WO2007062852A2 - ANTI-Aβ GLOBULOMER ANTIBODIES, ANTIGEN-BINDING MOIETIES THEREOF, CORRESPONDING HYBRIDOMAS, NUCLEIC ACIDS, VECTORS, HOST CELLS, METHODS OF PRODUCING SAID ANTIBODIES, COMPOSITIONS COMPRISING SAID ANTIBODIES, USES OF SAID ANTIBODIES AND METHODS OF USING SAID ANTIBODIES - Google Patents
ANTI-Aβ GLOBULOMER ANTIBODIES, ANTIGEN-BINDING MOIETIES THEREOF, CORRESPONDING HYBRIDOMAS, NUCLEIC ACIDS, VECTORS, HOST CELLS, METHODS OF PRODUCING SAID ANTIBODIES, COMPOSITIONS COMPRISING SAID ANTIBODIES, USES OF SAID ANTIBODIES AND METHODS OF USING SAID ANTIBODIES Download PDFInfo
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- WO2007062852A2 WO2007062852A2 PCT/EP2006/011530 EP2006011530W WO2007062852A2 WO 2007062852 A2 WO2007062852 A2 WO 2007062852A2 EP 2006011530 W EP2006011530 W EP 2006011530W WO 2007062852 A2 WO2007062852 A2 WO 2007062852A2
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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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- 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
-
- 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
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
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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
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4711—Alzheimer's disease; Amyloid plaque core protein
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
- G01N33/6896—Neurological disorders, e.g. Alzheimer's disease
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/40—Immunoglobulins specific features characterized by post-translational modification
- C07K2317/41—Glycosylation, sialylation, or fucosylation
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
- C07K2317/565—Complementarity determining region [CDR]
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
- C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
- G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
- G01N2333/4701—Details
- G01N2333/4709—Amyloid plaque core protein
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/28—Neurological disorders
- G01N2800/2814—Dementia; Cognitive disorders
- G01N2800/2821—Alzheimer
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/38—Pediatrics
- G01N2800/385—Congenital anomalies
- G01N2800/387—Down syndrome; Trisomy 18; Trisomy 13
Definitions
- the present invention relates to anti-A ⁇ globulomer antibodies, antigen-binding moieties thereof, hybridomas producing said antibodies, nucleic acids encoding said antibodies, vectors comprising said nucleic acids, host cells comprising said vectors, methods of producing said antibodies, compositions comprising said antibodies, therapeutic and diagnostic uses of said antibodies and corresponding methods relating to Alzheimer's disease and other amyloidoses.
- Alzheimer's disease is the most frequent cause for dementia among the aged with an incidence of about 10% of the population above 65 years. With increasing age, the probability of disease also rises. Globally, there are about 15 million people affected and further increases in life expectancy are expected to increase the number of diseased people to about threefold over the next decades.
- AD Alzheimer's disease
- a ⁇ amyloid ⁇
- a ⁇ amyloid ⁇
- the amyloid ⁇ (A ⁇ ) peptide arises from the ⁇ -amyloid precursor protein by proteolytic cleavage. This cleavage is effected by the cooperative activity of several proteases named ⁇ -, ⁇ - and ⁇ -secretase. Cleavage leads to a number of specific fragments of differing length.
- the amyloid plaques consist mostly of peptides with a length of 40 or 42 amino acids (A ⁇ 40, A ⁇ 42). The dominant cleavage product is A ⁇ 40; however, A ⁇ 42 has a much stronger toxic effect.
- Cerebral amyloid deposits and cognitive impairments very similar to those observed in AIz- heimer's disease are also hallmarks of Down's syndrome (trisomy 21 ), which occurs at a frequency of about 1 in 800 births.
- Meningoencephalitis was characterized by neuroinflammation and infiltration of T-cells into the brain. Presumably, this was due to a T-cell immune response induced by injection of A ⁇ (1-42) as antigen. Such an immune response is not to be expected after passive immunization. To date, there are no clinical data with reference to this available yet. However, with reference to such a passive approach to immunization concerns about the side effect profile were voiced because of preclinical studies in very old APP23 mice which received an antibody directed against an N-terminal epitope of A ⁇ (1-42) once a week over 5 months.
- mice showed an increase in the number and severity of microhaemorrhages compared to control animals treated with saline (Pfeifer et al., 2002, Science, 298, 1379).
- a comparable increase in micro- haemorrhages was also described in very old (> 24 months) Tg2576 and PDAPP mice (Racke et al., 2005, J Neurosci, 25, 629-636; Wilcock et al. 2004, J. Neuroinflammation, 1(1 ):24; De Mattos et al., 2004, Neurobiol. Aging 25(S2):577).
- antibody injection led to a significant increase in microhaemorrhages.
- WO2004/067561 describes stable A ⁇ (1 -42) oligomers (A ⁇ (1-42) globulomers) and antibodies directed specifically against the globulomers. Digestion with unspecific proteases shows that the A ⁇ globulomer may be digested beginning with the hydrophilic N-terminus protruding from the globular core structure (Barghom et al., 2005, J Neurochem, 95, 834-847). Such N- terminal truncated A ⁇ globulomers (A ⁇ (12-42) and A ⁇ (20-42) globulomers) represent the basic structural unit of this oligomeric A ⁇ .
- a therapeutically questionable reduction of brain volume has been observed in the study of active immunization with A ⁇ peptides in fibrillary condition of aggregation (ELAN trial with AN1792). Moreover, in this trial severe side effects in form of a meningoencephalitis were observed.
- the present invention solves this problem by providing globulomer-specific antibodies possessing high affinity for truncated forms of A ⁇ globulomers. These antibodies are capable of discriminating not only other forms of A ⁇ peptides, particularly monomers and fibrils, but also untruncated forms of A ⁇ globulomers.
- the present invention relates to an antibody having a binding affinity to an A ⁇ (20-42) globulomer that is greater than the binding affinity of this antibody to an A ⁇ (1 -42) globulomer.
- the present invention relates to an antibody having a binding affinity to an A ⁇ (20-42) globulomer that is greater than the binding affinity of this antibody to an A ⁇ (12-42) globulomer.
- the invention thus relates to antibodies having a binding affinity to the A ⁇ (20-42) globulomer that is greater than the binding affinity of the antibody to both the A ⁇ (1-42) globulomer and the A ⁇ (12-42) globulomer.
- a ⁇ (X-Y) here refers to the amino acid sequence from amino acid position X to amino acid position Y of the human amyloid ⁇ protein including both X and Y, in particular to the amino acid sequence from amino acid position X to amino acid position Y of the amino acid sequence DAEFRHDSGY EVHHQKLVFF AEDVGSNKGA IIGLMVGGVV IAT (corresponding to amino acid positions 1 to 43) or any of its naturally occurring variants, in particular those with at least one mutation selected from the group consisting of A2T, H6R, D7N, A21G ("Flemish"), E22G (“Arctic”), E22Q (“Dutch”), E22K (“Italian”), D23N (“Iowa”), A42T and A42V wherein the numbers are relative to the start of the A ⁇ peptide, including both position X and position Y or a sequence with up to three additional amino acid substitutions none of which may prevent globulomer formation, preferably with no
- a ⁇ (1-42) here refers to the amino acid sequence from amino acid position 1 to amino acid position 42 of the human amyloid ⁇ protein including both 1 and 42, in particular to the amino acid sequence DAEFRHDSGY EVHHQKL VFF AEDVGSNKGA IIGLMVGGW IA or any of its naturally occurring variants, in particular those with at least one mutation selected from the group consisting of A2T, H6R, D7N, A21G ("Flemish”), E22G (“Arctic”), E22Q (“Dutch”), E22K (“Italian”), D23N (“Iowa”), A42T and A42V wherein the numbers are relative to the start of the A ⁇ peptide, including both 1 and 42 or a sequence with up to three additional amino acid substitutions none of which may prevent globulomer formation, preferably with no additional amino acid substitutions in the portion from amino acid 20 to amino acid 42.
- a ⁇ (1-40) here refers to the amino acid sequence from amino acid position 1 to amino acid position 40 of the human amyloid ⁇ protein including both 1 and 40, in particular to the amino acid sequence DAEFRHDSGY EVHHQKLVFF
- a ⁇ (12-42) here refers to the amino acid sequence from amino acid position 12 to amino acid position 42 of the human amyloid ⁇ protein including both 12 and 42, in particular to the amino acid sequence VHHQKLVFF AEDVGSNKGA IIGLMVGGW IA or any of its naturally occurring variants, in particular those with at least one mutation selected from the group consisting of A21 G ("Flemish”), E22G (“Arctic”), E22Q (“Dutch”), E22K (“Italian”), D23N (“Iowa”), A42T and A42V wherein the numbers are relative to the start of the A ⁇ peptide, including both 12 and 42 or a sequence with up to three additional amino acid substitutions none of which may prevent globulomer formation, preferably with no additional amino acid substitutions in the portion from amino acid 20 to amino acid 42.
- a ⁇ (20-42) here refers to the amino acid sequence from amino acid position 20 to amino acid position 42 of the human amyloid ⁇ protein including both 20 and 42, in particular to the amino acid sequence F AEDVGSNKGA IIGLMVGGW IA or any of its naturally occurring variants, in particular those with at least one mutation selected from the group consisting of A21G ("Flemish”), E22G (“Arctic”), E22Q (“Dutch”), E22K (“Italian”), D23N (“Iowa”), A42T and A42V wherein the numbers are relative to the start of the A ⁇ peptide, in- eluding both 20 and 42 or a sequence with up to three additional amino acid substitutions none of which may prevent globulomer formation, preferably without any additional amino acid substitutions.
- a ⁇ (X-Y) globulomer refers to a soluble, globular, non-covalent association of A ⁇ (X-Y) peptides as defined above, possessing homogeneity and distinct physical characteristics.
- a ⁇ (X-Y) globulomers are stable, non-fibrillar, oligomeric assemblies of A ⁇ (X-Y) peptides which are obtainable by incubation with anionic detergents. In contrast to monomer and fibrils, these globulomers are characterized by defined assembly numbers of subunits (e.g.
- the globulomers have a 3-dimensional globular type structure ("molten globule”, see Barghorn et al., 2005, J Neurochem, 95, 834-847). They may be further characterized by one or more of the following features:
- a ⁇ (X-Y) globulomer here refers in particular to a product which is obtainable by a process as described in WO 2004/067561 , which is incorporated herein by reference.
- Said process comprises unfolding a natural, recombinant or synthetic A ⁇ (X-Y) peptide or a derivative thereof; exposing the at least partially unfolded A ⁇ (X-Y) peptide or derivative thereof to a detergent, reducing the detergent action and continuing incubation.
- hydrogen bond-breaking agents such as, for exam- pie, hexafluoroisopropanol (HFIP) may be allowed to act on the protein. Times of action of a few minutes, for example about 10 to 60 minutes, are sufficient when the temperature of action is from about 20 to 5O 0 C and in particular about 35 to 4O 0 C. Subsequent dissolution of the residue evaporated to dryness, preferably in concentrated form, in suitable organic solvents miscible with aqueous buffers, such as, for example, dimethyl sulfoxide (DMSO), results in a suspension of the at least partially unfolded peptide or derivative thereof, which can be used subsequently. If required, the stock suspension may be stored at low temperature, for example at about -20 0 C, for an interim period.
- DMSO dimethyl sulfoxide
- the peptide or the derivative thereof may be taken up in slightly acidic, preferably aqueous, solution, for example an about 10 mM aqueous HCI solution.
- aqueous solution for example an about 10 mM aqueous HCI solution.
- insoluble components are removed by centrifugation. A few minutes at 10000 g is expedient.
- These method steps are preferably carried out at room temperature, i.e. a temperature in the range from 20 to 3O 0 C.
- the supernatant obtained after centrifugation contains the A ⁇ (X-Y) peptide or the derivative thereof and may be stored at low temperature, for example at about -20°C, for an interim period.
- oligomers A an intermediate type of oligomers (in WO 2004/067561 referred to as oligomers A).
- a detergent is allowed to act on the at least partially unfolded peptide or derivative thereof until sufficient intermediate oligomer has been produced.
- ionic detergents in particular anionic detergents.
- R-X is used, in which the radical R is unbranched or branched alkyl having from 6 to 20 and preferably 10 to 14 carbon atoms or unbranched or branched alkenyl having from 6 to 20 and preferably 10 to 14 carbon atoms, the radical X is an acidic group or salt thereof, with X being preferably selected from among -COO-M + , -SO 3 -M + , and especially
- -OSO 3 -M + and M + is a hydrogen cation or an inorganic or organic cation preferably selected from alkali metal and alkaline earth metal cations and ammonium cations.
- Tetaine acid and oleic acid can also be used advantageously.
- the sodium salt of the detergent lauroylsarcosin also known as sarkosyl NL-30 or Gardol ® ) is also particularly advantageous.
- the time of detergent action in particular depends on whether - and if yes, to what extent - the peptide or the derivative thereof subjected to oligomerization has unfolded. If 1 according to the unfolding step, the peptide or derivative thereof has been treated beforehand with a hydrogen bond-breaking agent, i.e. in particular with hexafluoroisopropanol, times of action in the range of a few hours, advantageously from about 1 to 20 and in particular from about 2 to 10 hours, are sufficient when the temperature of action is about 20 to 5O 0 C and in particular about 35 to 40 0 C. If a less unfolded or an essentially not unfolded peptide or derivative thereof is the start- ing point, correspondingly longer times of action are expedient.
- a hydrogen bond-breaking agent i.e. in particular with hexafluoroisopropanol
- times of action in the range from about 5 to 30 hours and in particular from about 10 to 20 hours are sufficient when the temperature of action is about 20 to 50°C and in particular about 35 to 4O 0 C.
- insoluble components are advantageously removed by centrifugation. A few minutes at 10000 g is expedient.
- the detergent concentration to be chosen depends on the detergent used. If SDS is used, a concentration in the range from 0.01 to 1% by weight, preferably from 0.05 to 0.5% by weight, for example of about 0.2% by weight, proves expedient. If lauric acid or oleic acid are used, somewhat higher concentrations are expedient, for example in a range from 0.05 to 2% by weight, preferably from 0.1 to 0.5% by weight, for example of about 0.5% by weight.
- the detergent action should take place at a salt concentration approximately in the physiologi- cal range.
- NaCI concentrations in the range from 50 to 500 mM, preferably from 100 to 200 mM and particularly at about 140 mM are expedient.
- oligomers B The subsequent reduction of the detergent action and continuation of incubation relates to a further oligomerization to give the A ⁇ (X-Y) globulomer of the invention (in WO 2004/067561 referred to as oligomers B).
- the composition obtained from the preceding step regularly contains detergent and a salt concentration in the physiological range it is then expedient to reduce detergent action and, preferably, also the salt concentration. This may be carried out by reducing the concentration of detergent and salt, for example, by diluting, expediently with water or a buffer of lower salt concentration, for example Tris-HCI, pH 7.3. Dilution factors in the range from about 2 to 10, advantageously in the range from about 3 to 8 and in particular of about 4, have proved suitable.
- the reduction in detergent action may also be achieved by adding substances which can neutralize said detergent action.
- substances which can neutralize said detergent action include substances capable of complexing the detergents, like substances capable of stabilizing cells in the course of purification and extraction measures, for example particular EO/PO block co- polymers, in particular the block copolymer under the trade name Pluronic ® F 68.
- Alkoxylated and, in particular, ethoxylated alkyl phenols such as the ethoxylated t-octylphenols of the Triton ® X series, in particular Triton ® X100, 3-(3-cholamidopropyldimethylammonio)-1- propanesulfonate (CHAPS ® ) or alkoxylated and, in particular, ethoxylated sorbitan fatty esters such as those of the Tween ® series, in particular Tween ® 20, in concentration ranges around or above the particular critical micelle concentration, may be equally used.
- ethoxylated alkyl phenols such as the ethoxylated t-octylphenols of the Triton ® X series, in particular Triton ® X100, 3-(3-cholamidopropyldimethylammonio)-1- propanesulfonate (CHAPS ® ) or alkoxylated
- the solution is incubated until sufficient A ⁇ (X-Y) globulomer of the invention has been produced. Times of action in the range of several hours, preferably in the range from about 10 to 30 hours and in particular in the range from about 15 to 25 hours, are sufficient when the temperature of action is about 20 to 50 0 C and in particular about 35 to 40 0 C.
- the solution may then be concentrated and possible residues may be removed by centrifugation. Here too, a few minutes at 10000 g proves expedient.
- the supernatant obtained after centrifugation contains an A ⁇ (X-Y) globulomer of the invention.
- An A ⁇ (X-Y) globulomer of the invention can be finally recovered in a manner known per se, e. g. by ultrafiltration, dialysis, precipitation or centrifugation.
- electrophoretic separation of the A ⁇ (X-Y) globulomers under denaturing conditions produces a double band (e. g. with an apparent molecular weight of 38 / 48 kDa for A ⁇ (1-42)), and especially preferred if upon glutardialdehyde treatment of the globulomers before separation these two bands are merged into one.
- size exclusion chromatography of the globulomers results in a single peak (e. g. corresponding to a molecular weight of approximately 100 kDa for A ⁇ (1-42) globulomer or of approximately 60 kDa for glutardialdehyde cross-linked A ⁇ (1-42) globulomer), respectively.
- a ⁇ (X-Y) globulomers wherein X is selected from the group consisting of the numbers 2 .. 24 and Y is as defined above are those which are obtainable by truncating A ⁇ (1-Y) globulomers into shorter forms wherein X is selected from the group consisting of the numbers 2 .. 24, with X preferably being 20 or 12, and Y is as defined above, which can be achieved by treatment with appropriate proteases.
- an A ⁇ (20-42) globulomer can be obtained by subjecting an A ⁇ (1-42) globulomer to thermolysin proteolysis
- an A ⁇ (12-42) globulomer can be obtained by subjecting an A ⁇ (1-42) globulomer to endoproteinase GIuC proteolysis.
- the protease is inactivated in a generally known manner.
- the resulting globulomers may then be isolated following the procedures already described herein and, if required, processed fur- ther by further work-up and purification steps. A detailed description of said processes is dis- closed in WO 2004/067561 , which is incorporated herein by reference.
- an A ⁇ (1-42) globulomer is in particular the A ⁇ (1-42) globulomer as described in example 1a herein; an A ⁇ (20-42) globulomer is in particular the A ⁇ (20-42) globulomer as described in examples 1c herein, and an A ⁇ (12-42) globulomer is in particular the A ⁇ (12-42) globulomer as described in examples 1d herein.
- the globulomer shows affinity to neuronal cells.
- the globulomer also exhibits neuromodulating effects.
- the globulomer consists of 11 to 16, and most preferably, of 12 to 14 A ⁇ (X-Y) peptides.
- the term "A ⁇ (X-Y) globulomer” here refers to a globulomer consisting essentially of A ⁇ (X-Y) subunits, where it is preferred if on average at least 11 of 12 subunits are of the A ⁇ (X-Y) type, more preferred if less than 10% of the globu- lomers comprise any non-A ⁇ (X-Y) peptides, and most preferred if the content of non-A ⁇ (X-Y) peptides is below the detection threshold.
- a ⁇ (1-42) globulomer here refers to a globulomer consisting essentially of A ⁇ (1-42) units as defined above
- a ⁇ (12-42) globulomer here refers to a globulomer consisting essentially of A ⁇ (12-42) units as defined above
- a ⁇ (20- 42) globulomer here refers to a globulomer consisting essentially of A ⁇ (20-42) units as defined above.
- cross-linked A ⁇ (X-Y) globulomer here refers to a molecule obtainable from an A ⁇ (X-Y) globulomer as described above by cross-linking, preferably chemically cross-linking, more preferably aldehyde cross-linking, most preferably glutardialdehyde cross-linking of the constituent units of the globulomer.
- a cross-linked globu- lomer is essentially a globulomer in which the units are at least partially joined by covalent bonds, rather than being held together by non-covalent interactions only.
- a cross-linked A ⁇ (1-42) globulomer is in particular the cross-linked A ⁇ (1- 42) oligomer as described in example 1 b herein.
- a ⁇ (X-Y) globulomer derivative here refers in particular to a globulomer that is labelled by being covalently linked to a group that facilitates detection, preferably a fluorophore, e. g. fluorescein isothiocyanate, phycoerythrin, Aequorea victoria fluorescent protein, Dic- tyosoma fluorescent protein or any combination or fluorescence-active derivative thereof; a chromophore; a chemoluminophore, e. g.
- a fluorophore e. g. fluorescein isothiocyanate, phycoerythrin, Aequorea victoria fluorescent protein, Dic- tyosoma fluorescent protein or any combination or fluorescence-active derivative thereof
- a chromophore e. a chemoluminophore, e. g.
- luciferase preferably Photinus pyralis luciferase, Vibrio fischeri luciferase, or any combination or chemoluminescence-active derivative thereof; an enzymatically active group, e. g. peroxidase, e. g. horseradish peroxidase, or any enzy- matically active derivative thereof; an electron-dense group, e. g. a heavy metal containing group, e.g. a gold containing group; a hapten, e. g. a phenol derived hapten; a strongly antigenic structure, e. g. peptide sequence predicted to be antigenic, e. g.
- a chelating group e. g. hexahistidinyl
- a natural or nature-derived protein structure mediating further specific protein-protein interactions, e. g. a member of the fos/jun pair
- a magnetic group e. g. a ferromagnetic group
- a radioactive group e. g.
- a group comprising 1 H, 14 C, 32 P, 35 S or 125 I or any combination thereof; or to a globulomer flagged by being covalently or by non-covalent high-affinity interaction, preferably covalently linked to a group that facilitates inactivation, sequestration, degradation and/or precipitation, preferably flagged with a group that promotes in vivo degradation, more preferably with ubiquitin, where is particularly preferred if this flagged oligomer is assembled in vivo; or to a globulomer modified by any combination of the above.
- Such labelling and flagging groups and methods for attaching them to proteins are known in the art. Labelling and/or flagging may be performed before, during or after globulomerisation.
- a globulomer derivative is a molecule obtainable from a globulomer by a labelling and/or flagging reaction.
- a ⁇ (X-Y) monomer derivative here refers in particular to an A ⁇ mono- mer that is labelled or flagged as described for the globulomer.
- the antibody of the present invention binds to an A ⁇ (20-42) globulomer with a K 0 in the range of 1x10 ⁇ 6 M to 1x10 ⁇ 12 M.
