WO2001000237A1 - Synthetic peptide immunogens and antibodies thereto - Google Patents
Synthetic peptide immunogens and antibodies thereto Download PDFInfo
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- WO2001000237A1 WO2001000237A1 PCT/US2000/017609 US0017609W WO0100237A1 WO 2001000237 A1 WO2001000237 A1 WO 2001000237A1 US 0017609 W US0017609 W US 0017609W WO 0100237 A1 WO0100237 A1 WO 0100237A1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- 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
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/70596—Molecules with a "CD"-designation not provided for elsewhere
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- 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
- C07K14/745—Blood coagulation or fibrinolysis factors
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- 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
- C07K16/36—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against blood coagulation factors
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/40—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against enzymes
Definitions
- the present invention generally relates to methods for selecting peptide immunogens to produce antibodies to a target protein which exhibits amino acid variation between species. More specifically, the present invention relates to methods of selecting peptide immunogens and producing species-specific antibodies to a mammalian tissue factor, and antibodies produced by those methods.
- Antibodies to protein antigens have been used for many years for numerous research and diagnostic purposes. Antibodies can be made to specific regions of a protein by injecting a small peptide portion of the protein (5-50 amino acids) which has been conjugated to an immunogenic carrier (such as keyhole limpet hemocyanin [KLH] ) . The immunized animal will recognize the peptide as an epitope of the immunogenic carrier.
- an immunogenic carrier such as keyhole limpet hemocyanin [KLH]
- KLH keyhole limpet hemocyanin
- This approach has advantages over the more common methods involving the injection of the whole protein. For example, this approach allows antibody reagents to be made which bind to specific, predictable domains of the protein of interest.
- These anti- peptide antibodies are useful for epitope mapping in structure/function studies, immunohistochemical localization, and immunoaffinity purification of the protein of interest.
- One particular protein of interest for this invention is mammalian tissue factor.
- Tissue factor is a transmembrane glycoprotein which plays a critical role in coagulation (blood clotting) .
- Normal coagulation begins when vascular endothelium is damaged. Damaged vascular endothelium exposes TF to plasma, allowing TF to bind to factor VII, a serine protease zy ogen.
- factor VII a serine protease zy ogen.
- the binding of TF to factor VII, and subsequent activation to factor Vila is the primary step for coagulation initiation. Subsequent conformational changes and Ca2+ bridge formation allow the
- TF/FVIIa complex to bind to and to activate factor X.
- Activated factor X converts prothrombin into its active serine protease, thrombin.
- Thrombin converts fibrinogen into fibrin.
- fibrin fibers associate with one another to form a clot, or a platelet-fibrin network that prevents further bleeding.
- the amino acid sequence of TF protein between mice, rabbits, and humans is highly conserved. Pawashe et al . , 1991, Thromb . Haemost . 66 . : 315.
- the crystal structure of TF consists of two immunoglobulin-like domains with an extensive interdomain interface region which contains the binding site for factor VII. Harlos et al . , 1994, Nature 370:662.
- TF is expressed in various tissues such as cardiac myocytes, renal glomeruli, the granular layer of the epidermis, the epithelium of oropharynx and vagina, and intestinal, urinary bladder, respiratory mucosa, and at tissue barriers between the body and the environment.
- Tissue factor has also been implicated in the pathogenesis of various infectious, cardiovascular and neoplastic diseases. Excessive coagulation, for example, leads to stroke and ischemia, resulting in infarction of tissues.
- Acute myeloblastic leukemias are known to express TF, resulting in disseminated intravascular coagulation (DIC) , a disorder characterized by excessive bleeding due to rapid consumption of platelets and disruption of fibrin polymerization. Id .
- DIC disseminated intravascular coagulation
- Anti -peptide antibodies to TF has been made.
- polyclonal antibodies were made to human TF regions which allow them to inhibit TF binding to factor VII.
- anti-peptide monoclonal antibodies were made to the C-terminal cytoplasmic domain of human TF . Carson et al . , 1992, Blood Coagulation and Fibrinolysis 3 : 119 .
