-07-20 | 2020-01-23 | Aicuris Gmbh & Co. Kg | Methods for screening and identifying agents that inhibit or modulate the nuclear egress complex of herpesviruses |
WO2020069372A1 (en) | 2018-09-27 | 2020-04-02 | Elstar Therapeutics, Inc. | Csf1r/ccr2 multispecific antibodies |
WO2020172605A1 (en) | 2019-02-21 | 2020-08-27 | Elstar Therapeutics, Inc. | Antibody molecules that bind to nkp30 and uses thereof |
WO2020172598A1 (en) | 2019-02-21 | 2020-08-27 | Elstar Therapeutics, Inc. | Multifunctional molecules that bind to t cells and uses thereof to treat autoimmune disorders |
WO2020172601A1 (en) | 2019-02-21 | 2020-08-27 | Elstar Therapeutics, Inc. | Multifunctional molecules that bind to calreticulin and uses thereof |
WO2020172571A1 (en) | 2019-02-21 | 2020-08-27 | Elstar Therapeutics, Inc. | Multifunctional molecules that bind to t cell related cancer cells and uses thereof |
WO2020172596A1 (en) | 2019-02-21 | 2020-08-27 | Elstar Therapeutics, Inc. | Anti-tcr antibody molecules and thereof |
WO2021006199A1 (en) | 2019-07-05 | 2021-01-14 | 小野薬品工業株式会社 | Treatment of hematologic cancer with pd-1/cd3 dual specificity protein |
WO2021025140A1 (en) | 2019-08-08 | 2021-02-11 | 小野薬品工業株式会社 | Dual-specific protein |
US11781138B2 (en) | 2019-10-14 | 2023-10-10 | Aro Biotherapeutics Company | FN3 domain-siRNA conjugates and uses thereof |
US11628222B2 (en) | 2019-10-14 | 2023-04-18 | Aro Biotherapeutics Company | CD71 binding fibronectin type III domains |
WO2021138407A2 (en) | 2020-01-03 | 2021-07-08 | Marengo Therapeutics, Inc. | Multifunctional molecules that bind to cd33 and uses thereof |
WO2021217085A1 (en) | 2020-04-24 | 2021-10-28 | Marengo Therapeutics, Inc. | Multifunctional molecules that bind to t cell related cancer cells and uses thereof |
WO2022046920A2 (en) | 2020-08-26 | 2022-03-03 | Marengo Therapeutics, Inc. | Multifunctional molecules that bind to calreticulin and uses thereof |
WO2022046922A2 (en) | 2020-08-26 | 2022-03-03 | Marengo Therapeutics, Inc. | Antibody molecules that bind to nkp30 and uses thereof |
WO2022047046A1 (en) | 2020-08-26 | 2022-03-03 | Marengo Therapeutics, Inc. | Methods of detecting trbc1 or trbc2 |
WO2022216993A2 (en) | 2021-04-08 | 2022-10-13 | Marengo Therapeutics, Inc. | Multifuntional molecules binding to tcr and uses thereof |
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CA2400058A1 (en) | 2001-09-07 |
US6818418B1 (en) | 2004-11-16 |
PT1266025E (en) | 2007-02-28 |
EP1266025A1 (en) | 2002-12-18 |
JP2003525487A (en) | 2003-08-26 |
AU4185001A (en) | 2001-09-12 |
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AU2001241850B2 (en) | 2006-09-07 |
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US20050255548A1 (en) | 2005-11-17 |
ATE346160T1 (en) | 2006-12-15 |
EP1266025B1 (en) | 2006-11-22 |
DE60124678D1 (en) | 2007-01-04 |
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