- the antibody binds to an A ⁇ (20-42) globulomer with high affinity, for instance with a K D of 1x10 ⁇ 7 M or greater affinity, e.g. with a K D of 3x10 "8 M or greater affinity, with a K D of 1 x10 ⁇ 8 M or greater affinity, e.g. with a K D of 3x10 "9 M or greater affinity, with a K D of 1x10 ⁇ 9 M or greater affinity, e.g.
- K D of 3x10 ⁇ 10 M or greater affinity with a K 0 of 1x10 "10 M or greater affinity, e.g. with a K D of 3x10 ⁇ 11 M or greater affinity, or with a K D of 1x10 ⁇ 11 M or greater affinity.
- greater affinity here refers to a degree of interaction where the equilibrium between unbound antibody and unbound globulomer on the one hand and antibody-globulomer complex on the other is further in favour of the antibody-globulomer complex.
- small affinity here refers to a degree of interaction where the equilibrium between unbound antibody and unbound globulomer on the one hand and antibody-globulomer complex on the other is further in favour of the unbound antibody and unbound globulomer.
- greater affinity is synonymous with the term “higher affinity” and term “smaller affinity”is synonymous with the term “lower affinity”.
- the invention relates to an antibody which binds to the A ⁇ (20-42) globulomer with a K D in the range of 1x10 ⁇ 6 M to 1x10 ⁇ 12 M, to the A ⁇ (1-42) globu- lomer with a K D of 10 ⁇ 12 M or smaller affinity, the binding affinity to the A ⁇ (20-42) globulomer being greater than the binding affinity to the A ⁇ (1-42) globulomer.
- the binding affinity of the antibody of the present invention to the A ⁇ (20-42) globulomer is at least 2 times, e. g. at least 3 times or at least 5 times, preferably at least 10 times, e. g. at least 20 times, at least 30 times or at least 50 times, more preferably at least 100 times, e. g. at least 200 times, at least 300 times or at least 500 times, and even more preferably at least 1000 times, e. g. at least 2000 times, at least 3000 times or at least 5000 times, even more preferably at least 10000 times, e. g. at least 20000 times, at least 30000 or at least 50000 times, and most preferably at least 100000 times greater than the binding affinity of the antibody to the A ⁇ (1 -42) globulomer.
- the invention relates to an antibody which binds to the A ⁇ (12-42) globulomer with a K D with a K 0 of 10 ⁇ 12 M or smaller affinity, the binding affinity to the A ⁇ (20-42) globulomer being greater than the binding affinity to the A ⁇ (12-42) globulomer.
- the binding affinity of the antibody of the present invention to the A ⁇ (20- 42) globulomer is at least 2 times, e. g. at least 3 times or at least 5 times, preferably at least 10 times, e. g. at least 20 times, at least 30 times or at least 50 times, more preferably at least 100 times, e. g. at least 200 times, at least 300 times or at least 500 times, and even more preferably at least 1000 times, e. g. at least 2000 times, at least 3000 times or at least 5000 times, even more preferably at least 10000 times, e. g. at least 20000 times, at least 30000 or at least 50000 times, and most preferably at least 100000 times greater than the binding affinity of the antibody to the A ⁇ (12-42) globulomer.
- the antibodies of the present invention bind to at least one A ⁇ globulomer, as defined above, and have a comparatively smaller affinity for at least one non-globulomer form of A ⁇ .
- Antibodies of the present invention having a comparatively smaller affinity for at least one non- globulomer form of A ⁇ than for at least one A ⁇ globulomer include antibodies having a binding affinity to the A ⁇ (20-42) globulomer that is greater than to an A ⁇ (1-42) monomer. Further, it is preferred that, alternatively or additionally, the binding affinity of the antibody to the A ⁇ (20-42) globulomer is greater than to an A ⁇ (1-40) monomer.
- the affinity of the antibody to the A ⁇ (20-42) globulomer is greater than its affinity to both the A ⁇ (1-40) and the A ⁇ (1-42) monomer.
- a ⁇ (X-Y) monomer here refers to the isolated form of the A ⁇ (X-Y) peptide, prefera- bly a form of the A ⁇ (X-Y) peptide which is not engaged in essentially non-covalent interactions with other A ⁇ peptides.
- the A ⁇ (X-Y) monomer is usually provided in the form of an aqueous solution.
- the aqueous monomer solution contains 0.05% to 0.2%, more preferably about 0.1% NH 4 OH.
- the aqueous monomer solution contains 0.05% to 0.2%, more preferably about 0.1% NaOH.
- a ⁇ (1-40) monomer here refers to an A ⁇ (1-40) monomer preparation as described in example 2 herein
- a ⁇ (1-42) monomer here refers to an A ⁇ (1-42) preparation as described in example 2 herein.
- the antibody of the present invention binds to one or, more preferably, both mono- mers with low affinity, most preferably with a K D of 1x10 ⁇ 8 M or smaller affinity, e. g. with a K D of 3x10 "8 M or smaller affinity, with a K D of 1x10 ⁇ 7 M or smaller affinity, e. g. with a K D of 3x1 Or 7 M or smaller affinity, or with a K D of 1x10 ⁇ 6 M or smaller affinity, e. g. with a K D of 3x10 "5 M or smaller affinity, or with a K 0 of 1x10 ⁇ 5 M or smaller affinity.
- the binding affinity of the antibody of the present invention to the A ⁇ (20-42) globulomer is at least 2 times, e. g. at least 3 times or at least 5 times, preferably at least 10 times, e. g. at least 20 times, at least 30 times or at least 50 times, more preferably at least 100 times, e. g. at least 200 times, at least 300 times or at least 500 times, and even more preferably at least 1000 times, e. g. at least 2000 times, at least 3000 times or at least 5000 times, even more preferably at least 10000 times, e. g. at least 20000 times, at least 30000 or at least 50000 times, and most preferably at least 100000 times greater than the binding affinity of the antibody to one or, more preferably, both monomers.
- Antibodies of the present invention having a comparatively smaller affinity for at least one non- globulomer form of A ⁇ than for at least one A ⁇ globulomer further include antibodies having a binding affinity to the A ⁇ (20-42) globulomer that is greater than to A ⁇ (1-42) fibrils. Further, it is preferred that, alternatively or additionally, the binding affinity of the antibody to the A ⁇ (20-42) globulomer is greater than to A ⁇ (1-40) fibrils.
- fibril here refers to a molecular structure that comprises assemblies of non- covalently associated, individual A ⁇ (X-Y) peptides, which show fibrillary structure in the electron microscope, which bind Congo red and then exhibit birefringence under polarized light and whose X-ray diffraction pattern is a cross- ⁇ structure.
- a fibril is a molecular structure obtainable by a process that comprises the self-induced polymeric aggregation of a suitable A ⁇ peptide in the absence of detergents, e. g. in 0.1 M HCI, leading to the formation of aggregates of more than 24, preferably more than 100 units. This process is well known in the art.
- a ⁇ (X-Y) fibrils are used in the form of an aqueous solution.
- the aqueous fibril solution is made by dissolving the A ⁇ peptide in 0.1% NH 4 OH, diluting it 1 : 4 with 20 mM NaH 2 PO 4 , 140 mM NaCI, pH 7.4, followed by readjusting the pH to 7.4, incubating the solution at 37 0 C for 20 h, followed by centrifugation at 10000 g for 10 min and re- suspension in 20 mM NaH 2 PO 4 , 140 mM NaCI, pH 7.4.
- a ⁇ (X-Y) fibril here refers to a fibril consisting essentially of A ⁇ (X-Y) subunits, where it is preferred if on average at least 90% of the subunits are of the A ⁇ (X-Y) type, more preferred if at least 98% of the subunits are of the A ⁇ (X-Y) type, and most preferred if the content of non-A ⁇ (X-Y) peptides is below the detection threshold.
- a ⁇ (1-42) fibril here refers to a A ⁇ (1-42) fibril preparation as described in example 3 herein.
- the antibody of the present invention binds to one or, more preferably, both fibrils with low affinity, most preferably with a K D of IxIO "8 M or smaller affinity, e. g. with a K D of
- 3x10 "8 M or smaller affinity with a K D of 1x10 ⁇ 7 M or smaller affinity, e. g. with a K 0 of 3x10 ⁇ 7 M or smaller affinity, or with a K 0 of 1x10 ⁇ 6 M or smaller affinity, e. g. with a K D of 3x10 ⁇ 5 M or smaller affinity, or with a K D of 1x10 ⁇ 5 M or smaller affinity.
- a ⁇ (20-42) globulomer is at least 2 times, e. g. at least 3 times or at least 5 times, preferably at least 10 times, e. g. at least 20 times, at least 30 times or at least 50 times, more preferably at least 100 times, e. g. at least 200 times, at least 300 times or at least 500 times, and even more preferably at least 1000 times, e. g. at least 2000 times, at least 3000 times or at least 5000 times, even more preferably at least 10000 times, e. g. at least 20000 times, at least 30000 or at least 50000 times, and most preferably at least 100000 times greater than the binding affinity of the antibody to one or, more preferably, both fibrils.
- the invention relates to antibodies having a binding affinity to the A ⁇ (20-42) globulomer which is greater than its binding affinity to both A ⁇ (1-40) and A ⁇ (1 -42) fibrils.
- the present invention relates to antibodies having a comparatively smaller affinity for both the monomeric and fibrillary forms of A ⁇ than for at least one A ⁇ globulomer, in particular A ⁇ (20-42) globulomer.
- a ⁇ globulomer in particular A ⁇ (20-42) globulomer.
- Antibodies of the present invention further include antibodies having a binding affinity to the A ⁇ (20-42) globulomer that is greater than to a cross-linked A ⁇ (1-42) globulomer, in particular to a glutardialdehyde cross-linked A ⁇ (1-42) globulomer, such as that described in example 1b herein.
- the binding affinity of the antibody to A ⁇ (20-42) globulomer is at least 2 times, e. g. at least 3 times or at least 5 times, preferably at least 10 times, e. g. at least 20 times, at least 30 times or at least 50 times, more preferably at least 100 times, e. g. at least 200 times, at least 300 times or at least 500 times, and even more preferably at least 1000 times, e. g. at least 2000 times, at least 3000 times or at least 5000 times, even more preferably at least 10000 times, e. g. at least 20000 times, at least 30000 or at least 50000 times, and most preferably at least 100000 times greater than the binding affinity of the antibody to cross-linked A ⁇ (1-42) globulomer.
- the antibodies of the present invention are preferably isolated, in particular monoclonal and more particularly recombinant.
- the present invention also relates to a monoclonal antibody (5F7) that is obtainable from a hybridoma designated by American Type Culture Collection deposit number PTA-7241.
- the present invention also relates to a monoclonal antibody (10F11 ) that is obtainable from a hybridoma designated by American Type Culture Collection deposit number PTA-7239.
- the present invention also relates to a monoclonal antibody (7C6) that is obtainable from a hybridoma designated by American Type Culture Collection deposit number PTA-7240.
- the present invention also relates to a monoclonal antibody (4B7) that is obtainable from a hybridoma designated by American Type Culture Collection deposit number PTA-7242.
- the present invention also relates to a monoclonal antibody (6A2) that is obtainable from a hybridoma designated by American Type Culture Collection deposit number PTA-7409.
- the present invention also relates to a monoclonal antibody (2F2) that is obtainable from a hybridoma designated by American Type Culture Collection deposit number PTA-7408.
- the present invention also relates to a monoclonal antibody (4D10) that is obtainable from a hybridoma designated by American Type Culture Collection deposit number PTA-7405.
- the present invention also relates to a monoclonal antibody (7E5) that is obtainable from a hybridoma designated by American Type Culture Collection deposit number PTA-7809.
- the present invention also relates to a monoclonal antibody (10C1 ) that is obtainable from a hybridoma designated by American Type Culture Collection deposit number PTA-7810.
- the present invention also relates to a monoclonal antibody (3B10) that is obtainable from a hybridoma designated by American Type Culture Collection deposit number PTA-7851.
- antibodies of the present invention are characterized by having a binding affinity to an A ⁇ (20-42) globulomer that is greater than the binding affinity of the antibody to an A ⁇ (1-42) globulomer.
- the present invention also relates to antibodies having a similar binding profile to that of any one of said monoclonal antibodies, 5F7, 10F11 , 7C6, 4B7, 6A2, 2F2, 4D10, 7E5, 10C1 and 3B10.
- Antibodies having a similar binding profile to that of any one of said monoclonal antibodies should be understood as not being limited to antibodies having a binding affinity to an A ⁇ (20-42) globulomer that is greater than the binding affinity of the antibody to an A ⁇ (1-42) globulomer.
- Antibodies having a binding profile similar to that of any one of said monoclonal antibodies, 5F7, 10F11 , 7C6, 4B7, 6A2, 2F2, 4D10, 7E5, 10C1 and 3B10 include antibodies which bind to the same epitope as monoclonal antibody 5F7, 10F11 , 7C6, 4B7, 6A2, 2F2, 4D10, 7E5, 10C1 and 3B10.
- All monoclonal antibodies from the group consisting of 5F7, 10F11 , 7C6, 4B7, 6A2, 2F2, 4D10, 7E5, 10C1 and 3B10 bind to an epitope contained within the 20 and 42 A ⁇ sequence range, in particular within the 20-30 A ⁇ sequence range.
- said epitope is believed to be a structural, non-linear epitope in between subunits in the region of amino acids 20 and 42, in particular in the region of amino acids 20 and 30.
- the present invention also relates to antibodies which are capable of competing with at least one, preferably all, antibodies selected from the group consisting of 5F7, 10F11 , 7C6, 4B7, 6A2, 2F2, 4D10, 7E5, 10C1 and 3B10.
- serving antibodies refers to any number of antibodies targeting the same molecular or stably but non-covalently linked supermolecular entity, preferably the same molecule, wherein at least one is capable of specifically reducing the measurable binding of an- other, preferably by sterically hampering the other's access to its target epitope or by inducing and/or stabilizing a conformation in the target entity that reduces the target's affinity for the other antibody, more preferably by directly blocking access to the other's target epitope by binding to an epitope in sufficiently close vicinity of the former, overlapping with the former or identical to the former, most preferably overlapping or identical, in particular identical.
- Two epi- topes are herein said to be “overlapping” if they share part of their chemical structures, preferably their amino acid sequences, and to be “identical”, if their chemical structures, preferably their amino acid sequences, are identical.
- the present invention also relates to antibodies whose target epitopes are overlapping with, preferably identical to, the target epitope of at least one of the antibodies selected from the group consisting of 5F7, 10F11 , 7C6, 4B7, 6A2, 2F2, 4D10, 7E5, 10C1 and 3B10.
- Antibodies having a similar binding profile to that of any one of said monoclonal antibodies, 5F7, 10F11 , 7C6, 4B7, 6A2, 2F2, 4D10, 7E5, 10C1 and 3B10 thus further include antibodies which comprise at least a portion of the antigen-binding moiety of any one of said monoclonal antibodies, 5F7, 10F11 , 7C6, 4B7, 6A2, 2F2, 4D10, 7E5, 10C1 and 3B10.
- said portion comprises at least one complementary determining region (CDR) of any one of said monoclonal antibodies, 5F7, 10F11 , 7C6, 4B7, 6A2, 2F2, 4D10, 7E5, 10C1 and 3B10.
- the present invention relates to antibodies comprising the amino acid sequence of the heavy chain CDR3 and/or the amino acid sequence of the light chain CDR3 of monoclonal antibody 5F7, 10F11 , 7C6, 4B7, 6A2, 2F2, 4D10, 7E5, 10C1 or 3B10.
- antibodies include those which also comprise the amino acid sequence of the heavy chain CDR2 and/or the amino acid sequence of the light chain CDR2 of monoclonal antibody 5F7, 10F11 , 7C6, 4B7, 6A2, 2F2, 4D10, 7E5, 10C1 or 3B10, respectively.
- such antibodies include those which also comprise the amino acid sequence of the heavy chain CDR1 and/or the amino acid sequence of the light chain CDR1 of monoclonal antibody 5F7, 10F11 , 7C6, 4B7, 6A2, 2F2, 4D10, 7E5, 10C1 or 3B10, respectively.
- the present invention thus relates to antibodies comprising a heavy chain wherein the CDR3, CDR2 and/or CDR1 domain comprises the amino acid sequence of the heavy chain CDR3, CDR2 and/or CDR1 of monoclonal antibody 5F7, 10F11 , 7C6, 4B7, 6A2, 2F2, 4D10, 7E5, 10C1 or 3B10.
- the present invention thus relates to antibodies comprising a light chain wherein the CDR3, CDR2 and/or CDR1 domain comprises the amino acid sequence of the light chain CDR3, CDR2 and/or CDR1 , respectively, of monoclonal antibody 5F7, 10F11 , 7C6, 4B7, 6A2, 2F2, 4D10, 7E5, 10C1 or 3B10.
- the antibody comprises at least one CDR comprising an amino acid sequence selected from the group consisting of: amino acid residues 31-35 of SEQ ID NO:3, amino acid residues 50-66 of SEQ ID NO:3, amino acid residues 99-109 of SEQ ID NO:3, amino acid residues 24-39 of SEQ ID NO:4, amino acid residues 55-61 of SEQ ID NO:4, amino acid residues 94-102 of SEQ ID NO:4, amino acid residues 31-35 of SEQ ID NO:7, amino acid residues 50- 66 of SEQ ID NO:7, amino acid residues 97-109 of SEQ ID NO:7, amino acid residues 24-39 of SEQ ID NO:8, amino acid residues 55-61 of SEQ ID NO:8, amino acid residues 94-102 of SEQ ID NO:8, amino acid residues 31-35 of SEQ ID NO:11 , amino acid residues 50-65 of SEQ ID NO:11 , amino acid residues 98-107 of SEQ ID NO:11 , amino acid residues 24-39 of
- the antibody comprises at least 3 CDRs selected from the group consisting of the sequences disclosed above. More preferably the 3 CDRs selected are from sets of variable domain CDRs selected from the group consisting of:
- the antibody of the invention comprises at least two variable domain CDR sets. More preferably, the two variable domain CDR sets are selected from the group consisting of: VH 5F7 CDR Set & VL 5F7 CDR Set; VH 10F11 CDR Set & VL 10F11 CDR Set; VH 7C6 CDR Set & VL 7C6 CDR Set; VH 4B7 CDR Set & VL 4B7 CDR Set; VH 2F2 CDR Set & VL 2F2 CDR Set; VH 6A2 CDR Set & VL 6A2 CDR Set; VH 4D10 CDR Set & VL 4D10 CDR Set; VH 7E5 CDR Set & VL 7E5 CDR Set; and VH 10C1 CDR Set & VL 10C1 CDR Set.
- the antibody disclosed above further comprises a human acceptor framework.
- the antibody is a CDR grafted antibody.
- the CDR grafted antibody comprises one or more of the CDRs disclosed above.
- the CDR grafted antibody comprises a human acceptor framework.
- the antibody is a humanized antibody.
- the humanized antibody comprises one or more of the CDRs disclosed above. More preferably the humanized antibody comprises three or more of the CDRs disclosed above. Most preferably the human- ized antibody comprises six CDRs disclosed above.
- the CDRs are incorporated into a human antibody variable domain of a human acceptor framework.
- the human antibody variable domain is a consensus human variable domain.
- the human acceptor framework comprises at least one Framework Region amino acid substitution at a key residue, wherein the key residue is selected from the group consisting of a residue adjacent to a CDR; a glycosylation site residue; a rare residue; a residue capable of interacting with A ⁇ (20-42) globulomer; a residue capable of interacting with a CDR; a canonical residue; a contact residue between heavy chain variable region and light chain variable region; a residue within a Vernier zone; and a residue in a region that overlaps between a Chothia-defined variable heavy chain CDR1 and a Kabat-defined first heavy chain framework.
- the human acceptor framework human acceptor framework comprises at least one Framework Region amino acid substitution, wherein the amino acid sequence of the framework is at least 65% identical to the sequence of said human acceptor framework and comprises at least 70 amino acid residues identical to said human acceptor framework.
- the present invention relates to antibodies comprising both the heavy and light chain as defined above.
- the antibody comprises at least one variable domain having an amino acid sequence selected from the group consisting of: SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:11 , SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:31 , SEQ ID NO:32, SEQ ID NO:35, SEQ ID NO:36, and SEQ ID NO:38.
- the antibody comprises two variable domains, wherein said two variable domains have amino acid sequences selected from the group consisting of: SEQ ID NO:3 & SEQ ID NO:4, SEQ ID NO:7 & SEQ ID NO:8 & SEQ ID NO:11 & SEQ ID NO:12, SEQ ID NO:15 & SEQ ID NO:16, SEQ ID NO: 19 & SEQ ID NO:20, SEQ ID NO:23 & SEQ ID NO:24, SEQ ID NO:27 & SEQ ID NO:28, SEQ ID NO:31 & SEQ ID NO:32, and SEQ ID NO:35 & SEQ ID NO:36.
- the antibodies of the present invention comprise a heavy chain constant region selected from the group consisting of IgGI , lgG2, lgG3, lgG4, IgM, IgA, IgD, IgE and human IgGI Ala234 Ala235 mutant constant regions.
- the antibodies comprise a human constant region. More preferably, the antibodies comprise an amino acid sequence selected from the group consisting of SEQ ID NOs:39-42. Antibodies comprising an IgGI heavy chain constant region are preferred.
- the antibody is glycosylated.
- the glycosylation pattern is a human glycosylation pattern or a glycosylation pattern produced by any one of the eukaryotic cells disclosed herein, in particular CHO cells.
- the present invention also relates to an antigen-binding moiety of an antibody of the present invention.
- antigen-binding moieties include, but are not limited to, Fab fragments, F(ab') 2 fragments and single chain Fv fragments of the antibody. Further antigen-binding moieties are Fab' fragments, Fv fragments, and disulfide linked Fv fragments.
- the invention also provides an isolated nucleic acid encoding any one of the antibodies disclosed herein.
- a further embodiment provides a vector comprising the isolated nucleic acid disclosed herein.
- Said vector may in particular be selected from the group consisting of pcDNA; pTT (Durocher et al., Nucleic Acids Research 2002, VoI 30, No.2); pTT3 (pTT with additional multiple cloning site; pEFBOS (Mizushima, S. and Nagata, S., (1990) Nucleic Acids Research VoI 18, No. 17); pBV; pJV; and pBJ.
- a host cell is transformed with the vector disclosed herein.
- the host cell is a prokaryotic cell. More preferably the host cell is E.coli.