- Antibodies made to proteins such as TF which have extensive homology between species would not be expected to exhibit species-specificity. This has been shown to be the case with TF, where monoclonal antibodies to human tissue factor cross-react and inhibit baboon TF . Taylor et al . , 1991, Circulation Shock 33_:127. This could be a problem, e.g., where the antibodies are used to diagnose diseases and are required to distinguish between two pathogenic species. Proteins such as TF with extensive sequence homology among vertebrates are also limited in the range of antibodies they would be expected to induce.
- the immune system of the immunizing animal would recognize the homologous regions of the injected protein as a "self" epitope and would thus only produce antibodies to immunodominant regions of the injected protein which are not homologous with the injected animal's analogous protein.
- murine monoclonal antibodies to whole human TF could only be produced to regions of the human TF which are dissimilar to mouse TF . If antibodies are desired to many regions of the TF protein, for example for structure-function studies, the results would likely be disappointing. Therefore, there is a need for improved methods for making antibodies to proteins which show extensive amino acid homology between species, where the antibodies need to be species-specific or where antibodies are desired to several regions of vertebrate proteins .
- a more specific object of the invention is the provision of methods for determining regions of tissue factor for use as immunogens to produce antibodies which are species-specific.
- Another object of the invention is the provision of antibodies to tissue factor made using the above methods .
- the present invention is directed to a method for selecting a peptide immunogen of a target protein from a first species, wherein the target protein comprises an amino acid sequence which varies between the first species and a second species.
- the method comprises (a) identifying peptide regions of the amino acid sequence of the target protein from the first species which has a hydrophilicity value greater than 0, wherein the regions are 5-50 amino acids in length, and (b) selecting a peptide immunogen from regions identified in step (a) which have at least 1 nonhomologous amino acid between the first species and the second species .
- the present invention is also directed to a method of making an antibody which is specific for a target protein of a first species, wherein the target protein comprises an amino acid sequence which varies between the first species and the second species.
- the method comprises selecting a peptide immunogen of the target protein by the above-described method, synthesizing the peptide immunogen, conjugating the peptide immunogen to an immunogenic carrier molecule to make a peptide-carrier antigen, and producing antibodies to the antigen.
- the present invention is directed to an antibody made by the above-described method. Furthermore, the present invention is directed to a peptide which consists of a sequence from a tissue factor extracellular region from a first species which is 5-50 amino acids in length, has a hydrophilicity value greater than 0, and has at least 1 nonhomologous amino acid from a tissue factor from a second species .
- the present invention is also directed to an antigen, and antibodies made to an antigen, where the antigen comprises an immunogenic carrier molecule conjugated to a peptide which consists of a sequence from a tissue factor extracellular region from a first species which is 5-50 amino acids in length, has a hydrophilicity value greater than 0, and has at least 1 nonhomologous amino acid from a tissue factor from a second species .
- TF shall mean tissue factor.
- a protein from a first species but not the same protein from a second species is said to be species-specific to the protein from the first species, because it does not "cross- react” to the protein from the second species.
- synthetic peptide or “peptide” is a chain of 5 to 50 amino acids covalently linked to one another by peptide bonds.
- an "antigen” is a macromolecule to which antibodies are made.
- the antigen is usually injected into a vertebrate, which then elicits production of antibodies to regions (“epitopes”) of the antigen.
- immunogen is a portion of an antigen to which antibodies are desired.
- An immunogen may be an epitope of the antigen or it may be comprised of several epitopes. More specifically, the term “immunogen” is applied herein to mean a peptide from a protein of interest which is conjugated to an immunogenic carrier molecule for the purpose of eliciting antibodies to the immunogen portion of the antigen.
- hydrophilic when used in reference to amino acids refers to those amino acids which have polar and/or charged side chains. Hydrophilic amino acids include lysine, arginine, histidine, aspartate (i.e., aspartic acid), glutamate (i.e., glutamic acid), serine, threonine, cysteine, tyrosine, asparagine and glutamine.
- hydrophilicity value or "Hydrophilicity index value” as used herein means a number value assigned to each amino acid from 3.0 to -3.4, where the larger numbers are more hydrophilic. The hydrophilicity values are calculated using the method of Hopp et al . , 1981, Proc.