- the host cell is a eukaryotic cell.
- the eukaryotic cell is selected from the group consisting of a protist cell, an animal cell (e.g. a mammalian cell, an avian cell, and an insect cell), a plant cell and a fungal cell. More preferably the host cell is a mammalian cell including, but not limited to, CHO and COS; or a fungal cell, eg., a yeast cell, such as Saccharomyces cere- visiae; or an insect cell such as Sf9.
- Another aspect of the invention provides a method of producing an antibody of the invention, comprising culturing any one of the host cells or a hybridoma disclosed herein in a culture medium under conditions suitable to produce the antibody.
- Another embodiment provides an antibody that is obtainable by the method disclosed herein.
- Antibodies of the present invention can be obtained in a manner known per se.
- B lymphocytes which, in totality, contain an antibody repertoire composed of hundreds of billions of different antibody specificities are a part of the mammalian immune system.
- a normal immune response to a particular antigen means selection of one or more antibodies of said repertoire which specifically bind to said antigen, and the success of an immune response is based at least partially on the ability of said antibodies to specifically recognize (and ultimately to eliminate) the stimulating antigen and to ignore other molecules in the environment of said antibodies.
- Standardized hybridoma technology now allows the production of antibodies with a single specificity for an antigen of interest. More recently, recombinant antibody techniques such as in-vitro screening of antibody libraries have been developed. These techniques likewise allow antibodies having a single specificity for an antigen of interest to be produced.
- the antigen of interest may be allowed to act on the antibody repertoire either in vivo or in vitro.
- the antigen is allowed to act on the repertoire by immunizing an animal in vivo with said antigen.
- This in-vivo approach may furthermore comprise establishing from the lymphocytes of an animal a number of hybridomas and selecting a particular hybridoma which secretes an antibody specifically binding to said antigen.
- the animal to be immunized may be, for example, a mouse, rat, rabbit, chicken, camelid or sheep or may be a transgenic version of any of the animals mentioned above, for example a transgenic mouse with human immunoglobulin genes, which produces human antibodies after an antigenic stimulus.
- mice with severe combined immunodeficiency which have been reconstituted with human peripheral mononuclear blood cells (chimeric hu-PBMC SCID mice) or with lymphoid cells or precursors thereof, as well as mice which have been treated with a lethal total body irradiation, then protected against radia- tion with bone marrow cells from a mouse with severe combined immunodeficiency (SCID) and subsequently transplanted with functional human lymphocytes (the "Trimera” system).
- SCID severe combined immunodeficiency
- Another type of an animal to be immunized is an animal (e.g.
- polyclonal or monoclonal antibodies produced by this method are characterized and selected by using known screening methods which include, but are not limited to, ELISA and dot blot techniques.
- the antigen is allowed to act on the antibody repertoire in vitro by screening a recombinant antibody library with said antigen.
- the recombinant antibody library may be expressed, for example, on the surface of bacteriophages or on the surface of yeast cells or on the surface of bacterial cells.
- the recombinant antibody library is an scFv library or an Fab library, for example.
- antibody libraries are expressed as RNA-protein fusions.
- the antigen may be allowed to act on the antibody repertoire by immunizing an animal in vivo with said antigen and then screening in vitro with said antigen a recombinant antibody library prepared from lymphoid cells of said animal or a single domain antibody library (e.g. containing heavy and/or light chains).
- the antigen is allowed to act on the antibody repertoire by immunizing an animal in vivo with said antigen and then subjecting a recombinant antibody library or single domain library produced from lymphoid cells of said animal to affinity maturation.
- the antigen is allowed to act on the antibody repertoire by immunizing an animal in vivo with said antigen, then selecting individual antibody-producing cells secreting an antibody of interest and obtaining from said selected cells cDNAs for the variable region of the heavy and light chains (e.g. by means of PCR) and expressing said variable regions of the heavy and light chains in mammalian host cells in vitro (this being referred to as selected lymphocyte antibody method or SLAM), thereby being able to further select and manipulate the selected antibody gene sequences.
- monoclonal antibodies may be selected by expression cloning by expressing the antibody genes for the heavy and light chains in mammalian cells and selecting those mammalian cells which secrete an antibody having the desired binding affinity.
- the present invention provides defined antigens for screening and counter screening. Thus it is possible, according to the invention, to select those polyclonal and monoclonal antibodies which bind to an A ⁇ (20-42) globulomer with the binding affinities as defined above.
- the methods of the invention for producing antibodies can be used to produce various types of antibodies. These include monoclonal, in particular recombinant antibodies, especially essen- tially human antibodies, chimeric antibodies, humanized antibodies and CDR graft antibodies, and also antigen-binding moieties thereof.
- the present invention further relates to a hybridoma that is capable of producing (secreting) a monoclonal antibody of the present invention.
- Hybridomas of the present invention include those designated by an American Type Culture Collection deposit number selected from the group consisting of PTA-7241 , PTA-7239, PTA-7240, PTA-7242, PTA-7408, PTA-7409, PTA- 7405, PTA-7809, PTA-7810 and PTA-7851.
- the antibodies of the present invention may also be reactive with, i.e. bind to, A ⁇ forms other than the A ⁇ globulomers described herein. These antigens may or may not be oligomeric or globulomeric. Thus, the antigens to which the antibodies of the present invention bind include any A ⁇ form that comprises the globulomer epitope with which the antibodies of the present invention are reactive.
- Such A ⁇ forms include truncated and non-truncated A ⁇ (X- Y) forms (with X and Y being defined as above), such as A ⁇ (20-42), A ⁇ (20-40), A ⁇ (12-42), A ⁇ (12-40), A ⁇ (1-42), and A ⁇ (1-40) forms, provided that said forms comprise the globulomer epitope.
- the present invention also relates to a composition
- a composition comprising an antibody of the invention or an antigen-binding moiety thereof, as defined above.
- said composition is a pharmaceutical composition which comprises the antibody of the invention or the antigen-binding moiety and a pharmaceutical acceptable carrier.
- the antibody of the invention or the antigen-binding moiety as defined above is preferably capable of neutralizing, both in vitro and in vivo, the activity of A ⁇ globulomer or a derivative thereof to which it binds.
- Said antibody or antigen-binding moiety may therefore be used for inhibiting the activity of said globulomer or derivative thereof, for example in a preparation con- taining said globulomer or derivative thereof or in human individuals or other mammals in which said globulomer or derivative thereof is present.
- the invention relates to a method of inhibiting the activity of said globulomer or derivative thereof, which method comprises allowing an antibody of the inven- tion or an antigen-binding moiety thereof to act on a globulomer or derivative thereof so as to inhibit the activity of said globulomer or derivative thereof.
- Said activity may be inhibited in vitro, for example.
- the antibody of the invention or the antigen-binding moiety may be added to a preparation such as a sample derived from a subject or a cell culture which contains or is suspected to contain said globulomer or derivative thereof, in order to inhibit the activity of said globulomer or derivative thereof in said sample.
- the activity of the globulomer or derivative thereof may be inhibited in an individual in vivo.
- the present invention further relates to the use of an antibody or an antigen-binding moi- ety as defined above for preparing a pharmaceutical composition for treating or preventing an amyloidosis, in particular an amyloidosis selected from the group consisting of Alzheimer's disease and the amyloidosis of Down's syndrome.
- an antibody or an antigen-binding moi- ety as defined above for preparing a pharmaceutical composition for treating or preventing an amyloidosis, in particular an amyloidosis selected from the group consisting of Alzheimer's disease and the amyloidosis of Down's syndrome.
- One aspect of said use of the invention is therefore a method of treating or preventing an amyloidosis, in particular Alzheimer's disease or the amyloidosis of Down's syndrome, in a subject in need thereof, which comprises adminis- tering an antibody or an antigen-binding moiety as defined above to the subject.
- said antibody or antigen-binding moiety for treating and especially preventing the amyloidosis, in particular Alzheimer's disease or the amyloidosis of Down's syndrome is in particular for passive immunization. Accordingly, in the method of treating or preventing an amyloidosis, in particular Alzheimer's disease or the amyloidosis of Down's syndrome, in a subject in need thereof one purpose of administering the antibody or antigen-binding moiety to the subject is passively immunizing the subject against the amyloidosis, in particular Alzheimer's disease or the amyloidosis of Down's syndrome.
- the antibody of the invention or the antigen-binding moiety as defined above is preferably ca- pable of detecting, both in vitro and in vivo, an A ⁇ globulomer or derivative thereof to which it binds.
- Said antibody or the antigen-binding moiety may therefore be used for detecting said globulomer or derivative thereof, for example in a preparation containing said globulomer or derivative thereof or in human individuals or other mammals in which said globulomer or derivatives thereof is present.
- the invention relates to a method of detecting said globulomer or derivative thereof, which method comprises allowing an antibody of the invention or an antigen-binding moiety thereof to act on a globulomer or derivative thereof so as to bind to said globulomer or derivative thereof (and thereby preferably forming a complex comprising the antibody or antigen-binding moiety thereof and the globulomer or derivative thereof).
- Said globulomer may be detected in vitro, for example.
- the antibody of the invention or the antigen-binding moiety may be added to a preparation, for instance a sample derived from a subject or a cell culture which contains or is suspected to contain said globulomer or derivative thereof, in order to detect said globulomer or derivative thereof in said preparation.
- a preparation for instance a sample derived from a subject or a cell culture which contains or is suspected to contain said globulomer or derivative thereof, in order to detect said globulomer or derivative thereof in said preparation.
- the globulomer or derivative thereof may be detected in an individual in vivo.
- the present invention further relates to the use of an antibody or an antigen-binding moiety as defined above for preparing a composition for diagnosing an amyloidosis, in particular Alzheimer's disease or the amyloidosis of Down's syndrom.
- One aspect of said use of the in- vention is a method of diagnosing an amyloidosis, in particular Alzheimer's disease or the amyloidosis of Down's syndrom, in a subject suspect of having the amyloidosis, in particular Alzheimer's disease or the amyloidosis of Down's syndrom, which comprises administering to the subject an antibody or an antigen-binding moiety as defined above and detecting the formation of a complex comprising the antibody or the antigen-binding moiety with the antigen, the presence of the complex indicating the amyloidosis, in particular Alzheimer's disease or the amyloidosis of Down's syndrom, in the subject.
- a second aspect of said use of the invention is a method of diagnosing an amyloidosis, in particular Alzheimer's disease or the amyloidosis of Down's syndrom, in a subject suspect of having the amyloidosis, in particular Alzheimer's disease or the amyloidosis of Down's syndrom, which comprises providing a sample from the subject, contacting the sample with an antibody or an antigen-binding moiety as defined above and detecting the formation of a complex comprising the antibody or the antigen-binding moiety with the antigen, the presence of the complex indicating the amyloidosis, in particular Alzheimer's disease or the amyloidosis of Down's syndrom, in the subject.
- the binding affinities of the antibodies of the invention may be evaluated by using standardized in-vitro immunoassays such as ELISA, dot blot or BIAcore analyses (Pharmacia Biosen- sor AB, Uppsala, Sweden and Piscataway, NJ).
- ELISA ELISA
- dot blot BIAcore analyses
- Johnsson, B., et al. (1995) J. MoI. Recognit. 8:125-131 and Johnsson, B., et al. (1991 ) Anal. Biochem. 198:268-277.
- the affinities defined herein refer to the values obtained by performing a dot blot as described in example 8 and evaluating it by densitometry.
- determining the binding affinity by dot blot comprises the following: a certain amount of the antigen (e.g.
- the relative affinity of two different antibodies to one target, or of one antibody to two different targets is here defined as the relation of the respective amounts of target-bound antibody observed with the two antibody-target combinations under otherwise identical dot blot conditions.
- the dot blot approach will determine an antibody's affinity to a given target in the latter's natural conformation; unlike the ELISA approach, the dot blot approach does not suffer from differences in the affinities between different targets and the matrix, thereby allowing for more precise comparisons between different targets.
- Kd is intended to refer to the dissociation constant of a particular antibody-antigen interaction as is known in the art.
- the antibodies of the present invention are preferably isolated antibodies.
- An "isolated antibody” means an antibody having the binding affinities as described above and which is essen- tially free of other antibodies having different binding affinities.
- the term “essentially free” here refers to an antibody preparation in which at least 95% of the antibodies, preferably at least 98% of the antibodies and more preferably at least 99% of the antibodies have the desired binding affinity.
- an isolated antibody may be substantially free of other cellular material and/or chemicals.
- the isolated antibodies of the present invention include monoclonal antibodies.
- a "monoclonal antibody” as used herein is intended to refer to a preparation of antibody molecules, antibodies which share a common heavy chain and common light chain amino acid sequence, in contrast with “polyclonal” antibody preparations which contain a mixture of antibodies of different amino acid sequence.
- Monoclonal antibodies can be generated by several novel technologies like phage, bacteria, yeast or ribosomal display, as well as by classical methods exemplified by hybridoma-derived antibodies (e.g., an antibody secreted by a hybridoma prepared by hybri- doma technology, such as the standard Kohler and Milstein hybridoma methodology ((1975) Nature 256:495-497).
- a non-hybridoma-derived antibody with uniform sequence is still referred to as a monoclonal antibody herein although it may have been obtained by non- classical methodologies, and the term "monoclonal" is not restricted to hybridoma-derived antibodies but used to refer to all antibodies derived from one nucleic acid clone.
- the monoclonal antibodies of the present invention include recombinant antibodies.
- the term "recombinant” herein refers to any artificial combination of two otherwise separated segments of sequence, e.g., by chemical synthesis or by the manipulation of isolated segments of nucleic acids by genetic engineering techniques.
- the term “recombinant antibody” refers to antibodies which are produced, expressed, generated or isolated by recombinant means, such as antibodies which are expressed using a recombinant expression vector trans- fected into a host cell; antibodies isolated from a recombinant combinatorial antibody library; antibodies isolated from an animal (e.g. a mouse) which is transgenic due to human immunoglobulin genes (see, for example, Taylor, L.D., et al.
- Recombinant antibodies include, for example, chi- meric, CDR graft and humanized antibodies.
- expression of a conventional hybridoma-derived monoclonal antibody in a heterologous system will require the generation of a recombinant antibody even if the amino acid sequence of the resulting antibody protein is not changed or intended to be changed.
- the antibody is a humanized antibody.
- the antibody may comprise an amino acid sequence derived entirely from a single species, such as a human antibody or a mouse antibody.
- the antibody may be a chimeric antibody or a CDR graft antibody or another form of a humanized antibody.
- antibody is intended to refer to immunoglobulin molecules consisting of 4 polypeptide chains, two heavy (H) chains and two light (L) chains.
- the chains are usually linked to one another via disulfide bonds.
- Each heavy chain is composed of a variable region of said heavy chain (abbreviated here as HCVR or VH) and a constant region of said heavy chain.
- the heavy chain constant region consists of three domains CH1 , CH2 and CH3.
- Each light chain is composed of a variable region of said light chain (abbreviated here as LCVR or VL) and a constant region of said light chain.
- the light chain constant region consists of a CL domain.
- VH and VL regions may be further divided into hypervariable regions referred to as complementarity-determining regions (CDRs) and interspersed with conserved regions referred to as framework regions (FR).
- CDRs complementarity-determining regions
- FR framework regions
- Each VH and VL region thus consists of three CDRs and four FRs which are arranged from the N terminus to the C terminus in the following order: FR1 , CDR1 , FR2, CDR2, FR3, CDR3, FR4. This structure is well known to those skilled in the art.
- an antibody refers to one or more fragments of an antibody of the invention, said fragment(s) still having the binding affinities as defined above. Fragments of a complete antibody have been shown to be able to carry out the antigen-binding function of an antibody.
- binding fragments include (i) an Fab fragment, i.e. a monovalent fragment composed of the VL, VH, CL and CH1 domains; (ii) an F(ab') 2 fragment, i.e.
- a bivalent fragment comprising two Fab fragments linked to one another in the hinge region via a disulfide bridge; (iii) an Fd fragment composed of the VH and CH1 domains; (iv) an Fv fragment composed of the FL and VH domains of a single arm of an antibody; (v) a dAb frag- ment (Ward ed a/., (1989) Nature 341 :544-546) consisting of a VH domain or of VH, CH1 , CH2, DH3, or VH, CH2, CH3; and (vi) an isolated complementarity-determining region (CDR).
- CDR complementarity-determining region
- the two domains of the Fv fragment namely VL and VH
- VL and VH are encoded by separate genes
- they may further be linked to one another using a synthetic linker, e.g. a poly-G 4 S amino acid sequence, and recombinant methods, making it possible to prepare them as a sin- gle protein chain in which the VL and VH regions combine in order to form monovalent mole- cules (known as single chain Fv (ScFv); see, for example, Bird et al. (1988) Science 242:423- 426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883).
- a synthetic linker e.g. a poly-G 4 S amino acid sequence
- an antibody is also intended to comprise such single chain antibodies.
- Other forms of single chain antibodies such as “diabodies” are likewise included here.
- Diabodies are bivalent, bispecific antibodies in which VH and VL domains are expressed on a single polypeptide chain, but using a linker which is too short for the two domains being able to combine on the same chain, thereby forcing said domains to pair with complementary domains of a different chain and to form two antigen-binding sites (see, for example, Holliger, P., et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, R.J., et al. (1994) Structure 2:1121-1123).
- An immunoglobulin constant domain refers to a heavy or light chain constant domain. Human IgG heavy chain and light chain constant domain amino acid sequences are known in the art and are represented in Table 1.
- Ig gamma-1 SEQ ID NO: 39 ASTKGPSVFFLAPSSKSTSGGTAALGCLVKDY constant region FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
- Ig gamma-1 SEQ ID NO: 40 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY constant region FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS mutant LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK
- an antibody of the present invention or antigen-binding moiety thereof may be part of a larger immunoadhesion molecule formed by covalent or noncovalent association of said antibody or antibody moiety with one or more further proteins or peptides.
- immunoadhesion molecules are the use of the streptavidin core region in order to prepare a tetrameric scFv molecule (Kipriyanov, S. M., et al. (1995) Human Antibodies and Hybri- domas 6:93-101 ) and the use of a cystein residue, a marker peptide and a C-terminal polyhis- tidinyl, e. g.
- human antibody refers to antibodies whose variable and constant regions correspond to or are derived from immunoglobulin sequences of the human germ line, as described, for example, by Kabat et al. (see Kabat, et al. (1991 ) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242).
- the human antibodies of the invention may contain amino acid residues not encoded by human germ line immunoglobulin sequences (for example mutations which have been introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo), for example in the CDRs, and in particular in CDR3.
- Recombinant human antibodies of the invention have variable regions and may also contain constant regions derived from immunoglobulin sequences of the human germ line (see Kabat, E. A., et al. (1991 ) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242).
- recombinant human antibodies are subjected to in-vitro mutagenesis (or to a somatic in-vivo mutagenesis, if an animal is used which is transgenic due to human Ig sequences) so that the amino acid sequences of the VH and VL regions of the recombinant antibodies are sequences which although related to or derived from VH and VL sequences of the human germ line, do not naturally exist in vivo within the human antibody germ line repertoire.
- recombinant antibodies of this kind are the result of selective mutagenesis or back mutation or of both.
- mutagenesis leads to an affinity to the target which is greater, and/or an affinity to non-target structures which is smaller than that of the parent antibody.
- chimeric antibody refers to antibodies which contain sequences for the variable region of the heavy and light chains from one species and constant region sequences from another species, such as antibodies having murine heavy and light chain variable regions linked to human constant regions.
- CDR-grafted antibody refers to antibodies which comprise heavy and light chain variable region sequences from one species but in which the sequences of one or more of the CDR regions of VH and/or VL are replaced with CDR sequences of another species, such as antibodies having murine heavy and light chain variable regions in which one or more of the murine CDRs (e.g., CDR3) has been replaced with human CDR sequences.
- humanized antibody refers to antibodies which contain sequences of the variable region of heavy and light chains from a nonhuman species (e.g. mouse, rat, rabbit, chicken, camelid, sheep or goat) but in which at least one part of the VH and/or VL sequence has been altered in order to be more "human-like", i.e.
- a humanized antibody is a CDR graft antibody in which human CDR sequences have been inserted into nonhuman VH and VL sequences to replace the corresponding nonhuman CDR sequences.
- Kabat numbering “Kabat definitions” and “Kabat labeling” are used interchangea- bly herein. These terms, which are recognized in the art, refer to a system of numbering amino acid residues which are more variable (i.e. hypervariable) than other amino acid residues in the heavy and light chain variable regions of an antibody, or an antigen binding portion thereof (Kabat er a/. (1971 ) ⁇ nn. NY Acad, ScL 190:382-391 and Kabat, E. A., et al. (1991 ) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242).
- the hypervariable region ranges from amino acid positions 31 to 35 for CDR1 , amino acid positions 50 to 65 for CDR2, and amino acid positions 95 to 102 for CDR3.
- the hypervariable region ranges from amino acid positions 24 to 34 for CDR1 , amino acid positions 50 to 56 for CDR2, and amino acid positions 89 to 97 for CDR3.
- the terms “acceptor” and “acceptor antibody” refer to the antibody or nucleic acid sequence providing or encoding at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or 100% of the amino acid sequences of one or more of the framework regions.
- the term “acceptor” refers to the antibody amino acid or nucleic acid se- quence providing or encoding the constant region(s).
- the term “acceptor” refers to the antibody amino acid or nucleic acid sequence providing or encoding one or more of the framework regions and the constant region(s).
- the term "acceptor” refers to a human antibody amino acid or nucleic acid sequence that provides or encodes at least 80%, preferably, at least 85%, at least 90%, at least 95%, at least 98%, or 100% of the amino acid sequences of one or more of the framework regions.
- an acceptor may contain at least 1 , at least 2, at least 3, least 4, at least 5, or at least 10 amino acid residues not occuring at one or more specific positions of a human antibody.
- acceptor framework region and/or acceptor constant region(s) may be, e.g., derived or obtained from a germline antibody gene, a mature antibody gene, a functional anti- body (e.g., antibodies well-known in the art, antibodies in development, or antibodies commercially available).
- CDR refers to the complementarity determining region within antibody variable sequences. There are three CDRs in each of the variable regions of the heavy chain and of the light chain, which are designated CDR1 , CDR2 and CDR3, for each of the variable regions.
- CDR set refers to a group of three CDRs that occur in a single variable region capable of binding the antigen. The exact boundaries of these CDRs have been defined differently according to different systems. The system described by Kabat (Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md.
- CDR boundary definitions may not strictly follow one of the above systems, but will nonetheless overlap with the Kabat CDRs, although they may be shortened or lengthened in light of prediction or experimental findings that particular residues or groups of residues or even entire CDRs do not significantly impact antigen binding.
- the methods used herein may utilize CDRs defined according to any of these systems, although preferred embodiments use Kabat or Chothia defined CDRs.
- canonical residue refers to a residue in a CDR or framework that defines a particular canonical CDR structure as defined by Chothia et al. (J. MoI. Biol. 196:901- 907 (1987); Chothia et al., J. MoI. Biol. 227:799 (1992), both are incorporated herein by reference). According to Chothia et al., critical portions of the CDRs of many antibodies have nearly identical peptide backbone confirmations despite great diversity at the level of amino acid sequence. Each canonical structure specifies primarily a set of peptide backbone torsion angles for a contiguous segment of amino acid residues forming a loop.
- the terms "donor” and “donor antibody” refer to an antibody providing one or more CDRs.
- the donor antibody is an antibody from a species different from the antibody from which the framework regions are obtained or derived.
- the term “donor antibody” refers to a non-human antibody pro- viding one or more CDRs.
- framework or “framework sequence” refers to the remaining sequences of a variable region minus the CDRs. Because the exact definition of a CDR sequence can be determined using different systems, the meaning of a framework sequence is subject to correspondingly different interpretations.
- the six CDRs also divide the framework regions on the light chain and the heavy chain into four sub-regions (FR1 , FR2, FR3 and FR4) on each chain, in which CDR1 is positioned between FR1 and FR2, CDR2 between FR2 and FR3, and CDR3 between FR3 and FR4.
- a framework region represents the combined FR's within the variable region of a single, naturally occurring immunoglobulin chain.
- a FR represents one of the four sub- regions, and FRs represents two or more of the four sub- regions constituting a framework region.
- germline antibody gene or “gene fragment” refers to an immunoglobulin sequence encoded by non- lymphoid cells that have not undergone the maturation process that leads to genetic rearrangement and mutation for expression of a particular immu- noglobulin.
- a particular immu- noglobulin See, e.g., Shapiro et al., Crit. Rev. Immunol. 22(3): 183-200 (2002); Marchalonis et al., Adv Exp Med Biol. 484:13-30 (2001 )).
- One of the advantages provided by various embodiments of the present invention stems from the finding that germline antibody genes are more likely than mature antibody genes are to conserve essential amino acid sequence structures characteristic of individuals in the species, hence less likely to be recognized as non-self when used in that species.
- key residues refers to certain residues within the variable region that have more impact on the binding specificity and/or affinity of an antibody, in particular a humanized antibody.
- a key residue includes, but is not limited to, one or more of the following: a residue that is adjacent to a CDR, a potential glycosylation site (which can be either N- or O- glycosylation site), a rare residue, a residue capable of interacting with the antigen, a residue capable of interacting with a CDR, a canonical residue, a contact residue between heavy chain variable region and light chain variable region, a residue within the Vernier zone, and a residue in the region that overlaps between the Chothia definition of a variable heavy chain CDR1 and the Kabat definition of the first heavy chain framework.
- the term “humanized antibody” specifically refers to an antibody or a variant, derivative, analog or fragment thereof which immunospecifically binds to an antigen of interest and which comprises a framework (FR) region having substantially the amino acid sequence of a human antibody and a complementary determining region (CDR) having substantially the amino acid sequence of a non-human antibody.
- FR framework
- CDR complementary determining region
- the term “substantially” in the context of a CDR refers to a CDR having an amino acid sequence at least 80%, preferably at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to the amino acid sequence of a non-human antibody CDR.
- a humanized antibody comprises substantially all of at least one, and typically two, variable domains (Fab, Fab', F(ab') 2, FabC, Fv) in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin (i.e., donor antibody) and all or substantially all of the framework regions are those of a human immunoglobulin consensus sequence.
- a humanized antibody also comprises at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin.
- a humanized antibody contains both the light chain as well as at least the variable domain of a heavy chain.
- the antibody also may include the CH1 , hinge, CH2, CH3, and CH4 regions of the heavy chain.
- a humanized antibody only contains a humanized light chain. In some embodiments, a humanized antibody only contains a humanized heavy chain. In specific embodiments, a humanized antibody only contains a humanized variable domain of a light chain and/or humanized heavy chain.
- the humanized antibody can be selected from any class of immunoglobulins, including IgM, IgG, IgD, IgA and IgE, and any subclass, including without limitation IgG 1 , lgG2, lgG3 and lgG4.
- the framework and CDR regions of a humanized antibody need not correspond precisely to the parental sequences, e.g., the donor antibody CDR or the consensus framework may be mutagenized by substitution, insertion and/or deletion of at least one amino acid residue so that the CDR or framework residue at that site does not correspond exactly to either the donor antibody or the consensus framework. In a preferred embodiment, such mutations, however, will not be extensive. Usually, at least 80%, preferably at least 85%, more preferably at least 90%, and most preferably at least 95% of the humanized antibody residues will correspond to those of the parental FR and CDR sequences.
- the term "consensus framework" refers to the framework region in the consensus immunoglobulin sequence.
- the term "consensus immunoglobulin sequence” refers to the sequence formed from the most frequently occurring amino acids (or nucleotides) in a family of related immunoglobulin sequences (see e.g., Winnaker, From Genes to Clones (Verlagsgesellschaft, Weinheim, Germany 1987). In a family of immunoglobulins, each position in the consensus sequence is occupied by the amino acid occurring most frequently at that position in the family. Where two amino acids occur equally frequently, either can be included in the consensus sequence.
- Vernier zone refers to a subset of framework residues that may adjust CDR structure and fine-tune the fit to antigen as described by Foote and Winter (1992, J. MoI. Biol. 224:487-499, which is incorporated herein by reference). Vernier zone residues form a layer underlying the CDRs and may impact on the structure of CDRs and the affinity of the antibody.
- epitope includes any polypeptide determinant capable of specific binding to an immunoglobulin.
- epitope determinants include chemically active surface groupings of molecules such as amino acids, sugar side chains, phosphoryl, or sulfonyl, and, in certain embodiments, may have specific three dimensional structural characteristics, and/or specific charge characteristics.
- An epitope is a region of an antigen that is bound by an antibody.
- an antibody is said to specifically bind an antigen when it preferentially recognizes its target antigen in a complex mixture of proteins and/or macromole- cules.
- polynucleotide as referred to herein means a polymeric form of two or more nucleotides, either ribonucleotides or deoxynucleotides or a modified form of either type of nucleotide.
- the term includes single and double stranded forms of DNA but preferably is double-stranded DNA.
- isolated polynucleotide shall mean a polynucleotide (e.g., of genomic, cDNA, or synthetic origin, or any combination thereof) that, by virtue of its origin , the "isolated polynucleotide” is not associated with all or a portion of a polynucleotide with which the "isolated polynucleotide” is found in nature; is operably linked to a polynucleotide that it is not linked to in nature; or does not occur in nature as part of a larger sequence.
- vector is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked.
- plasmid refers to a circular double stranded DNA into which additional DNA segments may be ligated.
- viral vector is another type of vector, wherein additional DNA segments may be ligated into the viral genome.
- Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors).
- vectors e.g., non-episomal mammalian vectors
- vectors can be integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome.
- certain vectors are capable of directing the expression of genes to which they are operatively linked.
- Such vectors are referred to herein as "recombinant expression vectors" (or simply, "expression vectors”).
- expression vectors of utility in recombinant DNA techniques are often in the form of plasmids.
- plasmid and vector may be used interchangeably as the plasmid is the most commonly used form of vector.
- the invention is intended to include such other forms of expression vectors, such as viral vectors (e.g., replication defective retroviruses, adenoviruses and adeno-associated viruses), which serve equivalent functions.
- operably linked refers to a juxtaposition wherein the components described are in a relationship permitting them to function in their intended manner.
- a control sequence "operably linked" to a coding sequence is connected in such a way that expression of the coding sequence is achieved under conditions compatible with the control sequences.
- "Operably linked” sequences include both expression control sequences that are contiguous with the gene of interest and expression control sequences that act in trans or at a distance to control the gene of interest.
- expression control sequence refers to polynucleotide sequences which are necessary to effect the expression and processing of coding sequences to which they are ligated.
- Expression control sequences include appropriate transcription initiation, termination, promoter and enhancer sequences; efficient RNA processing signals such as splicing and polyadenylation signals; sequences that stabilize cytoplasmic mRNA; sequences that enhance translation efficiency (i.e., Kozak consensus sequence); sequences that enhance protein stability; and when desired, sequences that enhance protein secretion.
- the nature of such control sequences differs depending upon the host organism; in prokaryotes, such control sequences generally include promoter, ribosomal binding site, and transcription termination sequence; in eukaryotes, generally, such control sequences include promoters and transcrip- tion termination sequence.
- control sequences is intended to include components whose presence is essential for expression and processing, and can also include additional components whose presence is advantageous, for example, leader sequences and fusion partner sequences.
- Transformation refers to any process by which exogenous DNA enters a host cell. Transformation may occur under natural or artificial conditions using various methods well known in the art. Transformation may rely on any known method for the insertion of foreign nucleic acid sequences into a prokaryotic or eukaryotic host cell. The method is selected based on the host cell being transformed and may include, but is not limited to, viral infection, electroporation, lipofection, and particle bombardment. Such "transformed” cells include stably transformed cells in which the inserted DNA is capable of replication either as an autonomously replicating plasmid or as part of the host chromosome. They also include cells which transiently express the inserted DNA or RNA for limited periods of time.
- host cell is intended to refer to a cell into which exogenous DNA has been introduced. It should be understood that such terms are intended to refer not only to the particular subject cell, but, also to the progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term "host cell” as used herein.
- host cells include prokaryotic and eukaryotic cells selected from any of the kingdoms of life. Preferred eukaryotic cells include protist, fungal, plant and animal cells.
- host cells include but are not limited to the prokaryotic cell line E.coli; mammalian cell lines CHO, HEK 293 and COS; the insect cell line Sf9; and the fungal cell Saccharomyces cerevisiae.
- Standard techniques may be used for recombinant DNA, oligonucleotide synthesis, and tissue culture and transformation (e.g., electroporation, lipofection).
- Enzymatic reactions and purification techniques may be performed according to manufacturer's specifications or as commonly accomplished in the art or as described herein.
- the foregoing techniques and procedures may be generally performed according to conventional methods well known in the art and as de- scribed in various general and more specific references that are cited and discussed throughout the present specification. See e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989)), which is incorporated herein by reference for any purpose.
- Transgenic organism refers to an organism having cells that contain a transgene, wherein the transgene introduced into the organism (or an ancestor of the organism) expresses a polypeptide not naturally expressed in the organism.
- a "transgene” is a DNA construct, which is stably and operably integrated into the genome of a cell from which a transgenic organism develops, directing the expression of an encoded gene product in one or more cell types or tissues of the transgenic organism.
- mice which had been made deficient in a particular endogenous protein via homologous recombination at the corresponding endogenous gene (i.e. knockout mice) have been shown to generate a humoral response to the protein with which they have been immunized and therefore to be able to be used for production of high-affinity monoclonal antibodies to the protein (see, for example, Roes, J. et al. (1995) J. Immunol. Methods 183:231-237; Lunn, MP. et al. (2000) J. Neurochem. 75:404-412).
- a multiplicity of nonhuman mammals are suitable hosts for antibody production in order to produce nonhuman antibodies of the invention. They include mice, rats, chickens, camelids, rabbits, sheep and goats (and knockout versions thereof), although preference is given to mice for the production of hybridomas. Furthermore, a nonhuman host animal expressing a human an- tibody repertoire may be used for producing essentially human antibodies to a human antigen with dual specificity. Nonhuman animals of this kind include transgenic animals (e.g. mice) bearing human immunoglobulin transgenes (chimeric hu-PBMC SCID mice) and human/mouse irradiation chimeras which are described in more detail below.
- transgenic animals e.g. mice
- human immunoglobulin transgenes chimeric hu-PBMC SCID mice
- human/mouse irradiation chimeras which are described in more detail below.
- the animal immunized with A ⁇ (20-42) globulomer or derivative thereof is a nonhuman mammal, preferably a mouse, which is transgenic due to human immunoglobulin genes so that said nonhuman mammal makes human antibodies upon antigenic stimulation.
- immunoglobulin transgenes for heavy and light chains with human germ line configuration are introduced into such animals which have been altered such that their endogenous heavy and light chain loci are inactive. If such animals are stimulated with antigen (e.g. with a human antigen), antibodies derived from the human immunoglobulin sequences (human antibodies) are produced. It is possible to make from the lymphocytes of such animals human monoclonal antibodies by means of standardized hybridoma technology.
- transgenic mice with human immunoglobulins and their use in the production of human antibodies, see, for example, US 5,939,598, WO 96/33735, WO 96/34096, WO 98/24893 and WO 99/53049 (Abgenix Inc.), and US 5,545,806, US 5,569,825, US 5,625,126, US 5,633,425, US 5,661 ,016, US 5,770,429, US 5,814,318, US 5,877,397 and WO 99/45962 (Genpharm Inc.); see also MacQuitty, J.J. and Kay, R.M. (1992) Science 257:1188; Taylor, L.D. et al. (1992) Nucleic Acids Res.
- the animal which is immunized with A ⁇ (20-42) globulomer or derivative thereof may be a mouse with severe combined immunodeficiency (SCID), which has been reconstituted with human peripheral mononuclear blood cells or lymphoid cells or precursors thereof.
- SCID severe combined immunodeficiency
- Such mice which are referred to as chimeric hu-PBMC SCID mice produce human immunoglobulin responses upon antigenic stimulation, as has been proved.
- the animal which is immunized with A ⁇ (20-42) globulomer or a derivative thereof is a mouse which has been treated with a lethal dose of total body irra- diation, then protected from radiation with bone marrow cells from mice with severe combined immunodeficiency (SCID) and subsequently transplanted with functional human lymphocytes.
- This type of chimera referred to as the Trimera system, is used in order to produce human monoclonal antibodies by immunizing said mice with the antigen of interest and then producing monoclonal antibodies by using standardized hybridoma technology.
- these mice and of their use for generating antibodies see, for example, Eren, R. et al.
- monoclonal antibodies may be produced by means of standardized techniques such as the hybridoma technique originally described by Kohler and Milstein (1975, Nature 256:495-497) (see also Brown et al. (1981 ) J. Immunol 127:539-46; Brown et al. (1980) J Biol Chem 255:4980-83; Yeh et al. (1976) PNAS 76:2927-31; and Yeh et al. (1982) Int. J. Cancer 29:269-75).
- the technology of producing monoclonal antibody hybhdomas is sufficiently known (see generally R. H.
- an immortalized cell line (typically a myeloma) is fused with lymphocytes (typically splenocytes or lymph node cells or peripheral blood lym- phocytes) of a mammal immunized with the A ⁇ globulomer of the invention or derivative thereof, and the culture supematants of the resulting hybridoma cells are screened in order to identify a hybridoma which produces a monoclonal antibody of the present invention.
- lymphocytes typically splenocytes or lymph node cells or peripheral blood lym- phocytes
- the immortalized cell line e.g. a myeloma cell line
- the immortalized cell line is derived from the same mammalian species as the lymphocytes.
- murine hybridomas may be established by fusing lymphocytes from a mouse immunized with an immunogenic preparation of the invention with an immortalized mouse cell line.
- Preferred immortalized cell lines are mouse myeloma cell lines which are sensitive to culture medium containing hypoxanthine, aminopterine and thymidine (HAT medium).
- myeloma cell lines may be used by default as fusion partner, for example the P3-NS1/1-Ag4-1 , P3- x63-Ag8.653 or Sp2/O-Ag14 myeloma lines.
- These myeloma cell lines are available from the American Type Culture Collection (ATCC), Rockville, MD.
- ATCC American Type Culture Collection
- HAT-sensitive mouse myeloma cells are fused to mouse splenocytes using polyethylene glycol (PEG).
- PEG polyethylene glycol
- Hybridoma cells resulting from the fusion are then selected using HAT medium, thereby killing unfused and unproductively fused myeloma cells (unfused splenocytes die after several days because they are not transformed).
- Hybridoma cells producing monoclonal antibodies of the invention are identified by screening the hybridoma culture supematants for such antibodies, for exam- pie by using a dot blot assay as described above and in example 8 in order to select those antibodies which have the binding affinities as defined above.
- the monoclonal antibodies 5F7, 10F11 , 7C6, 4B7, 6A2, 2F2, 4D10, 7E5, 10C1 , and 3B10 all have been generated using the above-described in-vivo approach and thereof are obtainable from a hybridoma as defined herein.
- said hybridoma can be used as a source of nucleic acid encoding light and/or heavy chains in order to recombinantly produce antibodies of the present invention, as is described below in further detail.
- anti- bodies of the invention may be identified and isolated by screening recombinant combinatorial immunoglobulin libraries with A ⁇ (20-42) globulomer or derivative thereof to thereby isolate immunoglobulin library members which have the required binding affinity.
- Kits for generating and screening display libraries are commercially available (e.g. the Pharmacia Recombinant Phage Antibody System, catalog No. 27-9400-01 ; and the Stratagene SurfZAP ® Phage Dis- play Kit, catalog No. 240612).
- the display library is an scFv library or an Fab library. The phage display technique for screening recombinant antibody libraries has been adequately described.
- Examples of methods and compounds which can be used particularly advantageously for generating and screening antibody display libraries can be found, for example, in McCafferty et al. WO 92/01047, US 5,969,108 and EP 589 877 (describes in par- ticular scFv display), Ladner et al. US 5,223,409, US 5,403,484, US 5,571 ,698, US 5,837,500 and EP 436 597 (describes pill fusion, for example); Dower et al. WO 91/17271 , US 5,427,908, US 5,580,717 and EP 527 839 (describes in particular Fab display); Winter et al.
- WO 97/29131 (describes production of a recombinant human antibody to a human antigen (human tumor necrosis factor alpha) and also in-vitro affinity maturation of the recombinant antibody) and Salfeld et al.
- U.S. Provisional Application No. 60/126,603 and the patent applications based hereupon (likewise describes production of recombinant human antibodies to human antigen (human interleukin-12), and also in-vitro affinity maturation of the recombinant antibody). Further descriptions of screenings of recombinant antibody libraries can be found in scientific publications such as Fuchs er a/. (1991 ) Bio/Technology 9: 1370-1372; Hay et al.
- recombinant antibody libraries may be expressed on the surface of yeast cells or of bacterial cells.
- WO 99/36569 describes methods of preparing and screening libraries expressed on the surface of yeast cells.
- WO 98/49286 describes in more detail methods of preparing and screening libraries expressed on the surface of bacterial cells.
- a selection process for enriching recombinant antibodies with the desired properties form an integral part of the process, which is generally referred to as "panning” and often takes the form of affinity chromatography over columns to whose matrix the target structure has been attached.
- Promising candidate molecules are then subjected to indi- vidual determination of their absolute and/or relative affinities, preferably by means of a standardized dot blot assay, as described above and in example 8.
- the DNA sequences encoding the light and heavy chains of said antibody are iso- lated by means of standardized molecular-biological techniques, for example by means of PCR amplification of DNA from the display package (e.g. the phage) which has been isolated during library screening.
- Nucleotide sequences of genes for light and heavy antibody chains, which may be used for preparing PCR primers, are known to the skilled worker. A multiplicity of such sequences are described, for example, in Kabat, E. A., er a/. (1991 ) Sequences of Pro- teins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242 and in the database of sequences of the human germ line VBASE.
- An antibody or antibody moiety of the invention may be produced by recombinantly expressing the genes for light and heavy immunoglobulin chains in a host cell.
- a host cell is transfected with one or more recombinant expression vectors carrying DNA fragments encoding the light and heavy immunoglobulin chains of said antibody, thereby expressing the light and heavy chains in the host cell and secreting them preferably into the medium in which said host cells are cultured.
- the antibodies can be isolated from this medium. Standardized recombinant DNA methods are used in order to obtain genes for heavy and light antibody chains, to insert said genes into recombinant expression vectors and to introduce said vectors into host cells.
- DNA fragments encoding VH and VL segments of the antibody of interest may be further manipulated using standardized recombinant DNA techniques, for example in order to convert the genes for variable regions to genes for full length antibody chains, to genes for Fab fragments or to an scFv gene.
- These manipulations comprise linking a VL- or VH-encoding DNA fragment operatively to another DNA fragment encoding another protein, for example a constant antibody region or a flexible linker.
- the term "operatively linked” is to be understood here as meaning that the two DNA fragments are linked in such a way that the amino acid sequences encoded by said two DNA fragments re- main in frame.
- the isolated DNA encoding the VH region may be converted to a gene for a full length heavy chain by operatively linking the VH-region encoding DNA with another DNA molecule encoding heavy chain constant regions (CH1 , CH2 and CH3).
- CH1 , CH2 and CH3 DNA molecule encoding heavy chain constant regions
- the sequences of human heavy chain constant region genes are well known (see, for example, Kabat, E. A., et al. (1991 ) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242), and DNA fragments spanning said regions may be obtained by means of standardized PCR amplification.
- the heavy chain constant region may be a constant region from IgGI , lgG2, lgG3, lgG4, IgM, IgA, IgE or IgD, with preference being given to a constant region from IgG, in particular IgGI or lgG4.
- the VH-encoding DNA may be operatively linked to another DNA molecule encoding merely the heavy chain constant region CH 1.
- the isolated DNA encoding the VL region may be converted to a gene for a full length light chain (and a gene for an Fab light chain) by operatively linking the VL-encoding DNA to another DNA molecule encoding the light chain constant region CL.
- the sequences of genes of the constant region of human light chain are well known (see Kabat, E.A., et al. (1991 ) Se- quences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242), and DNA fragments spanning said regions may be obtained by means of standardized PCR amplification.
- the light chain constant region may be a constant kappa or lambda region, a constant kappa region being preferred.
- the VH- and VL-encoding DNA fragments may be operatively linked to another fragment encoding a flexible linker, for example the amino acid se- quence (Gly 4 -Ser) 3 so that the VH and VL sequences are expressed as a continuous single- chain protein, with the VL and VH regions being linked to one another via said flexible linker (see Bird et al. (1988) Science 242:423-426; Huston et al. (1988) Proc. Natl. Acad. ScL USA 85:5879-5883; McCafferty et al., Nature (1990) 348:552-554).