- hydrophobic when used in reference to amino acids refers to those amino acids which have nonpolar side chains. Hydrophobic amino acids include valine, leucine, isoleucine, cysteine and methionine. Three hydrophobic amino acids have aromatic side chains. Accordingly, the term "aromatic” when used in reference to amino acids refers to the three aromatic hydrophobic amino acids phenylalanine, tyrosine and tryptophan.
- FIGURE 1 is a sequence homology comparison of the rabbit, human, and mouse tissue factor proteins. The figure was taken from Pawashe et al . , 1991, Thromb. Haemost . 66:315.
- FIGURE 2 is a list of candidate immunogenic peptides from rabbit tissue factor selected using the methods of the present invention.
- FIGURE 3 is a list of candidate immunogenic peptides from human tissue factor selected using the methods of the present invention. DETAILED DESCRIPTION OF THE INVENTION
- the present invention is directed to methods of selecting a peptide immunogen consisting of a region of a target protein from a first species, where the target protein has a different amino acid sequence in a second species.
- the immunogen is selected for the purpose of eliciting antibodies to the region of the target protein containing the peptide.
- the method comprises the following steps: (a) the amino acid sequence of the target protein is analyzed for hydrophilicity, and regions from 5-50 amino acids long are identified which are hydrophilic, for example having a value greater than 0 using the method of Hopp et al., 1981, Proc. Natl. Acad. Sci. U.S.A.
- the hydrophilic amino acid regions of the target protein are compared to the same regions from the same protein of the second species, and a region is selected as the peptide immunogen which has at least 1 nonhomologous amino acid between the first species and the second species.
- Antibodies are then made to the selected immunogen after conjugating the immunogen to an immunogenic carrier molecule.
- Antibodies which are generated to the immunogen generally do not bind to the same protein of the second species.
- the method of the current invention may be applied for the production of antibodies to any target protein in a species of interest to which there exists an analogous protein in another species which has an amino acid sequence which is different from the amino acid sequence in the target protein in the species of interest.
- proteins from any prokaryotic, eukaryotic, or viral source including but not limited to, vertebrates, invertebrates, protozoans, bacteria, fungi, and plants .
- the amino acid sequences of the target protein and the analogous protein from the second species can be obtained from various sources, such as published material, as well as protein sequencing (e.g. Edman degradation).
- the amino acid sequences may also be deduced from the genes encoding the target proteins which can be obtained from published sources or by cloning the genes encoding the target proteins from both species and determining the start codon by known methods.
- hydrophilic peptide regions are utilized because those regions tend to be more antigenic than hydrophobic regions. Hydrophilic peptide regions also tend to localize on the outer surface of the target protein, and are therefore more accessible for eliciting an antibody response and for being exposed to an antibody in in si tu studies such as immunocytochemical investigations.
- Using hydrophilic peptides to produce immunogens also have the advantage of being more soluble in aqueous solution than hydrophobic peptides. This makes the procedure for conjugating the peptides to carrier proteins simpler with preferred conjugation procedures. Additionally, antigens comprising hydrophilic peptides are more soluble than antigens comprising hydrophobic peptides, making immunization procedures with the former antigen simpler.
- a hydrophilicity analysis provides a determination of the moving average of the polar portion, preferably a hexapeptide, of the protein.
- the hydrophilic epitopes of tissue factor may be determined by the method of Hopp et al . , 1981, Proc. Natl. Acad. Sci. U.S.A. 76 . : 3824.
- Computer software such as MacVector from International Biotechnologies, Inc., are also available to analyze hydrophilicity of a target protein.
- hydrophilicity values for individual amino acids range from -3.4 (for tryptophan) to 3.0 (for lysine, glutamic acid, aspartic acid, and arginine)
- the hydrophilicity is preferably determined by averaging the mean hexapeptide hydrophilicity value over the length of the peptide.
- Peptides may be selected which have a hydrophilicity greater than 0.
- Preferred peptides have a hydrophilicity greater than 0.2.
- the sequence of each peptide is compared with the analogous sequence in the same protein from the second species.
- the amino acid sequences of the target protein and the same protein from the second species are preferably aligned by 3 -dimensional structural similarity.
- the structural similarity between the two sequences can be determined by comparing predicted 3 -dimensional structure which can be determined by a number of computer programs, or by determining the crystal structure of both of the proteins by known methods (see, e.g., Harlos et al . , 1994, Nature 370 :662 for the crystal structure of TF) .