- a flexible linker for example the amino acid se- quence (Gly 4 -Ser) 3 so that the VH and VL sequences are expressed as a continuous single- chain protein, with the VL and VH regions being linked to one another via said flexible linker
- Single domain VH and VL having the binding affinities as described above may be isolated from single domain libraries by the above-described methods.
- Two VH single-domain chains (with or without CH 1 ) or two VL chains or a pair of one VH chain and one VL chain with the desired binding affinity may be useful as described herein for the antibodies of the invention.
- the DNAs encoding partial or full length light and heavy chains may be inserted into expression vectors so as to operatively link the genes to appropriate transcriptional and translational control sequences.
- operatively linked is to be understood as meaning that an antibody gene is ligated in a vector in such a way that transcriptional and translational control sequences within the vector fulfill their intended function of regulating transcription and translation of said antibody gene.
- the expression vector and the expression control sequences are chosen so as to be compatible with the expression host cell used.
- the gene for the antibody light chain and the gene for the antibody heavy chain may be inserted into separate vectors or both genes are inserted into the same expression vector, this being the usual case.
- the antibody genes are inserted into the expression vector by means of standardized methods (for example by ligation of complementary restriction cleavage sites on the antibody gene fragment and the vector, or by ligation of blunt ends, if no restriction cleavage sites are present).
- the expression vector may already carry sequences for antibody constant regions prior to insertion of the sequences for the light and heavy chains.
- one approach is to convert the VH and VL sequences to full length antibody genes by inserting them into expression vectors already encoding the heavy and, respectively, light chain constant regions, thereby operatively linking the VH segment to the CH segment(s) within the vector and also operatively linking the VL segment to the CL segment within the vector.
- the recombinant expression vector may encode a signal peptide which facilitates secretion of the antibody chain from the host cell.
- the gene for said antibody chain may be cloned into the vector, thereby linking the signal peptide in frame to the N termi- nus of the gene for the antibody chain.
- the signal peptide may be an immunoglobulin signal peptide or a heterologous signal peptide (i.e. a signal peptide from a non-immunoglobulin protein).
- the expression vectors of the invention may have regulatory sequences controlling expression of the genes for the antibody chain in a host cell.
- regulatory sequence is intended to include promoters, enhancers and further expression control elements (e.g. polyadenylation signals) which control transcription or translation of the genes for the antibody chain. Regulatory sequences of this kind are described, for example, in Goeddel; Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, CA (1990). The skilled worker will appreciate that the expression vector design which includes selection of regulatory sequences may depend on factors such as the choice of the host cell to be transformed, the desired strength of expression of the protein, etc.
- Preferred regulatory sequences for expression in mammalian host cells include viral elements resulting in strong and constitutive protein expression in mammalian cells, such as promoters and/or enhancers derived from cytomegalovirus (CMV) (such as the CMV promoter/enhancer), simian virus 40 (SV40) (such as the SV40 promoter/enhancer), adenovirus (e.g. the adenovirus major late promoter (AdMLP)) and polyoma.
- CMV cytomegalovirus
- SV40 simian virus 40
- AdMLP adenovirus major late promoter
- the recombinant expression vectors of the invention may have additional sequences such as those which regulate replication of the vector in host cells (e.g. origins of replication) and selectable marker genes.
- the selectable marker genes facilitate the selection of host cells into which the vector has been introduced (see, for example, US patent Nos 4,399,216, 4,634,665 and 5,179,017, all to Axel et al.).
- Preferred selectable marker genes include the gene for dihydrofolate reductase (DHFR) (for use in dhfr host cells with methotrexate selection/amplification) and the neo gene (for G418 selection).
- DHFR dihydrofolate reductase
- neo gene for G418 selection.
- the expression vector(s) encoding said heavy and light chains is(are) transfected into a host cell by means of standardized techniques.
- the various forms of the term "transfection" are intended to comprise a multiplicity of techniques customarily used for introducing exogenous DNA into a prokaryotic or eukaryotic host cell, for example electroporation, calcium phosphate precipitation, DEAE-dextran transfection, and the like.
- the antibodies of the invention are expressed either in prokaryotic or eukaryotic host cells
- Prokaryotic expression of antibody genes has been reported as being ineffective for production of high yields of active antibody (Boss, M. A. and Wood, C. R. (1985) Immunology Today 6:12-13).
- Preferred mammalian host cells for expressing recombinant antibodies of the invention include CHO cells (including dhfr CHO cells described in Urlaub and Chasin, (1980) Proc. Natl. Acad. Sci. USA 77:4216-4220, which are used together with a DHFR-selectable marker, as described, for example, in R.J. Kaufman and P.A. Sharp (1982) MoI. Biol. 159:601-621 ), NSO myeloma cells, COS cells and SP2 cells.
- the antibodies When introducing recombinant expression vectors encoding the antibody genes into mammalian host cells, the antibodies are produced by cultur- ing the host cells until the antibody is expressed in said host cells or, preferably, the antibody is secreted into the culture medium in which the host cells grow. The antibodies may then be isolated from the culture medium by using standardized protein purification methods.
- host cells in order to produce moieties of intact antibodies, such as Fab fragments or scFv molecules. Variations of the above-described procedure are of course included in the invention. For example, it may be desirable to transfect a host cell with DNA encoding either the light chain or the heavy chain (but not both) of an antibody of the in- vention. If either light or heavy chains are present which are not required for binding of the antigen of interest, then the DNA encoding either such a light or such a heavy chain or both may be removed partially or completely by means of recombinant DNA technology. Molecules expressed by such truncated DNA molecules are likewise included in the antibodies of the invention.
- bifunctional antibodies in which a heavy chain and a light chain are an antibody of the invention and the other heavy chain and the other light chain have specificity for an antigen different from the antigen of interest, by crosslinking an antibody of the invention to a second antibody by means of standardized chemical methods.
- a recombinant expression vector encoding both the antibody heavy chain and the antibody light chain is introduced into dhfr CHO cells by means of calcium phosphate-mediated transfection.
- the genes for the heavy and light antibody chains are in each case operatively linked to regulatory CMV enhan- cer/AdMLP-promoter elements in order to effect strong transcription of said genes.
- the re- combinant expression vector also carries a DHFR gene which can be used for selecting dhfr CHO cells transfected with the vector by using methotrexate selection/amplification.
- the selected transformed host cells are cultured so that the heavy and light antibody chains are expressed, and intact antibody is isolated from the culture medium.
- Standardized molecular- biological techniques are used in order to prepare the recombinant expression vector, to trans- feet the host cells, to select the transformants, to culture said host cells, and to obtain the antibody from the culture medium.
- the invention relates to a method of synthesizing a recombinant antibody of the invention by culturing a host cell of the invention in a suitable culture medium until a recombinant antibody of the invention has been synthesized.
- the method may furthermore comprise isolating said recombinant antibody from said culture medium.
- any expression system in which a close physical linkage between a nucleic acid and the antibody encoded thereby is established and may be used to select a suitable nucleic acid sequence by virtue of the properties of the antibody it encodes may be employed.
- the recombinant antibody library is expressed in the form of RNA-protein fusions, as described in WO 98/31700 to Szostak and Roberts, and in Roberts, R.W. and Szostak, J.W. (1997) Proc. Natl. Acad. ScL USA 94:12297-12302.
- in-vitro translation of synthetic mRNAs carrying on their 3' end puromycin, a peptidyl acceptor antibiotic generates a covalent fusion of an mRNA and the peptide or protein encoded by it.
- a specific mRNA of a complex mixture of mRNAs e.g.
- a combinatorial library may be concentrated on the basis of the properties of the encoded peptide or protein (e.g. of the antibody or a moiety thereof), such as binding of said antibody or said moiety thereof to A ⁇ (20-42) globulomer or a derivative thereof.
- Nucleic acid sequences which encode antibodies or moieties thereof and which are obtained by screening of such libraries may be expressed by recombinant means in the above-described manner (e.g. in mammalian host cells) and may, in addition, be subjected to further affinity maturation by either screening in further rounds mRNA-peptide fusions, introducing mutations into the originally selected sequence ⁇ ), or using other methods of in-vitro affinity maturation of recombinant antibodies in the above-described manner.
- the antibodies of the invention may likewise be produced by using a combination of in-vivo and in-vitro approaches such as methods in which A ⁇ (20-42) globulomer or a derivative thereof is first allowed to act on an antibody repertoire in a host animal in vivo to stimulate production of A ⁇ (20-42) globulomer- or derivative-binding antibodies and then further antibody selection and/or antibody maturation (i.e. optimization) are accomplished with the aid of one or more in-vitro techniques.
- a combined method of this kind may comprise firstly immunizing a nonhuman animal (e.g.
- the first step of this combined procedure may be carried out in the manner described above in connection with the in-vivo approaches, while the second step of this procedure may be carried out in the manner described above in connection with the in-vitro ap- proaches.
- Preferred methods of hyperimmunizing nonhuman animals with subsequent in-vitro screening of phage display libraries prepared from said stimulated lymphocytes include those described by BioSite Inc., see, for example, WO 98/47343, WO 91/17271 , US 5,427,908 and US 5,580,717.
- a combined method comprises firstly immunizing a nonhu- man animal (e.g. a mouse, rat, rabbit, chicken, camelid, sheep, goat or a knockout and/or transgenic version thereof, or a chimeric mouse) with an A ⁇ (20-42) globulomer of the invention or derivative thereof to stimulate an antibody response to said A ⁇ (20-42) globulomer or derivative thereof and selecting the lymphocytes which produce the antibodies having the desired specificity by screening hybridomas (prepared, for example, from the immunized animals).
- a nonhu- man animal e.g. a mouse, rat, rabbit, chicken, camelid, sheep, goat or a knockout and/or transgenic version thereof, or a chimeric mouse
- the genes for the antibodies or single domain antibodies are isolated from the selected clones (by means of standardized cloning methods such as reverse transcriptase polymerase chain reaction) and subjected to in-vitro affinity maturation in order to improve thereby the binding properties of the selected antibody or the selected antibodies.
- the first step of this procedure may be conducted in the manner described above in connection with the in-vivo approaches, while the second step of this procedure may be conducted in the manner described above in connection with the in-vitro approaches, in particular by using methods of in-vitro affinity maturation, such as those described in WO 97/29131 and WO 00/56772.
- the recombinant antibodies are generated from individual isolated lymphocytes by using a procedure which is known to the skilled worker as selected lymphocyte antibody methods (SLAM) and which is described in US 5,627,052, WO 92/02551 and Babcock, J. S. et al. (1996) Proc. Natl. Acad. Sci. USA 93:7843-7848.
- SLAM selected lymphocyte antibody methods
- a non- human animal e.g.
- a mouse, rat, rabbit, chicken, camelid, sheep, goat, or a transgenic version thereof, or a chimeric mouse is firstly immunized in vivo with A ⁇ (20-42) globulomer or a derivative thereof to stimulate an immune response to said oligomer or derivative, and then individual cells secreting antibodies of interest are selected by using an antigen-specific haemo- lytic plaque assay.
- the globulomer or derivative thereof or structurally related molecules of interest may be coupled to sheep erythrocytes, using a linker such as biotin, thereby making it possible to identify individual cells secreting antibodies with suitable specificity by using the haemolytic plaque assay.
- variable regions of the light and heavy chains are obtained from the cells by reverse transcriptase PCR, and said variable regions may then be expressed in association with suitable immunoglobulin constant regions (e.g. human constant regions) in mammalian host cells such as COS or CHO cells.
- suitable immunoglobulin constant regions e.g. human constant regions
- the host cells transfected with the amplified immunoglobulin sequences derived from in vivo-selected lymphocytes may then be subjected to further in-vitro analysis and in-vitro selection by spreading out the transfected cells, for example, in order to isolate cells expressing antibodies with the binding affinity.
- the amplified immunoglobulin sequences may furthermore be manipulated in vitro.
- Antibodies having the required affinities defined herein can be selected by performing a dot blot essentially as described above. Briefly, the antigen is attached to a solid matrix, preferably dotted onto a nitrocellulose membrane, in serial dilutions. The immobilized antigen is then contacted with the antibody of interest followed by detection of the latter by means of an enzyme-conjugated secondary antibody and a colorimetric reaction; at defined antibody and antigen concentrations, the amount of antibody bound allows affinity determination.
- the relative affinity of two different antibodies to one target, or of one antibody to two different targets is here defined as the relation of the respective amounts of target-bound antibody observed with the two antibody-target combinations under otherwise identical dot blot conditions.
- Antibodies which bind to the same epitope as monoclonal antibody 5F7, 10F11 , 7C6, 4B7, 6A2, 2F2, 4D10, 7E5, 10C1 , or 3B10 can be obtained in a manner known per se.
- target structures are herein said to be "competing" for a particular antibody if at least one of these structures is capable of specifically reducing the measurable binding of another, preferably by offering an overlapping or identical epitope, more preferably an identical epitope.
- Competing target entities are useful for directly selecting antibodies by virtue of their relative affinity to such target structures.
- Relative affinities may thus be determined directly by using a competition assay in which distinguishable forms of the competing entities, e. g. differently labelled competing structures, are contacted with the antibody of interest, and the relative affinity of the antibody to each of these entities is deduced from the relative amounts of these entities which are bound by the antibody.
- Such competition may be used to directly enrich for antibodies possessing a desired relative affinity to the target entity, by attaching the entity towards which greater affinity is desired to a solid matrix support and adding a suitable amount, preferably a molar excess, of the competing entity towards which smaller affinity is desired to the medium.
- the antibodies display- ing the desired relative affinities will tend to bind to the matrix more strongly than others and may be obtained after washing out the less desirable forms, e. g. by washing out at low salt concentrations and then harvesting the bound antibody by reversibly detaching it from its target by using high salt concentrations.
- several rounds of enrichment may be performed.
- the genotype underlying an anti- body is physically linked to this antibody, e. g. in a pool of hybridomas or antigen-displaying phages or yeast cells, the corresponding phenotype may be rescued.
- a modified dot blot is used where the immobilized antigen competes with a solved entity for antibody binding, so that the relative affinity of the anti- body can be deduced from the percentage bound to the immobilized antigen.
- Antibody moieties such as Fab and F(ab') 2 fragments may be produced from whole antibodies by using conventional techniques such as digestion with papain or pepsin.
- antibodies, antibody moieties and immunoadhesion molecules may be obtained by using standardized recombinant DNA techniques.
- compositions comprising an antibody of the invention and, optionally, a pharmaceutically suitable carrier.
- Pharmaceutical compositions of the invention may furthermore contain at least one additional therapeutic agent, for example one or more additional therapeutic agents for the treatment of a disease for whose relief the antibodies of the invention are useful. If, for example, the antibody of the invention binds to a globulomer of the invention, the pharmaceutical composition may furthermore contain one or more additional therapeutic agents useful for the treatment of disorders in which the activity of said globulomer is important.
- Pharmaceutically suitable carriers include any solvents, dispersing media, coatings, antibacterial and antifungal agents, isotonic and absorption-delaying agents, and the like, as long as they are physiologically compatible.
- Pharmaceutically acceptable carriers include, for example, water, saline, phosphate-buffered saline, dextrose, glycerol, ethanol and the like, and combina- tions thereof.
- isotonic agents for example sugars, polyalcohols such as mannitol or sorbitol, or sodium chloride in addition.
- Pharmaceutically suitable carriers may furthermore contain relatively small amounts of auxiliary substances such as wetting agents or emulsifiers, preservatives or buffers, which increase the half life or efficacy of the antibodies.
- the pharmaceutical compositions may be suitable for parenteral administration, for example.
- the antibodies are prepared preferably as injectable solutions with an antibody content of 0.1 - 250 mg/ml.
- the injectable solutions may be prepared in liquid or lyophilized form, the dosage form being a flint glass or vial, an ampoule or a filled syringe.
- the buffer may contain L-histidine (1 - 50 mM, preferably 5 - 10 mM) and have a pH of 5.0 - 7.0, preferably of 6.0.
- Further suitable buffers include, without being limited thereto, sodium succinate, sodium citrate, sodium phosphate or potassium phosphate buffers.
- Sodium chloride may be used in order to adjust the tonicity of the solution to a concentration of 0 - 300 mM (preferably 150 mM for a liquid dosage form).
- Cryoprotectants for example sucrose (e.g. 0 - 10%, preferably 0.5 - 1.0%) may also be included for a lyophilized dosage form.
- Other suitable cryoprotectants are trehalose and lactose.
- Fillers for example mannitol (e.g. 1 - 10%, preferably 2 - 4%) may also be included for a lyophilized dosage form.
- Stabilizers for example L-methionine (e.g.
- 51 - 50 mM may be used both in liquid and lyophilized dosage forms.
- Further suitable fillers are glycine and arginine.
- Surfactants for example polysorbate 80 (e.g. 0 - 0.05%, preferably 0.005 - 0.01%), may also be used.
- Further surfactants are polysorbate 20 and BRIJ surfactants.
- compositions of the invention may have a multiplicity of forms. These include liquid, semi- solid and solid dosage forms, such as liquid solutions (e.g. injectable and infusible solutions), dispersions or suspensions, tablets, pills, powders, liposomes and suppositories.
- liquid solutions e.g. injectable and infusible solutions
- dispersions or suspensions tablets, pills, powders, liposomes and suppositories.
- the preferred form depends on the intended type of administration and on the therapeutic application. Typically, preference is given to compositions in the form of injectable or infusible solutions, for example compositions which are similar to other antibodies for passive immunization of hu- mans.
- the preferred route of administration is parenteral (e.g. intravenous, subcutaneous, intraperitoneal or intramuscular).
- the antibody is administered by intravenous infusion or injection.
- the antibody is administered by intramuscular or subcutaneous injection.
- compositions must typically be sterile and stable under preparation and storage conditions.
- the compositions may be formulated as solutions, microemulsions, dispersions, liposomes or other ordered structures suitable for high concentrations of active substance.
- Sterile injectable solutions may be prepared by introducing the active compound (i.e. the antibody) in the required amount into a suitable solvent, where appropriate with one or a combina- tion of the abovementioned ingredients, as required, and then sterile-filtering said solution.
- Dispersions are usually prepared by introducing the active compound into a sterile vehicle containing a basic dispersion medium and, where appropriate, other required ingredients.
- a sterile lyophilized powder for preparing sterile injectable solutions vacuum drying and spray drying are preferred methods of preparation, which produces a powder of the active ingredient and, where appropriate, of further desired ingredients from a previously sterile- filtered solution.
- the correct flowability of a solution may be maintained by using, for example, a coating such as lecithin, by maintaining, in the case of dispersions the required particle size or by using surfactants.
- a prolonged absorption of injectable compositions may be achieved by additionally introducing into the composition an agent which delays absorption, for example monostearate salts and gelatine.
- the antibodies of the invention may be administered by a multiplicity of methods known to the skilled worker, although the preferred type of administration for many therapeutic applications is subcutaneous injection, intravenous injection or infusion.
- the active compound may be prepared with a carrier which protects the compound against rapid release, such as, for example, a formulation with sustained or controlled release, which includes implants, transdermal plasters and microencapsulated release systems.
- a carrier which protects the compound against rapid release, such as, for example, a formulation with sustained or controlled release, which includes implants, transdermal plasters and microencapsulated release systems.
- Biologically degradable biocompatible polymers such as ethylene vinyl acetate, poly- anhydrides, polyglycolic acid, collagen, polyorthoesters and polylactic acid may be used.
- the methods of preparing such formulations are well known to the skilled worker; see, for example, Sustained and Controlled Release Drug Delivery Systems, J. R. Robinson, ed., Marcel Dekker, Inc., New York, 1978.
- an antibody of the invention may be administered orally, for example in an inert diluent or a metabolizable edible carrier.
- the antibody (and further ingredients, if desired) may also be enclosed in a hard or soft gelatine capsule, compressed to tablets or added directly to food.
- the antibodies may be mixed with excipients and used in the form of oral tablets, buccal tablets, capsules, elixirs, suspensions, syrups and the like. If it is intended to administer an antibody of the invention via a route other than the parenteral one, it may be necessary to choose a coating from a material which prevents its inactivation.
- the present invention also relates to a method of inhibiting the activity of globulomers of the invention in an individual which suffers from a disorder in which the amyloid ⁇ protein is involved and in which in particular the activity of said globulomers of the invention is important.
- Said method comprises the administration of at least one antibody of the invention to the individual with the aim of inhibiting the activity of the globulomer to which the antibody binds.
- Said individual is preferably a human being.
- An antibody of the invention may be administered for therapeutic purposes to a human individual.
- an antibody of the invention may be administered to a nonhuman mammal for veterinary purposes or within the framework of an animal model for a particular disorder. Such animal models may be useful for evaluating the therapeutic efficacy of antibodies of the invention (for example for testing dosages and time courses of administration).
- disorders in which the globulomers of the invention play a part include in particular disorders in whose development and/or progression a globulomer of the invention is involved. These are in particular those disorders in which globulomers of the invention are evidently or presumably responsible for the pathophysiology of said disorder or are a factor which contributes to the development and/or progression of said disorder. Accordingly, those disorders are included here in which inhibition of the activity of globulomers of the invention can relieve symptoms and/or progression of the disorder. Such disorders can be verified, for example, by an increased concentration of globulomers of the invention in a biological fluid of an individual suffering from a particular disorder (e.g. increased concentration in serum, plasma, CSF, urine, etc.).
- a particular disorder e.g. increased concentration in serum, plasma, CSF, urine, etc.
- disorders that can be treated or prevented include those associated with amyloidoses.
- amyloidoses here denotes a number of disorders characterized by abnormal folding, clumping, aggregation and/or accumulation of particular proteins (amyloids, fibrous proteins and their precursors) in various tissues of the body.
- ACA cerebral amyloid angiopathy
- compositions of the invention may include a "therapeutically effective amount” or a “prophylactically effective amount” of an antibody or antibody moiety of the inven- tion.
- a “therapeutically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result.
- a therapeutically effective amount of the antibody or antibody moiety may be determined by a person skilled in the art and may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the antibody or antibody moiety to elicit a desired response in the indi- vidual.
- a therapeutically effective amount is also one in which any toxic or detrimental effects of the antibody or antibody portion are outweighed by the therapeutically beneficial effects.
- prophylactically effective amount refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically, since a prophylactic dose is used in subjects prior to or at an earlier stage of disease, the prophylactically effective amount will be less than the therapeutically effective amount.
- the present invention includes a further method of preventing or treating Alzheimer's disease in a patient in need of such prevention or treatment.