- 3 -dimensional structural similarity can generally be ascertained by aligning the sequences of the two proteins by introducing gaps in one or the other sequence to minimize mismatches.
- This task can also be performed by a number of well-known computer programs (see, e.g., Altschul et al., 1997, Nucl . Acids Res. 25_:3389). See Figure 1 for such an alignment of mouse, rabbit, and human TF .
- the aligned sequences of the candidate hydrophilic peptides are compared for sequence homology.
- Peptides which have at least 1 nonhomologous amino acid between the target protein and the analogous protein from the second species may be used as immunogens according to the present invention.
- the candidate hydrophilic peptides have at least 3 nonhomologous amino acids between the target protein and the analogous protein from the second species.
- the nonhomologous amino acids of the candidate hydrophilic peptides may or may not be consecutive in the peptide chain.
- the homology between the two peptide sequences is less than 75%. Even more preferably the homology between the two peptide sequences is less than 60%.
- the peptide immunogen is selected, it is synthesized in preparation for conjugation to the carrier molecule. Synthesis of the peptide can be by any convenient method, for example by translating a nucleic acid sequence encoding the peptide in a suitable recombinant host cell, e.g., E. coli , then purifying the peptide by routine methods (e.g., reversed phase HPLC) . However, the peptide is preferably synthesized using chemical methods, preferably solid phase methods. The preferred method for this employs Fmoc chemistry (Fields et al . , 1990, Int. J. Pept. Protein Res. 35:161; Wellings et al . , 1997, Meth.
- the peptide is selected with a terminal amino acid residue suitable for coupling to the carrier with a crosslinking reagent, or synthesized with an additional amino acid residue at the terminus for this purpose.
- amino acids examples include tyrosine, for a bis-diazobenzidine (BDB) - activated carrier; aspartic acid, glutamic acid, or a free carboxy terminus, for a l-ethyl-3- (3 -dimethylaminopropyl) carbodiimide hydrochloride (EDC) -activated carrier; or cysteine, lysine, or a free amino terminus, for a m-maleimidobenzoyl-N- hydrosuccinamide (MBS) -activated carrier.
- BDB bis-diazobenzidine
- EDC l-ethyl-3- (3 -dimethylaminopropyl) carbodiimide hydrochloride
- cysteine, lysine, or a free amino terminus for a m-maleimidobenzoyl-N- hydrosuccinamide (MBS) -activated carrier.
- cysteine and MBS carrier activation Preferred is cyst
- the carrier is preferably an antigenic, soluble protein.
- the carrier is selected which would not elicit antibodies which would bind to antigens in the first or second species.
- a preferred carrier is keyhole limpet hemocyanin (KLH) .
- the peptide immunogen can be conjugated to a resin-polylysine carrier, to follow the multiple antigenic peptide strategy. See, e.g., McClean et al.,1991, J. Immunol. Meth. 137:149.
- Production of the antigen is achieved by conjugating the peptide immunogen to the carrier molecule.
- This conjugation preferably proceeds by activation of the carrier, e.g., by BDB, EDC, or, preferably, MBS, then addition of the peptide.
- the antigen is then used to produce an antibody which binds to the immunogen.
- the antibody may be a mono-specific antibody.
- the monospecific antibody may be a monoclonal antibody produced, for example, by the method of Galfre et al . , 1977, Nature 266:550.
- the monospecific antibody may be a recombinant antibody produced, for example, by the method of Lowman et al . , 1991, Biochemistry 30:10832.
- the antibody can also be a polyclonal antibody.
- the polyclonal antibody can be prepared, for example, by immunizing a mammal such as a mouse, goat, sheep, or rabbit with the peptide- carrier antigen and subsequently isolating the serum therefrom to obtain an antiserum that contains the polyclonal antibodies.
- the antibody is preferably characterized by determining its ability to bind to the immunogen and the target protein by, e.g., ELISA, hemagglutination, fluorescent antibody binding to tissue, western blot, or any other suitable procedure.
- Tissue factor antibodies made by the invention method can be tested, e.g., for reactivity to the immunogen, and to recombinant and/or native TF from the first and second species, or other species.
- the peptide antigens, immunogens, and antibodies of the present invention can be used in various applications. Among other uses, the peptide antigens can be employed to purify the antibodies as noted above.
- the synthetic peptides of the present invention can be used to test antibody specificity as noted above.
- the antigens can also be used in inhibition assays. For example, a peptide from TF can be added to, e.g., thromboplatin assays to determine if it can inhibit TF function. Such work could help characterize TF structure and function.
- antibodies of the present invention include epitope mapping in structure/function studies, immunohistochemical localization, and immunoaffinity purification of the target protein.
- anti-TF antibodies can be used in a thromboplastin assays to determine if the antibody can inhibit TF function. These antibodies can also be used to study other aspects of TF, such as characteristics of the TF/substrate complex.
- the antibodies of the present invention can also be used in diagnostic assays for antigens, particularly where it is required that the antibodies do not cross-react with antigens from another species (e.g., assays for a particular bacteria in food, etc.)
- Tissue Factor The sequences of rabbit, human, and mouse tissue factor were have been described (Pawashe et al . , 1991, Thromb Haemost . 6_6:315) . A hydrophilicity analysis was performed using the method of Hopp et al . , 1981, Proc. Natl. Acad. Sci. U.S.A.
- the candidate immunogens were next evaluated for homology to the mouse sequence.
- the human and rabbit sequence was aligned with the mouse sequence as in Figure 1.
- the candidate human and rabbit immunogens were then compared to the aligned mouse sequence and immunogens were selected which have more than 3 nonhomologous amino acids from the analogous mouse sequence.
- the selected immunogens are shown as the candidate immunogens of Figures 2 and 3 which have 3 or more mismatches. Eight of the ten candidate immunogens from rabbit TF and eight of the eleven candidate immunogens from human TF . All of those immunogens are useful for producing anti-TF antibodies. However, the most effective immunogens would be those which have a higher mean hydrophilicity value and a larger number of mismatches.
- Rabbit Tissue Factor Polyclonal antibodies were made to two regions of rabbit TF .
- the 12 residue peptide TTGFPEEPPFRN from position 84 through 95 (84T-95S) located in the loop connecting domains IF and IG and oriented toward the putative top of the molecule (Harlos et al . , 1994, Nature 370 : 662) , demonstrated the most amino acids mismatches of the candidate immunogens ( Figure 2) .
- the 13 residue peptide VIPSRKRKQRSPE from position 190 through 202 (190V-202E) , located within the 2F-2G loop constrained by a disulfide bond, had the highest hydrophilicity index (Figure 2) .
- Both peptides are unique to the rabbit TF sequence when analyzed by the BLASTP search engine with an E value of 100. These two peptides were synthesized using standard FMOC chemistry. An additional cysteine was added to the carboxy terminal. The peptides were then conjugated to MBS-activated KLH by standard methods. For each peptide immunogen, three goats were immunized with the peptide-KLH antigens. Antisera collected from the goats were then tested for immunoreactivity in a microtiter plate format in which unconjugated peptides, rabbit recombinant TF, human recombinant TF, and a Triton extract of rabbit brain acetone powder (RBAP) were the coated antigens.