- This method comprises the step of administering the vaccine noted above to the patient in an amount sufficient to effect the prevention or treatment.
- the present invention encompasses a method of identifying compounds suitable for active immunization of a patient predicted to develop an amyloidosis, e.g. Alzheimer's disease.
- This method comprises: 1 ) exposing one or more compounds of interest to one or more of the antibodies described above for a time and under conditions sufficient for the one or more compounds to bind to the antibody or antibodies; 2) identifying those compounds which bind to the antibody or antibodies, the identified compounds to be used in active immunization in a patient predicated to develop an amyloidosis, e.g. Alzheimer's disease.
- the antibodies of the inven- tion advantageously have detection threshold concentrations of less than 10 ng/ml of sample, preferably of less than 1 ng/ml of sample and particularly preferably of less than 100 pg/ml of sample, meaning that at least the concentration of globulomer per ml of sample, indicated in each case, advantageously also lower concentrations, can be detected by the antibodies of the invention.
- the detection is carried out immunologically. This may be carried out in principle by using any analytical or diagnostic assay method in which antibodies are used, including agglutination and precipitation techniques, immunoassays, immunohistochemical methods and immunoblot tech- niques, for example Western blotting or, preferably, dot blot methods. In vivo methods, for example imaging methods, are also included here.
- immunoassays are advantageous.
- Competitive immunoassays i.e. assays where antigen and labelled antigen (tracer) compete for antibody binding
- sandwich immunoas- says i.e. assays where binding of specific antibodies to the antigen is detected by a second, usually labelled antibody, are both suitable.
- assays may be either homogeneous, i.e. without separation into solid and liquid phases, or heterogeneous, i.e. bound labels are separated from unbound ones, for example via solid phase-bound antibodies.
- the various heterogeneous and homogeneous immunoas- say formats can be classified into particular classes, for example RIAs (radioimmunoassays), ELISA (enzyme-linked immunosorbent assay), FIA (fluorescence immunoassay), LIA (luminescence immunoassay), TRFIA (time-resolved FIA), IMAC (immunoactivation), EMIT (enzyme-multiplied immune test), TIA (turbidometric immunoassay), I-PCR (immuno-PCR).
- RIAs radioimmunoassays
- ELISA enzyme-linked immunosorbent assay
- FIA fluorescence immunoassay
- LIA luminescence immunoassay
- TRFIA time-resolved FIA
- IMAC immunoactivation
- EMIT enzyme-multiplied immune test
- TIA turbidometric immunoassay
- I-PCR immuno-PCR
- globulomer quantification of the invention preference is given to competitive immunoassays in which a defined amount of labelled globulomer derivative serving as tracer competes with the globulomer of the sample (containing an unknown amount of unlabelled globulomers) to be quantified for binding to the antibody used.
- the amount of antigen, i.e. the amount of globulomer, in the sample can be determined from the amount of the displaced tracer with the aid of a standard curve.
- enzymes have proved advantageous.
- Systems based on peroxidases in particular horseradish peroxidase, alkaline phosphatase and ⁇ -D- galactosidase, may be used, for example.
- Specific substrates whose conversion can be moni- tored photometrically, for example, are available for these enzymes.
- Suitable substrate systems are based on p-nitrophenyl phosphate (p-NPP), 5-bromo-4-chloro-3-indolyl phos- phate/nitroblue tetrazolium (BCIP/NPT), Fast-Red/naphthol-AS-TS phosphate for alkaline phosphatase; 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), o- phenylenediamine (OPT), 3,3',5,5'-tetramethylbenzidine (TMB), o-dianisidine, 5-aminosalicylic acid, 3-dimethylaminobenzoic acid (DMAB) and 3-methyl-2-benzothiazolinehydrazone (MBTH) for peroxidases; o-nitrophenyl- ⁇ -D-galactoside (o-NPG), p-nitrophenyl- ⁇ -D-galactoside and 4- methylumbellipheny
- the tracers used may be labelled globulomers. In this sense, a particular globulomer can be determined by labelling the globulomer to be determined and using it as tracer.
- the coupling of labels to globulomers for preparing tracers may be carried out in a manner known per se.
- the comments above on derivatization of globulomers of the invention are referred to by analogy.
- a number of labels appropriately modified for conjugation to proteins are available, for example biotin-, avidin-, extravidin- or streptavidin-conjugated enzymes, maleimide-activated enzymes and the like.
- These labels may be reacted directly with the oligomer or, if required, with the appropriately derivatized globulomer to give the tracer.
- a streptavidin-peroxidase conjugate is used, then this firstly requires biotinylation of the globulomer. This applies correspondingly to the reverse order. Suitable methods to this end are also known to the skilled worker.
- the antigen-antibody complex may be separated by binding it to the support, for example via an anti-idiotypical antibody coupled to said support, e.g. an antibody directed against rabbit IgG.
- an anti-idiotypical antibody coupled to said support, e.g. an antibody directed against rabbit IgG.
- Appropriate supports, in particular microtiter plates coated with appropriate antibodies, are known and partly commercially available.
- the present invention further relates to immunoassay sets having at least one antibody as described above and further components.
- Said sets are, usually in the form of a packaging unit, a combination of means for carrying out a globulomer determination of the invention.
- said means are preferably provided in an essentially ready-to-use form.
- An advantageous arrangement offers the immunoassay in the form of a kit.
- a kit usually comprises multiple containers for separate arrangement of components. All components may be provided in a ready-to-use dilution, as a concentrate for diluting or as a dry substance or lyophilisate for dissolving or suspending; individual or all components may be frozen or stored at room temperature until use.
- Sera are preferably shock-frozen, for example at -2O 0 C so that in these cases an immunoassay has to be kept preferably at temperatures below freezing prior to use.
- immunoassay Further components supplied with the immunoassay depend on the type of said immunoassay. Usually, standard protein, tracer which may or may not be required and control serum are supplied together with the antiserum. Furthermore, microtiter plates, preferably antibody-coated, buffers, for example for testing, for washing or for conversion of the substrate, and the enzyme substrate itself may also be included.
- the present invention also includes a method of diagnosing an amyloidosis, e.g. Alzheimer's disease, in a patient suspected of having this disease.
- This method comprises the steps of: 1 ) isolating a biological sample from the patient; 2) contacting the biological sample with at least one of the antibodies described above for a time and under conditions sufficient for formation of antigen/antibody complexes; and 3) detecting presence of the antigen/antibody complexes in said sample, presence of the complexes indicating a diagnosis of an amyloidosis, e.g. Alzheimer's disease, in the patient.
- the antigen may be, for example, an globulomer or a portion or fragment thereof which has the same functional properties as the full globulomer (e.g., binding activity).
- the present invention includes another method of diagnosing an amyloidosis, e.g. Alzheimer's disease in a patient suspected of having this disease.
- This method comprising the steps of: 1 ) isolating a biological sample from the patient; 2) contacting the biological sample with an antigen for a time and under conditions sufficient for the formation of antibody/antigen complexes; 3) adding a conjugate to the resulting antibody/antigen complexes for a time and under conditions sufficient to allow the conjugate to bind to the bound antibody, wherein the conjugate comprises one of the antibodies described above, attached to a signal generating compound capable of generating a detectable signal; and 4) detecting the presence of an antibody which may be present in the biological sample, by detecting a signal generated by the signal generating compound, the signal indicating a diagnosis of an amyloidosis, e.g. Alzheimer's disease in the patient.
- the antigen may be a globulomer or a portion or fragment thereof having the same functional properties as the full glob
- the present invention includes an additional method of diagnosing an amyloidosis, e.g. Alzheimer's disease in a patient suspected of having an amyloidosis, e.g. Alzheimer's disease.
- This method comprises the steps of: 1 ) isolating a biological sample from said patient; 2) contacting the biological sample with anti-antibody, wherein the anti-antibody is specific for one of the antibodies described above, for a time and under conditions sufficient to allow for formation of anti-antibody/antibody complexes, the complexes containing antibody present in the biological sample; 2) adding a conjugate to resulting anti-antibody/antibody complexes for a time and under conditions sufficient to allow the conjugate to bind to bound antibody, wherein the conjugate comprises an antigen, which binds to a signal generating compound capable of generat- ing a detectable signal; and 3) detecting a signal generated by the signal generating compound, the signal indicating a diagnosis of an amyloidosis, e.g. Alzheimer'
- the present invention includes a kit comprising: a) at least one of the antibodies described above and b) a conjugate comprising an antibody attached to a signal-generating compound, wherein the antibody of the conjugate is different from the isolated antibody.
- the present invention also encompasses a kit comprising: a) an anti-antibody to one of the antibodies described above and b) a conjugate comprising an antigen attached to a signal- generating compound.
- the antigen may be a globulomer or a fragment or portion thereof having the same functional characteristics as the globulomer (e.g., binding activity).
- an antibody of the present invention is coated on a solid phase (or is present in a liquid phase).
- the test or biological sample e.g., whole blood, cerebrospinal fluid, serum, etc.
- the test or biological sample e.g., whole blood, cerebrospinal fluid, serum, etc.
- antigen e.g., globulomer
- the direct method comprises simply detecting presence of the complex itself and thus presence of the antigens.
- a conjugate is added to the bound antigen.
- the conjugate comprises a second antibody, which binds to the bound antigen, attached to a signal- generating compound or label. Should the second antibody bind to the bound antigen, the signal-generating compound generates a measurable signal. Such signal then indicates presence of the antigen in the test sample.
- solid phases used in diagnostic immunoassays are porous and non-porous materials, latex particles, magnetic particles, microparticles (see U.S. Patent No. 5,705,330), beads, membranes, microtiter wells and plastic tubes.
- the choice of solid phase material and method of labeling the antigen or antibody present in the conjugate, if desired, are determined based upon desired assay format performance characteristics.
- the conjugate (or indicator reagent) will comprise an antibody (or perhaps anti-antibody, depending upon the assay), attached to a signal-generating compound or label.
- This signal-generating compound or label is itself detectable or may be reacted with one or more additional compounds to generate a detectable product.
- signal-generating compounds include chromogens, radioisotopes (e.g., 1251, 1311, 32P, 3H, 35S and 14C), chemiluminescent compounds (e.g., acridinium), particles (visible or fluorescent), nucleic ac- ids, complexing agents, or catalysts such as enzymes (e.g., alkaline phosphatase, acid phosphatase, horseradish peroxidase, beta-galactosidase and ribonuclease).
- chromo-, fluro-, or lumo-genic substrate results in generation of a detectable signal.
- detection systems such as time-resolved fluorescence, internal-reflection fluorescence, amplification (e.g., polymerase chain reaction) and Raman spectroscopy are also useful.
- the present invention also encompasses a method for detecting the presence of antibodies in a test sample.
- This method comprises the steps of: (a) contacting the test sample suspected of containing antibodies with anti-antibody specific for the antibodies in the patient sample under time and conditions sufficient to allow the formation of anti-antibody/antibody complexes, wherein the anti-antibody is an antibody of the present invention which binds to an antibody in the patient sample; (b) adding a conjugate to the resulting anti-antibody/antibody complexes, the conjugate comprising an antigen (which binds to the anti-antibody) attached to a signal generating compound capable of detecting a detectable signal; and (d) detecting the presence of the antibodies which may be present in the test sample by detecting the signal generated by the signal generating compound.
- a control or calibrator may be used which comprises antibody to the anti-antibody.
- kits are also included within the scope of the present invention. More specifically, the present invention includes kits for determining the presence of antigens (e.g., globulomers) in a patient suspected of having Alzheimer's disease or another condition characterized by cognitive impairment.
- a kit for determining the presence of antigens in a test sample comprises a) an antibody as defined herein or moiety thereof; and b) a conjugate comprising a second antibody (having specificity for the antigen) attached to a signal generating compound capable of generating a detectable signal.
- the kit may also contain a control or calibrator which comprises a reagent which binds to the antigen.
- the present invention also includes a kit for detecting antibodies in a test sample.
- the kit may comprise a) an anti-antibody specific (for example, one of the subject invention) for the anti- body of interest, and b) an antigen or portion thereof as defined above.
- a control or calibrator comprising a reagent which binds to the antigen may also be included.
- the kit may comprise a) an anti-antibody (such as the one of the present invention) specific for the antibody and b) a conjugate comprising an antigen (e.g., globulomer) attached to a signal generating compound capable of generating a detectable signal.
- the kit may also comprise a control of calibrator comprising a reagent which binds to the antigen.
- the kit may also comprise one container such as vial, bottles or strip, with each container with a pre-set solid phase, and other containers containing the respective conjugates.
- These kits may also contain vials or containers of other reagents needed for performing the assay, such as washing, processing and indicator reagents.
- the subject invention not only includes the full length antibodies described above but also moities or fragments thereof, for example, the Fab portion thereof. Additionally, the subject invention encompasses any antibody having the same properties of the present antibodies in terms of, for example, binding specificity, structure, etc.
- a response directed to vascular A ⁇ peptide in the form of CAA is to be avoided in order to eschew the undesirable side effect of microhaemorrhages (antibodies against the central portion of A ⁇ and which in addition do not bind to A ⁇ -peptides aggregated in the form of CAA induce fewer microhaemorrhages when compared to such against the N- terminus, see above).
- Antibodies which specifically react with A ⁇ oligomers will have higher bioavailability with regard to the pathophysiological ⁇ relevant A ⁇ species, as they will not be bound to, e.g., fibrillary A ⁇ and thus made unavailable for therapeutic effect.
- the hybridoma which produces monoclonal antibody 5F7 was deposited with the American Type Culture Collection, 10801 University Boulevard, Manassas, Virginia 20110 on December 01 , 2005 under the terms of the Budapest Treaty and received designa- tion PTA-7241. Further, the hybridoma which produces monoclonal antibody 10F11 was deposited with the American Type Culture Collection, 10801 University Boulevard, Manassas, Virginia 10801 on December 01 , 2005 under the terms of the Budapest Treaty and received designation PTA-7239.
- the hybridoma which produces monoclonal antibody 4B7 was deposited with the American Type Culture Collection, 10801 University Boulevard, Ma- nassas, Virginia 10801 on December 01 , 2005 under the terms of the Budapest Treaty and received designation PTA-7242, and the hybridoma which produces monoclonal antibody 7C6 was deposited with the American Type Culture Collection, 10801 University Boulevard, Manassas, Virginia 10801 on December 01 , 2005 under the terms of the Budapest Treaty and received designation PTA-7240.
- the hybridoma which produces monoclonal anti- body 6A2 was deposited with the American Type Culture Collection, 10801 University Boulevard, Manassas, Virginia 10801 on February 28, 2006 under the terms of the Budapest Treaty and received designation PTA-7409, and the hybridoma which produces monoclonal antibody 2F2 was deposited with the American Type Culture Collection, 10801 University Boulevard, Manassas, Virginia 10801 on February 28, 2006 under the terms of the Budapest Treaty and received designation PTA-7408.
- the hybridoma which produces monoclonal antibody 4D10 was deposited with the American Type Culture Collection, 10801 University Boulevard, Manassas, Virginia 10801 on February 28, 2006 under the terms of the Budapest Treaty and received designation PTA-7405.
- the hybridoma which produces monoclonal antibody 7E5 was deposited with the American Type Culture Collection, 10801 University Boulevard, Manassas, Virginia 10801 on August 16, 2006 under the terms of the Budapest Treaty and received des- ignation PTA-7809.
- the hybridoma which produces monoclonal antibody 10C1 was deposited with the American Type Culture Collection, 10801 University Boulevard, Manassas, Virginia 10801 on August 16, 2006 under the terms of the Budapest Treaty and received designation PTA-7810.
- the hybridoma which produces monoclonal antibody 3B10 was deposited with the American Type Culture Collection, 10801 University Boulevard, Manassas, Virginia 10801 on September 01 , 2006 under the terms of the Budapest Treaty and received designation PTA- 7851. All deposits have been made on behalf of Abbott Laboratories, 100 Abbott Park Road, Abbott Park, Illinois 60064 (US).
- Figure 1 shows size-exclusion chromatograms of A ⁇ (1-42) and A ⁇ (1-40).
- a ⁇ (1-42) monomer was dissolved in A) 0.1% NH 4 OH, B) 70% formic acid C) 0.1% NaOH and in D)
- a ⁇ (1-40) was dissolved in 0.1 % NaOH. Subsequently, the samples were further diluted 1 :10 in 2OmM NaH 2 PO 4 , 14OmM NaCI, pH 7.4 These samples were incubated for 5 min (left column) or 1 hour (right column) after dissolution at ambient temperature, then applied to the size exclusion column;
- Figure 2 A shows an SDS PAGE of standard proteins (molecular marker proteins, lane 1); A ⁇ (1-42) fibril preparation; control (lane 2); A ⁇ (1-42) fibril preparation + mAb
- Figure 3 is a bar diagram which shows the results of the object recognition test with
- APP/L transgenic mice after active immunization with A ⁇ (1-42) monomers in 0.1% NH 4 OH, A ⁇ (1-42) globulomers and A ⁇ (20-42) globulomers as compared to wild-type mice (positive control) and PBS-treated APP/L mice (negative control), where circles indicate significant differences to PBS-treated APP/L mice and asterisks indicate highly significant differences to chance level (50%) according to post-hoc t-test after P ⁇ 0.05 in ANOVA for differences among groups;
- Figure 4 shows dot blots of the reactivity of 100 pmol/ ⁇ l (row A); 10 pmol/ ⁇ l (row B); 1 pmol/ ⁇ l (row C); 0.1 pmol/ ⁇ l (row D) and 0.01 pmol/ ⁇ l (row E) of A ⁇ (1-42) globulomer (column 1); of HFIP pretreated A ⁇ (1-42) monomer in Pluronic F68 (column 2); of A ⁇ (20-42) globulomer (column 3); of A ⁇ (12-42) globulomer (column 4); of HFIP pretreated A ⁇ (1-40) monomer in DMSO (column 5); of A ⁇ (1- 42) monomer, NH 4 OH (column 6); of an A ⁇ (1-42) fibril preparation (column 7); and of sAPP ⁇ from Sigma (column 8) with various antisera obtained after an active immunization of APP/L Tg mice with A ⁇ (20-4
- Figure 5 is a bar diagram which shows the concentrations of soluble and insoluble A ⁇ (1- 42) and A ⁇ (1-40) peptide in brain extracts of actively immunized APP/PS1 Tg- mice with either A ⁇ (1 -42) monomer (0.1 % NH 4 OH), A ⁇ (1 -42) globulomer, A ⁇ (20-42) globulomer or vehicle as control;
- Figure 6 is a bar diagram which shows the results of the object recognition test with
- Figure 7 A shows a dot blot analysis of the specificity of different anti-A ⁇ antibodies
- the monoclonal antibodies tested here were obtained by active immunization of mice with A ⁇ (20-42) globulomer followed by selection of the fused hybridoma cells (except for the commercial available 6E10, Signet No 9320). The individual A ⁇ forms were applied in serial dilutions and incubated with the respective monoclonal antibodies for immune reaction:
- a ⁇ (1-42) monomer 0.1 % NH 4 OH 2.
- a ⁇ (1-40) monomer 0.1% NH 4 OH
- FIG. 9 Anti-A ⁇ -antibody titer and dot-blot selectivity profile in plasma of TG2576 mice approximately one year after active immunization. Plasma samples of Tg2576- mice approximately one year after the last immunization with A) A ⁇ (20-42) globulomer, B) A ⁇ (12-42) globulomer, C) A ⁇ (1-42) monomer and D) vehicle, were assessed for anti-A ⁇ antibodies produced and still present by dot-blot.
- Figure 10 shows a table summarizing the levels of A ⁇ (20-42) globulomer in brain tissue of human beings having Alzheimer's disease and a non demented control;
- Figure 11 illustrates nucleotide and amino acid sequences of the variable heavy and light chains of monoclonal antibodies (mAbs) as follows (complementarity determining regions (CDRs) are underlined in each amino acid sequence):
- the A ⁇ (1-42) synthetic peptide (H-1368, Bachem, Bubendorf, Switzerland) was suspended in 100% 1 ,1 ,1 ,3,3,3-hexafluoro-2-propanol (HFIP) at 6 mg/mL and incubated for complete solubilization under shaking at 37 0 C for 1.5 h.
- the HFIP acts as a hydrogen-bond breaker and is used to eliminate pre-existing structural inhomogeneities in the A ⁇ peptide.
- HFIP was removed by evaporation in a Speed Vac and A ⁇ (1-42) resuspended at a concentration of 5 m M in di- methylsulfoxide and sonicated for 20 s.
- the HFIP-pre-treated A ⁇ (1-42) was diluted in phosphate-buffered saline (PBS) (20 mM NaH 2 PO 4 , 140 mM NaCI, pH 7.4) to 400 ⁇ M and 1/10 volume 2% sodium dodecyl sulfate (SDS) (in H 2 O) added (final concentration of 0.2% SDS).
- PBS phosphate-buffered saline
- SDS sodium dodecyl sulfate
- the 38/48-kDa A ⁇ (1-42) globulomer was generated by a further dilution with three volumes of H 2 O and incubation for 18 h at 37 0 C. After centrifugation at 3000 g for 20 min the sample was concentrated by ultrafiltration (30-kDa cut-off), dialysed against 5 mM NaH 2 PO 4 , 35mM NaCI, pH 7.4, centrifuged at 10000 g for 10 min and the supernatant comprsing the 38/48-kDa A ⁇ (1-42) globulomer withdrawn.
- the 38/48-kDa A ⁇ (1-42) globulomer could also be precipitated by a ninefold excess (v/v) of ice- cold methanol/acetic acid solution (33% methanol, 4% acetic acid) for 1 h at 4 ° C.
- the 38/48- kDa A ⁇ (1-42) globulomer is then pelleted (10 min at 16200 g), resuspended in 5 mM NaH 2 PO 4 , 35 mM NaCI, pH 7.4, and the pH adjusted to 7.4.
- the A ⁇ (1-42) synthetic peptide (H-1368, Bachem, Bubendorf, Switzerland) was suspended in 100% 1 ,1 ,1 ,3,3,3-hexafluoro-2-propanol (HFIP) at 6 mg/mL and incubated for complete solubi- lization under shaking at 37 0 C for 1.5 h.
- the HFIP acts as a hydrogen-bond breaker and was used to eliminate pre-existing structural inhomogeneities in the A ⁇ peptide.
- HFIP was removed by evaporation in a SpeedVac and A ⁇ (1-42) resuspended at a concentration of 5 mM in di- methylsulfoxide and sonicated for 20 s.
- the HFIP-pre-treated A ⁇ (1-42) was diluted in phosphate-buffered saline (PBS) (20 mM NaH 2 PO 4 , 140 mM NaCI, pH 7.4) to 400 ⁇ M and 1/10 volume 2% sodium dodecyl sulfate (SDS) (in H 2 O) added (final concentration of 0.2% SDS).
- PBS phosphate-buffered saline
- SDS sodium dodecyl sulfate
- the 38/48-kDa A ⁇ (1-42) globulomer was generated by a further dilution with three volumes of H 2 O and incubation for 18 h at 37 0 C.
- Cross-linking of the 38/48-kDa A ⁇ (1-42) globulomer was now performed by incubation with 1 mM glutaraldehyde for 2 h at 21 0 C room temperature (RT) followed by ethanolamine (5 mM) treatment for 30 min at RT.
- the concentrate was ad- mixed with 9 ml of buffer (50 mM MES/NaOH, 0.02 % SDS, pH 7.4) and again concentrated to 1 ml.
- the concentrate was dialyzed at 6°C against 1 I of buffer (5 mM sodium phosphate, 35 mM NaCI) in a dialysis tube for 16 h.
- the dialysate was adjusted to an SDS content of 0.1% with a 2% strength SDS solution in water.
- the sample was centrifuged at 10000 g for 10 min and the A ⁇ (20-42) globulomer supernatant was withdrawn.
- Example 2 Size-exclusion chromatography of different A ⁇ (1-42) monomer and A ⁇ (1-40) monomer preparations
- a ⁇ (1-42), 0.1% NH 4 OH: 1 mg of A ⁇ (1-42) (Bachem, catalogue no. H-1368) were dissolved in 500 ⁇ l of 0.1% NH 4 OH in H 2 O and agitated for 1 min at ambient temperature. The sample was centrifuged for 5 min at 10'0OO g. The supernatant was collected. A ⁇ (1-42) concentration in the supernatant was determined according to Bradford's method (BIO-RAD).
- a ⁇ (1-42), 70% HCOOH 1 mg of A ⁇ (1-42) were dissolved in 50 ⁇ l 70% HCOOH in H 2 O and agitated 1 min at ambient temperature. The sample was centrifuged for 5 min at 10'0OO g. The supernatant was collected. A ⁇ (1-42) concentration in the supernatant is determined according to Bradford's method (BIO-RAD).
- a ⁇ (1-42) (Bachem, catalogue no. H-1368) were dissolved in 500 ⁇ l of 0.1% NaOH in H 2 O and agitated 1 min at ambient temperature. The sample was centrifuged for 5 min at 10'0OO g. The supernatant was collected. A ⁇ (1-42) concentration in the supernatant is determined according to Bradford's method (BIO-RAD).
- a ⁇ ( 1-40) (Bachem, catalogue no. H-1 194) were dissolved in 500 ⁇ l of 0.1 % NaOH in H 2 O and agitated 1 min at ambient temperature. The sample is was centrifuged for 5 min at 10'0OO g. The supernatant was collected. A ⁇ (1-42) concentration in the supernatant was determined according to Bradford's method (BIO-RAD).
- sample 20 ⁇ l of A ⁇ (1-40) in 0.1 % NaOH were diluted to a concentration of 0.2 mg/ml A ⁇ (1-40) with 20 mM NaH 2 PO 4 , 140 mM NaCI, pH 7.4. The sample was incubated for 5 min at ambient temperature. Then 100 ⁇ l were analyzed by size exclusion chromatography.
- SEC size exclusion chromatography
- the A ⁇ peptide supplier states in their technical information that the A ⁇ (1-42) should be solubilized in 0.1 % NH 4 OH.
- Monomeric A ⁇ (1-42) runs at a major peak with 11 kD and a shoulder at 6kD.
- a ⁇ (1-42) The best initial solvent for A ⁇ (1-42) to prevent aggregation is 0.1% NaOH which even after 1 h incubation of solubilization and further dilution shows only a minor fraction of aggregated A ⁇ (1-42) with the majority of A ⁇ (1-42) being still monomeric.
- a ⁇ (1-40) solubilized initially in 0.1% NaOH shows no signs at all of aggregation even after 1 h at RT incubation.
- Example 3 Semi-quantitative analysis visualized by SDS-PAGE of the discrimination of A ⁇ (20- 42) globulomer selective antibodies for A ⁇ (1-42) fibrils.
- a ⁇ (1-42) (Bachem, Cat. no.: H-1368) were dissolved in 500 ⁇ l 0.1% NH 4 OH in H 2 O and agitated for 1 min at ambient temperature. The sample was centrifuged for 5 min at 10'0OO g. The supernatant was collected. A ⁇ (1-42) concentration in the supernatant was determined according to Bradford's method (BIO-RAD).
- a ⁇ (1-42) fibril preparation 40 ⁇ l of A ⁇ (1-42) fibril preparation were diluted with 160 ⁇ l of 20 mM NaH 2 PO 4 , 140 mM NaCI, 0.05% Tween 20, pH 7.4 and agitated 5 min at ambient temperature, then the sample was centrifuged for 10 min at 10'0OO g. The supernatant was discarded, and the residue was resuspended in 95 ⁇ l of 20 mM NaH 2 PO 4 , 140 mM NaCI, 0.05% Tween 20, pH 7.4. Resuspen- sion was prompted by vigorous agitation ("vortexing").
- the samples were incubated at 37 0 C for 20 hours, then centrifuged for 10 min at 10'0OO g.
- the supernatants were collected and mixed with 20 ⁇ l of SDS-PAGE sample buffer.
- the residues were mixed with 50 ⁇ l of 20 mM NaH 2 PO 4 , 140 mM NaCI, 0.025% Tween 20, pH 7.4 and resuspended by "vortexing", then the samples were centrifuged for 10 min at 10'0OO g.
- the supernatants were discarded, and the residues were mixed with 20 ⁇ l 20 mM NaH 2 PO 4 , 140 mM NaCI, 0.025% Tween 20, pH 7.4, then with 20 ⁇ l of SDS-PAGE sample buffer.
- the samples were applied to a 4 - 20% Tris/glycine gel for electrophoresis.
- Tris/Glycine Gel (Invitrogen, Cat. no.: EC6025BOX)
- Electrophoresis buffer 7.5 g Tris
- the gel is run at a constant current of 20 mA.
- a ⁇ (1 -42) fibrils Semiquantitative analysis of different anti-A ⁇ antibodies and their discrimination of A ⁇ (1 -42) fibrils. Positions of antibodies, A ⁇ (1-42) fibrils and A ⁇ (1-42) monomers are marked at the edge of the gel. Due to their size, A ⁇ (1-42) fibrils cannot enter the SDS-PAGE gel and can be seen in the gel slot.
- a ⁇ (1-42) fibril preparation + mAb 6E10; 2Oh 37°C; supernatant 6.
- the relative binding to fibril type A ⁇ was evaluated from SDS-PAGE analysis by measuring the Optical Density (OD) values from the Heavy Chain of the antibodies in the fibril bound (pellet- fraction) and the supernatant fractions after centrifugation.
- OD Optical Density
- Antibodies that have bound to the A ⁇ fibrils should be co-pelleted with the A ⁇ -fibrils and therefore are found in the pellet fraction whereas non-A ⁇ -fibril bound (free) antibodies are found in the supernatant.
- the percentage of antibody bound to A ⁇ -fibrils was calculated according to the following formula:
- Percent antibody bound to A ⁇ -fibrils ODfjb ⁇ i fraction ⁇ 100%/(ODf ⁇ b ⁇
- a ⁇ fibrils are a major component of the total A ⁇ peptide pool.
- a ⁇ -antibodies By attacking these fibrils by anti A ⁇ -antibodies the risk of negative side effects is elevated due to a liberation of high amounts of A ⁇ which subsequently may increase the risk of micro- haemorrhages.
- An increased risk for microhaemorrhages was observed in an active immunization approach with fibrillar aggregates of the A ⁇ peptide (Bennett and Holtzman, 2005, Neurol- ogy, 64, 10-12; Orgogozo J, Neurology, 2003, 61 ,46-54; Schenk et al., 2004, Curr Opin Immunol, 16, 599-606).
- the A ⁇ (20-42) globulomer selective antibody 5F7 (which actually has the lowest selectivity for A ⁇ (20-42) globulomers over other A ⁇ -forms) does not bind to A ⁇ (1-42) fibrils in an co-pelleting experiment. This is shown by the fact that the 5F7 antibody after an incubation with A ⁇ (1-42) fibrils remains after a pelleting step in the supernatant and is not co-pelleted because of being bound to the A ⁇ (1-42) fibrils.
- Example 4 Analysis of cognitive performance in mice by means of an object recognition test after active immunization with A ⁇ (1-42) monomer (0.1% NH 4 OH), A ⁇ (1-42) globulomer or A ⁇ (20-42) globulomer in comparison to wild type.
- mice overexpressing human APP with a point mutation were used.
- the point mutation refers to amino acid 717 (substitution of isoleucine for valine) and has been found in a London family where it leads to onset of AD before the beginning of the sixth decade of life (Mullan et al., Nature Genetics 2 (1992) 340-342).
- the transgenic mice, herein referred to as APP/L were created by and first described in Leuven (Moechars et al., J. Biol. Chem. 274 (1999) 6483-6492). Female APP/L mice were subjected to active immunization at 6 weeks of age.
- mice received either 100 ⁇ g of A ⁇ (1-42) monomer (0.1 % NH 4 OH), A ⁇ (1-42) globulomer or A ⁇ (20-42) globulomer in phosphate-buffered saline (PBS) mixed with an equal amount of complete Freund's adjuvant intraperitoneally, followed by booster injections with the same amount of antigene in incomplete Freund's adjuvant every third week for three months.
- PBS phosphate-buffered saline
- the ..recognition index expresses this relation (time for new object/total time).
- a mouse which does not remember the known object will consider it as equally interesting as the new object and spend an equal amount of time on exploring it, in other words, will show a recognition index of 50%.
- a mouse which remembers the known object will consider it as not interesting and therefore show a significantly higher recognition index.
- APP/L mice are known to be cognitively deficient at 4.5 months of age and exhibit a recognition index in the dimension of the random level, i.e. 50%.
- mice Object recognition test in mice.
- the test reports recognition of a known object in comparison to an unknown one, measured in terms of explorative behaviour during a 10 minute test phase.
- the recognition index is defined as the percentage of time which the mouse spends on explor- ing the unknown object relative to the time spent on exploring both objects.
- the known object was explored by the mouse during a 10 minute acquisition phase three hours before the test phase.
- the other four groups of APP transgenic mice were subjected to active immunisation three months before.
- the immunogens used were A ⁇ (1-42) monomer, A ⁇ (1-42) globulomer and A ⁇ (20-42) globulomer.
- APP/L mice are known to show, in contrast to non-transgenic mice, a cognitive deficiency at 4.5 months of age, scoring results close to random level (i.e. 50% recognition index).
- mice showed random behaviour, in contrast to non-transgenic mice (wild type). Immunization with native A ⁇ (1-42) globulomer as well as with A ⁇ (20-42) globulomer resulted in significantly improved object recognition in APP/L mice.
- Example 5 Dot-Blot analysis of the antibody profile for different A ⁇ -forms after an active immunization of APP/L Tg mice with A ⁇ (20-42) globulomer.
- mice were assessed for anti-A ⁇ antibodies.
- dilution series of the individual A ⁇ (1-42) forms ranging from 100 pmol/ ⁇ l to 0.01 pmol/ ⁇ l in PBS supplemented with 0.2 mg/ml BSA were made. 1 ⁇ l of each sample was blotted onto a nitrocellulose membrane. For detection the corresponding mouse plasma samples were used (diluted 1 :400). lmmunostaining was done using alkaline phosphatase conjugated anti-mouse-lgG and the staining reagent NBT/BCIP.
- a ⁇ (20-42) globulomer The preparation of the A ⁇ (20-42) globulomer is described in example 1c.
- a ⁇ (1-42) (Bachem Inc. Catalog Nr.: H-1368) were dissolved in 500 ⁇ l aqueous 0.1% NH 4 OH (Eppendorff tube) and the sample was stirred for 1 min at room temperature. 100 ⁇ l of this freshly prepared A ⁇ (1-42) solution were neutralized with 300 ⁇ l 20 mM NaH 2 PO 4 ; 140 mM NaCI, pH 7.4. The pH was adjusted to pH 7.4 with 1% HCI. The sample was incubated for 24 h at 37°C and centrifuged (10 min at 1000Og). The supernatant was discarded and the fibril pellet resuspended with 400 ⁇ l of 20 mM NaH 2 PO 4 ; 140 mM NaCI, pH 7.4 by vortexing for 1 min.
- BSA Bovine Serum Albumin
- Antibody solution I Mouse plasma samples from an active immunization study with A ⁇ (20- 42) globulomer (1 :400 diluted in 20 ml 1 % low fat milk in TBS)
- Antibody solution II 1 :5000 dilution
- the dot blot was incubated with 30 ml 5 % low fat milk in TBS for 1.5 h at RT.
- the washing buffer was discarded and the dot blot was incubated with antibody solu- tion I overnight at RT.
- the antibody solution I was discarded and the dot blot was incubated under shaking with 20 ml TTBS for 10 min at RT.
- the washing solution was discarded and the dot blot was incubated under shaking with 20 ml TTBS for 10 min at RT.
- the washing solution was discarded and the dot blot was incubated under shaking with 20 ml TBS for 10 min at RT.
- Antibody solution II The washing buffer was discarded and the dot blot was incubated with antibody solution Il for 1 h at RT
- Plasma samples from APP/L Tg mice after being actively immunized with A ⁇ (20-42) globulomer exhibit an antibody profile (predominant recognition of A ⁇ (20-42) globulomer and A ⁇ (12-42) globulomer) which resembles that of the A ⁇ (20-42) globulomer mAbs claimed herein.
- Example 6 Concentration of soluble and insoluble A ⁇ (1-42) and A ⁇ (1-40) peptide in brain extracts of actively immunized APP/PS1 Tg mice with either A ⁇ (1-42) monomer, A ⁇ (1-42) globulomer, A ⁇ (20-42) globulomer or vehicle as control.
- 40 female mice of a double transgenic mouse model of Alzheimer's Disease (APP/PS1 Tg mice) in FVBxC57BI/6J background with an age of 4 months were used for this study.
- the APP/PS1 Tg mice contain the 695 amino acid form of human APP with the V717I mutation (position refering to the longest APP-isoform) and in addition the human Presenilin 1 gene with the A264E mutation.
- mice were generated and characterized in one of the founding laboratories of reMYND, the Experimental Genetics Group, Campus Gasthuisberg, Catholic University Leuven, by Prof. Fred Van Leuven et al.
- mice were genotyped by polymerase chain reaction (PCR) at the age of 3 weeks and received a unique identity number, once the PCR results were known.
- PCR polymerase chain reaction
- mice had free access to pre-filtered and sterile water (UV-lamp) and standard mouse chow.
- the food was stored under dry and cool conditions in a well-ventilated storage room. The amount of water and food was checked daily, supplied when necessary and by default refreshed twice a week.
- mice were housed under a reversed day-night rhythm: 14 hours light/10 hours darkness starting at 7 p.m., in standard metal cages type RVS T2 (area of 540 cm 2 ). The cages are equipped with solid floors and layer of bedding litter. The number of mice per cage was limited in accordance with legislation on animal welfare. Five days before the onset of the behaviour test, mice were re-caged in macrolon Type 2 cages and transported to the laboratory in order to adapt to the laboratory environment in preparation to the behaviour test.
- mice received either 100 ⁇ g of A ⁇ (1-42) monomer (0.1% NH 4 OH), A ⁇ (1-42) globulomer or A ⁇ (20-42) globulomer in phosphate-buffered saline (PBS) mixed with an equal amount of complete Freund's adjuvant intraperitoneally, followed by booster injections with the same amount of antigene in incomplete Freund's adjuvant every third week for four months.
- PBS phosphate-buffered saline
- the A ⁇ (1-40) and A ⁇ (1-42) in the soluble fraction of 1 hemisphere was determined by ELISA.
- mice were anaesthetized with a 2:1 :1 mixture of Ketalar (ketamin), Rompun (xylazin 2%) and atropin and flushed trans-cardially with physiological serum at 4 0 C. This was performed to remove blood from the brain vessels, a procedure which has no influence on organ integrity. Cerebrospinal fluid (CSF) was collected via an incision in the neck muscles, between the skull and the first cervical vertebrae. A puncture into the cistema magna was given with a 26 gauge needle and 10-20 ⁇ l of CSF was collected with a fine glass pipette.
- CSF Cerebrospinal fluid
- Blood was collected via a heart puncture and a 1 ml syringe into heparin-coated Eppendorf tubes. The blood was centrifuged at 14.000 rpm at 4 0 C for 5 minutes. The serum was stored at -70 0 C.
- mice were flushed transcardially with physiological serum at 4 0 C.
- the brain was removed from the cranium and hindbrain and forebrain were separated by a cut in the coronal/frontal plane.
- the cerebellum was discharged.
- the forebrain was divided evenly into left and right hemisphere by using a midline sagittal cut.
- Brains were homogenized using a Potter, a glass tube (detergent free, 2 cm 3 ) and a mechanical homogenizer (650 rpm). A volume of 6,5 x ⁇ A brain weight of freshly prepared 20 mM Tris/HCI buffer (pH 8.5) with Proteinase Inhibitors (1 tablet per 50 ml Tris/HCI buffer, CompleteTM, Roche, Mannheim, Germany) was used as homogenization buffer.
- the supernatant (soluble fraction containing secreted APP and amyloid peptides) was separated from the pellet (membrane fraction containing membrane-bound APP-fragments and plaque-associated amyloid peptides in case of aged mice). The supernatant was divided over two tubes of which one was stored at -2O 0 C as back-up and the other was processed further for column chromatography to concentrate the amyloid peptides.
- Brain weights, volume Tris/HCI buffer used, centrifugation sessions (marked by colour) and volume soluble fraction used for column chromatography are given exemplary in the following table.
- the pellets were further fractionated into different membrane fractions: membrane fraction A (MFA), membrane fraction B (MFB) containing full length APP and membrane fraction C (MFC) containing plaque associated amyloid. Therefore the pellets were dissolved in TBS buffer with proteinase inhibitors (1 tablet per 50 ml TBS buffer, CompleteTM, Roche, Mannheim, Germany) and the MFA was divided over two tubes of which one was stored at -20°C as backup.
- MFA membrane fraction A
- MFB membrane fraction B
- MFC membrane fraction C
- Brain weights, 60% of the brain weight, the volumes TBS + Pl + NP40 + Triton X-100 buffer used and the centrifugation sessions (marked by colour) are given exemplary in the following table.
- ELISA Enzyme- Linked-lmmunosorbent-Assay
- Samples, standards and blanks (50 ⁇ l) were added to the anti-A ⁇ -coated polystyrol plate (capture antibody selectively recognizes the C-terminal end of the antigen) in addition with a selec- tive anti-A ⁇ -antibody conjugate (biotinylated detection antibody) and incubated overnight at 4°C in order to allow formation of the antibody-Amyloid-antibody-complex.
- a Streptavidine-Peroxidase-Conjugate was added, followed 30 minutes later by the addition of a TMB/peroxide mixture, resulting in the conversion of the substrate into a coloured product.
- the MFC samples were further processed and dissolved in 8M Guanidine in 80 mM Tris/HCI. Subsequently samples were incubated for 3 hours in a ther- momixer at 25 0 C and pipetted up and down with a 100 ⁇ l pipette every hour to dissolve the MFC pellet into the guanidine buffer. Finally samples were centrifuged for only 1 minute at 4000 rpm to remove debris. Brain weight, the weight of the MFC pellet and the volume of 8M guanidine buffer are given examplary in the following table.
- ELISA Enzyme-Linked-lmmunosorbent-Assay
- the standard dilutions with final concentrations of 1000, 500, 250, 125, 62.5, 31.3 and 15.6 pg/ml were prepared in sample diluent with the same concentration guanidine as for the samples. This was performed in a 96-well polypropylene plate without protein binding capacity (Greiner bio-one, Frickenhausen, Germany).
- Samples, standards and blanks (50 ⁇ l) were added to the anti-A ⁇ -coated polystyrol plate (capture antibody selectively recognizes the C-terminal end of the antigen) in addition with a selective anti-A ⁇ -antibody conjugate (biotinylated detection antibody) and incubated overnight at 4°C in order to allow formation of the antibody-Amyloid-antibody-complex. The following day, a streptavidin-peroxidase conjugate was added, followed 30 minutes later by the addition of a TMB/peroxide mixture, resulting in the conversion of the substrate into a coloured product.
- a ⁇ (1 -42) monomer 0.1 % NH 4 OH
- a ⁇ (1-42) globulomer 0.1 % NH 4 OH
- a ⁇ (1-42) globulomer 0.1 % NH 4 OH
- Example 7 Analysis of cognitive performance by object recognition test in APP/L transgenic mice after passive immunization with anti-A ⁇ (20-42) globulomer antibodies
- mice overexpressing human APP with a point mutation were used.
- the point mutation refers to amino acid 717 (substitution of isoleucine for valine) and has been found in a London family where it leads to onset of AD before the beginning of the sixth decade of life (Mullan et al., Nature Genetics 2 (1992) 340-342).
- the transgenic mice, herein referred to as APP/L were created by and first described in Leuven (Moechars et al., J. Biol. Chem. 274 (1999) 6483-6492). Female APP/L mice were subjected to passive immunization at 3 months of age.
- mice received 250 ⁇ g of any of the monoclonal mouse antibodies 5F7, 10F11 or 7C6 in 100 ⁇ l of phosphate-buffered saline (PBS). Throughout the time course of the experiment the animals were kept under standard conditions in a reverted day/night cycle (14 hours of light beginning at 7 pm/10 hours of darkness). They tolerated passive immunization well, without any signs of adverse effects.
- PBS phosphate-buffered saline
- the ..recognition index expresses this relation (time for new object/total time).
- a mouse which does not remember the known object will consider it as equally interesting as the new object and spend an equal amount of time on exploring it, in other words, will show a recognition index of 50%.
- a mouse which remembers the known object will consider it as not interesting and therefore show a significantly higher recognition index.
- APP/L mice are known to be cognitively deficient at 4.5 months of age and exhibit a recognition index in the dimension of the random level, i.e. 50%. Results are shown in figure 6.