- RBAP Triton extract of rabbit brain acetone powder
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Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AU63384/00A AU755574B2 (en) | 1999-06-30 | 2000-06-27 | Synthetic peptide immunogens and antibodies thereto |
EP00950257A EP1105158A1 (en) | 1999-06-30 | 2000-06-27 | Synthetic peptide immunogens and antibodies thereto |
JP2001505944A JP2003503040A (en) | 1999-06-30 | 2000-06-27 | Synthetic peptide immunogens and antibodies thereto |
CA002339404A CA2339404A1 (en) | 1999-06-30 | 2000-06-27 | Synthetic peptide immunogens and antibodies thereto |
Applications Claiming Priority (2)
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US14169999P | 1999-06-30 | 1999-06-30 | |
US60/141,699 | 1999-06-30 |
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WO2001000237A1 true WO2001000237A1 (en) | 2001-01-04 |
WO2001000237A9 WO2001000237A9 (en) | 2002-05-02 |
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PCT/US2000/017609 WO2001000237A1 (en) | 1999-06-30 | 2000-06-27 | Synthetic peptide immunogens and antibodies thereto |
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EP (1) | EP1105158A1 (en) |
JP (1) | JP2003503040A (en) |
AU (1) | AU755574B2 (en) |
CA (1) | CA2339404A1 (en) |
WO (1) | WO2001000237A1 (en) |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2001075086A2 (en) * | 2000-03-31 | 2001-10-11 | Zymogenetics, Inc. | Multi-domain proteinase inhibitor |
US7267960B2 (en) | 2003-07-25 | 2007-09-11 | Amgen Inc. | Antagonists and agonists of LDCAM and methods of use |
US7402655B2 (en) | 1998-08-07 | 2008-07-22 | Immunex Corporation | Molecules designated LDCAM |
WO2009067272A1 (en) * | 2007-11-19 | 2009-05-28 | Abbott Laboratories | Immunoassay for detection and quantification of amyloid-b peptides |
US9580544B2 (en) | 2010-03-18 | 2017-02-28 | New Jersey Institute Of Technology | Polyesters from asymetrical monomers based upon bisanhydrohexitols |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP4597172B2 (en) * | 2007-08-29 | 2010-12-15 | 丸大食品株式会社 | Antibody to bovine myoglobin partial peptide, test method using the antibody, and test kit |
-
2000
- 2000-06-27 JP JP2001505944A patent/JP2003503040A/en active Pending
- 2000-06-27 CA CA002339404A patent/CA2339404A1/en not_active Abandoned
- 2000-06-27 EP EP00950257A patent/EP1105158A1/en not_active Withdrawn
- 2000-06-27 WO PCT/US2000/017609 patent/WO2001000237A1/en active Application Filing
- 2000-06-27 AU AU63384/00A patent/AU755574B2/en not_active Ceased
Non-Patent Citations (2)
Title |
---|
HARLOS ET AL.: "Crystal structure of the extracellular region of human tissue factor", NATURE, vol. 370, 25 August 1994 (1994-08-25), pages 662 - 666, XP002932454 * |
HOPP T.P.: "Use of hydrophilicity plotting procedures to identify protein antigenic segments and other interaction sites", METHODS IN ENZYMOLOGY, vol. 178, 1989, pages 571 - 585, XP002932455 * |
Cited By (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7402655B2 (en) | 1998-08-07 | 2008-07-22 | Immunex Corporation | Molecules designated LDCAM |
US7659385B2 (en) | 1998-08-07 | 2010-02-09 | Immunex Corporation | Polynucleotides encoding molecules designated LDCAM |
US7741115B2 (en) | 1998-08-07 | 2010-06-22 | Immunex Corporation | Antibodies that bind LDCAM |
WO2001075086A2 (en) * | 2000-03-31 | 2001-10-11 | Zymogenetics, Inc. | Multi-domain proteinase inhibitor |
WO2001075086A3 (en) * | 2000-03-31 | 2002-06-27 | Zymogenetics Inc | Multi-domain proteinase inhibitor |
US7267960B2 (en) | 2003-07-25 | 2007-09-11 | Amgen Inc. | Antagonists and agonists of LDCAM and methods of use |
US8043615B2 (en) | 2003-07-25 | 2011-10-25 | Amgen Inc. | Methods of antagonizing LDCAM and CRTAM |
WO2009067272A1 (en) * | 2007-11-19 | 2009-05-28 | Abbott Laboratories | Immunoassay for detection and quantification of amyloid-b peptides |
US9580544B2 (en) | 2010-03-18 | 2017-02-28 | New Jersey Institute Of Technology | Polyesters from asymetrical monomers based upon bisanhydrohexitols |
US10364251B2 (en) | 2010-03-18 | 2019-07-30 | New Jersey Institute Of Technology | Polyesters from assymetrical monomers based upon bisanhydrohexitols |
Also Published As
Publication number | Publication date |
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CA2339404A1 (en) | 2001-01-04 |
EP1105158A1 (en) | 2001-06-13 |
AU755574B2 (en) | 2002-12-19 |
JP2003503040A (en) | 2003-01-28 |
AU6338400A (en) | 2001-01-31 |
WO2001000237A9 (en) | 2002-05-02 |
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