- Object recognition test in mice reports recognition of a known object in comparison to an unknown one, measured in terms of explorative behaviour during a 10 minute test phase.
- the recognition index is defined as the percentage of time which the mouse spends on exploring the unknown object relative to the time spent on exploring both objects.
- the known object was explored by the mouse during a 10 minute acquisition phase 2.5 hours before the test phase.
- APP/L mice are known to be cognitively deficient at 4.5 months of age and exhibit a recognition index in the dimension of the random level, i.e. 50%.
- Example 8 Dot-Blot profile of the selectivity of the anti-A ⁇ (20-42) globulomer antibodies.
- a ⁇ (20-42) globulomer The preparation of the A ⁇ (20-42) globulomer is described in example 1c.
- a ⁇ (1-42) (Bachem Inc. cat. no.: H-1368) were solved in 500 ⁇ l aqueous 0.1% NH 4 OH (Eppendorff tube) and the sample was stirred for 1 min at room temperature. 100 ⁇ l of this freshly prepared A ⁇ (1-42) solution were neutralized with 300 ⁇ l 20 mM NaH 2 PO 4 ; 140 mM
- sAPP ⁇ Supplied by Sigma (cat.no. S9564; 25 ⁇ g in 20 mM NaH 2 PO 4 ; 140 mM NaCI; pH 7.4).
- Detection reagent BM Blue POD Substrate, precipitating (Roche)
- BSA Bovine Serum Albumin
- Antibody solution II 1 :5000 dilution
- the dot blot was incubated with 30 ml 5% low fat milk in TBS for 1.5 h at RT.
- the blocking solution was discarded and the dot blot was incubated under shaking with 20 ml TTBS for 10 min at RT.
- Antibody solution I The washing buffer was discarded and the dot blot was incubated with antibody solution I for 2 h at RT
- the antibody solution Il was discarded and the dot blot was incubated under shaking with 20 ml TTBS for 10 min at RT.
- the washing solution was discarded and the dot blot was incubated under shaking with 20 ml TTBS for 10 min at RT.
- the washing solution was discarded and the dot blot was incubated under shaking with 20 ml TBS for 10 min at RT.
- the washing solution was discarded.
- the dot blot was developed with 10 ml BM Blue POD Substrate for 10 min. The development was stopped by intense washing of the dot blot with H 2 O. Quantitative evaluation was done using a densitometric analysis (GS800 densitometer (BioRad) and software package Quantity one, Version 4.5.0 (BioRad)) of the dot-intensity. Only dots were evaluted that had a relative density of greater than 20% of the relative density of the last optically unambiguously identified dot of the A ⁇ (20-42) globulomer. This threshold value was determined for every dot-blot independently. The calculated value indicates the relation between recognition of A ⁇ (20-42) globulomer and the respective A ⁇ form for the antibody given.
- the anti A ⁇ (20-42) globulomer selective mAbs can be divided in 3 classes with respect to the discrimination of A ⁇ (1-42) globulomer and A ⁇ ( 12-42) globulomer.
- the first class comprising the antibodies 6A2, 5F7 and 2F2 recognizes preferentially A ⁇ (20-42) globulomer and to some extent A ⁇ (1-42) globulomer (and also A ⁇ ( 12-42) globulomer).
- the second class comprising the antibodies 10F11 , 4D10 and 3B10 recognizes preferentially A ⁇ (20-42) globulomer and also recognizes A ⁇ ( 12-42) globulomer but to a lesser extent and do not significantly recognize A ⁇ (1-42) globulomer.
- the third class comprising the antibodies 7C6, 4B7, 7E5 and 10C1 rec- ognizes A ⁇ (20-42) globulomer but shows no significant recognition of the others. All three classes do not significantly recognize monomeric A ⁇ (1-42), monomeric A ⁇ (1-40), A ⁇ (1-42) fibrils or sAPP ⁇ .
- the selectivity profile of the anti-A ⁇ (20-42) globulomer antibodies shows that the significantly elevated recognition index in the passive immunization (in figure 6) must mainly be due to a selective recognition of truncated A ⁇ (20-42) globulomer and A ⁇ (12-42) globulomer and to a much lesser extent of A ⁇ (1 -42) globulomer and not monomeric A ⁇ ( 1-42), monomeric A ⁇ ( 1-40), A ⁇ (1-42) fibrils or sAPP ⁇ .
- Example 9 In situ analysis of the specific reaction of A ⁇ (20-42) selective antibodies to fibrillary A ⁇ peptide in the form of A ⁇ plaques in old TG2576 mice and A ⁇ amyloid in meningeal vessels.
- brain material of 19 month old TG2576 mice (Hsiao et al., 1996, Sci- ence; 274(5284), 99-102) or 9 month old APP/LxPS1 mice (description as above; ReMYND, Leuven, Belgium) or autopsy material of two Alzheimer's disease patients (RZ16 and RZ55; obtained from BrainNet, Kunststoff) was used.
- mice overexpress human APP with the so-called Swedish mutation (K670N/M671 L; Tg2576) or with the so-called London mutation (V717I) in addition with the human Presenilin 1 gene with the A264E mutation (APP/LxPS1 ) and formed ⁇ amyloid deposits in the brain parenchyma at about 7-11 months of age and ⁇ amyloid deposits in larger cerebral vessels at about 18 months of age (Tg2576).
- the animals were deeply anaesthetized and transcardially perfused with 0.1 M phosphate-buffered saline (PBS) to flush the blood. Then the brain was removed from the cranium and divided longitudinally.
- PBS phosphate-buffered saline
- One hemisphere of the brain was shock-frozen, the other fixated by immersion into 4% paraformaldehyde.
- the immersion-fixated hemisphere was cryoprotected by soaking in 30% sucrose in PBS and mounted on a freezing microtome. The entire forebrain was cut into 40 ⁇ m section which were collected in PBS and used for the subsequent staining procedure.
- the human brain material was an about 1 cm 3 deep-frozen block of the neocortex. A small part of the block was immersion-fixated in 4% paraformaldehyde and further treated like the mouse brain material.
- Antibody staining was performed by incubating the sections with a solution containing 0.07-7.0 ⁇ g/ml of the respective antibody in accordance with the following protocol:
- biotinylated donkey-anti-mouse antibody Jackson Immuno; 715-065- 150; diluted 1 :500 in TBST
- StreptABComplex DakoCytomation; K 0377
- plaque staining was additionally quantified by graphically excising 10 randomly selected plaques from the histological images using the ImagePro 5.0 image analysis system and determining their average greyscale value. Optical density values were calculated from the greyscale values by subtracting the mean background density of the stained material from the density of amyloid plaques (0% - no plaque staining above surrounding background, 100% - no transmission / maximal staining), and the differences between control and antibodies and between 6G1 and the antibodies selective for A ⁇ (20-42), respectively, were tested for statistical significance with ANOVA.
- Parenchymal A ⁇ deposits (amyloid plaques) were stained only with 6G1 and 6E10 but not with the globulomer selective antibodies (i.e. 5F7, 2F2, 6A2, 4D10, 10F11 , 3B10, 7C6,
- Parenchymal A ⁇ deposits were significantly more and at lower concentrations stained with 6G1 , 6E10 and 4G8 than with the globulomer selective antibodies (i.e. 5F7, 2F2, 6A2, 4D10, 10F1 1 , 3B10, 7C6, 7E5 and 10C1 ).
- Parenchymal A ⁇ deposits were stained only with 6G1 and 6E10 but not with the globulomer selective antibodies (i.e. 5F7, 2F2, 6A2, 4D10, 10F11 , 3B10, 7C6, 7E5 and 10C1 ).
- All globulomer selective antibodies i.e. 5F7, 2F2, 6A2, 4D10, 10F11 , 3B10, 7C6, 7E5 and 10C1
- Vascular A ⁇ deposits (arrows) were stained only with 6G1 and 6E10 but not with globulomer selective antibodies (i.e. 5F7, 2F2, 6A2, 4D10, 10F11 , 3B10, 7C6, 7E5 and 10C1 ). Binding of different antibodies at a concentration of 0.07-7.0 ⁇ g/ml in transversal section of the neocortices of transgenic APP/LxPS1 mice at 11 months of age:
- Parenchymal A ⁇ deposits were significantly more and at lower concentrations stained with 6G1 , 6E10 and 4G8 than with the globulomer selective antibodies (i.e. 5F7, 2F2, 6A2, 4D10, 10F11 , 3B10, 7C6, 7E5 and 10C1 ). All amyloid deposits had been verified by congophilic staining before (Congo Red; see Fig. 8A).
- Example 10 Anti-A ⁇ -antibody titer and dot-blot selectivity profile in plasma of TG2576 mice approximately one year after active immunization.
- a ⁇ (20-42) globulomer The preparation of the A ⁇ (20-42) globulomer is described in example 1c.
- a ⁇ (1 -42) fibrils 1 mg A ⁇ (1-42) (Bachem Inc. Catalog Nr.: H-1368) were solved in 500 ⁇ l aqueous 0.1%
- a ⁇ -standards Serial dilution of A ⁇ -antigens in 20 mM NaH 2 PO 4 , 140 mM NaCI, pH 7.4
- Nitrocellulose Trans-Blot Transfer medium, Pure Nitrocellulose Membrane (0.45 ⁇ m);
- Antibody solution II 1 :5000 dilution
- the dot blot is incubated with 30 ml 5% low fat milk in TBS for 1.5 h at RT.
- the blocking solution is discareded and the dot blot incubated under shaking with 20 ml TTBS for 10 min at RT.
- Antibody solution I The washing buffer is discarded and the dot blot incubated with antibody solution I overnight at RT
- the antibody solution I is discarded and the dot blot incubated under shaking with 20 ml TTBS for 10 min at RT.
- the washing solution is discarded and the dot blot incubated under shaking with 20 ml TTBS for 10 min at RT.
- the washing solution is discarded and the dot blot incubated under shaking with 20 ml TBS for 10 min at RT.
- Antibody solution II The washing buffer is discarded and the dot blot incubated with antibody solution Il for
- the antibody solution Il is discarded and the dot blot incubated under shaking with 20 ml TTBS for 10 min at RT.
- the washing solution is discarded and the dot blot incubated under shaking with 20 ml TTBS for 10 min at RT.
- the washing solution is discarded and the dot blot incubated under shaking with 20 ml TBS for 10 min at RT.
- Sera of different immunization groups a) A ⁇ (20-42) globulomers; b) A ⁇ (12-42) globulomers; c) A ⁇ (1-42) monomer, 0.1% NH 4 OH; d) vehicle control were tested against different A ⁇ forms in a dot blot for differing antibody profiles.
- a ⁇ (12-42) globulomer 5.
- the immunization with A ⁇ (20-42) globulomer or A ⁇ ( 12-42) globulomer exhibits even after approximately one year of the last immunization a high titer of antibodies.
- These antibodies are selective for the A ⁇ (20-42) globulomer in the case of the A ⁇ (20-42) globulomer immunization or A ⁇ (20-42) globulomer and A ⁇ ( 12-42) globulomer selective in the case of the A ⁇ ( 12-42) globu- lomer immunization.
- Example 11 Brain-levels of A ⁇ (20-42) globulomer epitopes in Alzheimer's disease patients.
- AD brain samples RZ 16; RZ 52 und RZ 55 (obtained from Brain Net, Kunststoff)
- Control sample RZ 92 (obtained from Brain Net, Kunststoff)
- 100 mg of AD brain sample are homogenized in 2.5 ml NaH 2 PO 4 , 140 mM NaCI, 0.05% Tween 20, 0.5% BSA (supplemented with 25 ⁇ l protease inhibitor solution) with 20 strokes in a glass potter. The suspension is sonified for 30 sec on ice, then incubated at 37 0 C for 16 h.
- the suspension is centrifuged at 100'0OO g and 8 0 C for one hour, then the supernatant is collected.
- the residue is dissolved in 5 mM NaH 2 PO 4 , 35 mM NaCI, pH 7.4 and homogenized with 10 strokes in a glass potter.
- 75 ⁇ l of 10% SDS and 125 ⁇ l of 0.16 mg/ml DTT are added and stirred for 20 minutes at ambient temperature.
- the sample was centrifuged for 10 minutes at 10'0OO g, and the supernatant is stored overnight at -20 0 C. Before use the supernatant is thawed and centrifuged for another 10 min at 10'0OO g.
- the individual binding antibodies 5F7, 7C6 and 10F1 1 are diluted to a final concentration of 0.7 ⁇ g/ml in coupling buffer.
- the blocking reagent is dissolved in 100 ml H 2 O and stored at -20 0 C in aliquots of 10 ml each.
- 3 ml of the blocking stock solution are diluted with 27 ml H 2 O for blocking one ELISA plate.
- a ⁇ (1-42) globulomer The preparation of the A ⁇ (1-42) globulomer is described in example 1a.
- the anti-A ⁇ pRAb stock solution is diluted to 0.05 ⁇ g/ml in PBST + 0,5 %BSA.
- the an- tibody solution is used immediately.
- Lyophilized anti-rabbit-POD conjugate is dissolved in 0.5 ml H 2 O and mixed with 500 ⁇ l glycerol.
- the antibody concentrate is then stored at -20 0 C in aliquots of 100 ⁇ l.
- the concentrate is diluted 1 :10'000 in PBST buffer. The antibody solution is used immediately.
- This ELISA is performed with each of the binding monoclonal antibodies 5F7, 7C6, 10F11.
- a sandwich ELISA was used to assess brain extracts for their truncated A ⁇ (20-42) globulomer epitope content.
- ELISAs with the respective antibodies against the A ⁇ (20-42) globulomer were used for calibration.
- Extraction of Alzheimer's disease brain tissue shows that the A ⁇ (20-42) globulomer epitope content is significantly elevated compared to a control patient. This shows that indeed the A ⁇ (20-42) globulomer epitope is a relevant A ⁇ -species in human Alzheimer's disease brain and not only relevant for Alzheimer's disease animal models. Antibodies directed against the A ⁇ (20-42) globulomer epitope therefore are highly desirable for the treatment of Alzheimer's disease.
- Monoclonal antibodies can be prepared using a wide variety of techniques known in the art including the use of hybridoma, recombinant, and phage display technologies, or a combination thereof.
- monoclonal antibodies can be produced using hybridoma techniques including those known in the art and taught, for example, in Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed. 1988); Hammerling, et al., in: Monoclonal Antibodies and T-CeII Hybridomas 563-681 (Elsevier, N. Y., 1981 ) (said references incorporated herein by reference in their entireties).
- the term “monoclonal antibody” as used herein is not limited to antibodies produced through hybridoma technology.
- the term “monoclonal antibody” refers to an antibody that is derived from a single clone, including any eukaryotic, prokaryotic, or phage clone, and not the method by which it is produced.
- mice Balb/c and A/J mice (6-8week old) were immunized subcutaneously with 50 ug of antigen in CFA. Animals were boosted every three weeks with 50 ug of antigen in ImmuneasyTM (Qiagen) for a total of three boosts. Four days prior to fusion, mice were boosted with 10 ug of antigen intravenously.
- Cell fusion and hybridoma screening Spleen cells from immunized animals were fused with SP2/0-Ag14 myeloma cells at a ratio of 5:1 using standard techniques. Seven to ten days post fusion, when macroscopic colonies were observed, SN were tested by ELISA for antibody to A ⁇ (20-42) globulomer. Cells from ELISA positive wells were scaled up and cloned by limiting dilution.
- Antibody isotype determination The isotype of the anti-A ⁇ (20-42) globulomer mAbs was de- termined using the Zymed EIA isotyping kit. Scale up and purification of monoclonal antibodies: Hybridomas were expanded into media containing 5% Low IgG. Fetal bovine serum (Hyclone). Supernatant was harvested and concentrated. mAb was purified using Protein A chromatography and dialyzed into PBS.
- Serum titers Ten mice were immunized with the A ⁇ (20-42) globulomer. All mice seroconverted with ELISA titers (1/2 Max OD 450 nm) of 1 :5000-10,000.
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US11299751B2 (en) | 2016-04-29 | 2022-04-12 | Voyager Therapeutics, Inc. | Compositions for the treatment of disease |
US11326182B2 (en) | 2016-04-29 | 2022-05-10 | Voyager Therapeutics, Inc. | Compositions for the treatment of disease |
PT3672631T (en) | 2017-08-22 | 2023-05-23 | Biogen Ma Inc | Pharmaceutical compositions containing anti-beta amyloid antibodies |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2004067561A1 (en) | 2003-01-31 | 2004-08-12 | Abbott Gmbh & Co. Kg | Amyloid-$g(b)(1-42) oligomers, derivatives thereof, antibodies for the same, method for production and use therof |
Family Cites Families (732)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE2500076C3 (en) | 1975-01-02 | 1982-11-18 | SCHURA Blutderivate GmbH & Co KG, 4150 Krefeld | Process for the production of intravenously tolerated gamma globulins |
US4634665A (en) | 1980-02-25 | 1987-01-06 | The Trustees Of Columbia University In The City Of New York | Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials |
US5179017A (en) | 1980-02-25 | 1993-01-12 | The Trustees Of Columbia University In The City Of New York | Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials |
US4399216A (en) | 1980-02-25 | 1983-08-16 | The Trustees Of Columbia University | Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials |
NO812612L (en) | 1980-08-06 | 1982-02-08 | Ferring Pharma Ltd | ENZYME inhibitor. |
WO1982001011A1 (en) | 1980-09-24 | 1982-04-01 | Corp Cetus | Diagnostic method and probe |
US4582788A (en) | 1982-01-22 | 1986-04-15 | Cetus Corporation | HLA typing method and cDNA probes used therein |
DE3381518D1 (en) | 1982-01-22 | 1990-06-07 | Cetus Corp | METHOD FOR CHARACTERIZING HLA AND THE CDNS TEST AGENTS USED IN IT. |
DE3378250D1 (en) | 1982-04-22 | 1988-11-24 | Ici Plc | Continuous release formulations |
US4510245A (en) | 1982-11-18 | 1985-04-09 | Chiron Corporation | Adenovirus promoter system |
GB8308235D0 (en) | 1983-03-25 | 1983-05-05 | Celltech Ltd | Polypeptides |
US4816567A (en) | 1983-04-08 | 1989-03-28 | Genentech, Inc. | Recombinant immunoglobin preparations |
US4526039A (en) | 1983-06-23 | 1985-07-02 | The United States Of America As Represented By The Secretary Of Transportation | Removable strain gauge fixture and method for measuring accumulated strain in a material |
US4683194A (en) | 1984-05-29 | 1987-07-28 | Cetus Corporation | Method for detection of polymorphic restriction sites and nucleic acid sequences |
US5807715A (en) | 1984-08-27 | 1998-09-15 | The Board Of Trustees Of The Leland Stanford Junior University | Methods and transformed mammalian lymphocyte cells for producing functional antigen-binding protein including chimeric immunoglobulin |
US5128326A (en) | 1984-12-06 | 1992-07-07 | Biomatrix, Inc. | Drug delivery systems based on hyaluronans derivatives thereof and their salts and methods of producing same |
US5168062A (en) | 1985-01-30 | 1992-12-01 | University Of Iowa Research Foundation | Transfer vectors and microorganisms containing human cytomegalovirus immediate-early promoter-regulatory DNA sequence |
US4683202A (en) | 1985-03-28 | 1987-07-28 | Cetus Corporation | Process for amplifying nucleic acid sequences |
US6492107B1 (en) | 1986-11-20 | 2002-12-10 | Stuart Kauffman | Process for obtaining DNA, RNA, peptides, polypeptides, or protein, by recombinant DNA technique |
DE229046T1 (en) | 1985-03-30 | 1987-12-17 | Marc Genf/Geneve Ballivet | METHOD FOR OBTAINING DNA, RNS, PEPTIDES, POLYPEPTIDES OR PROTEINS BY DNA RECOMBINANT METHOD. |
US4666829A (en) | 1985-05-15 | 1987-05-19 | University Of California | Polypeptide marker for Alzheimer's disease and its use for diagnosis |
US4980286A (en) | 1985-07-05 | 1990-12-25 | Whitehead Institute For Biomedical Research | In vivo introduction and expression of foreign genetic material in epithelial cells |
US5618920A (en) | 1985-11-01 | 1997-04-08 | Xoma Corporation | Modular assembly of antibody genes, antibodies prepared thereby and use |
US4968615A (en) | 1985-12-18 | 1990-11-06 | Ciba-Geigy Corporation | Deoxyribonucleic acid segment from a virus |
DE3600905A1 (en) | 1986-01-15 | 1987-07-16 | Ant Nachrichtentech | METHOD FOR DECODING BINARY SIGNALS AND VITERBI DECODERS AND APPLICATIONS |
CA1284931C (en) | 1986-03-13 | 1991-06-18 | Henry A. Erlich | Process for detecting specific nucleotide variations and genetic polymorphisms present in nucleic acids |
GB8607679D0 (en) | 1986-03-27 | 1986-04-30 | Winter G P | Recombinant dna product |
US5225539A (en) | 1986-03-27 | 1993-07-06 | Medical Research Council | Recombinant altered antibodies and methods of making altered antibodies |
US5107065A (en) | 1986-03-28 | 1992-04-21 | Calgene, Inc. | Anti-sense regulation of gene expression in plant cells |
CA1338457C (en) | 1986-08-22 | 1996-07-16 | Henry A. Erlich | Purified thermostable enzyme |
US5231170A (en) | 1986-08-27 | 1993-07-27 | Paul Averback | Antibodies to dense microspheres |
US5525339A (en) | 1986-08-27 | 1996-06-11 | Dms Pharmaceutical Inc. | Isolated components of dense microspheres derived from mammalian brain tissue and antibodies thereto |
US4946778A (en) | 1987-09-21 | 1990-08-07 | Genex Corporation | Single polypeptide chain binding molecules |
US5811310A (en) | 1986-09-30 | 1998-09-22 | Albert Einstein College Of Medicine Of Yeshiva Univ. | The Alz-50 monoclonal antibody and diagnostic assay for alzheimer's disease |
JPS63240797A (en) | 1986-10-14 | 1988-10-06 | Shin Etsu Chem Co Ltd | Monoclonal antibody, production thereof and diagnostic containing said antibody |
JPH0779702B2 (en) | 1986-11-17 | 1995-08-30 | サイオス ノバ インコーポレイテッド | Recombinant Alzheimer amyloid protein |
DE3702789A1 (en) | 1987-01-30 | 1988-08-18 | Bayer Ag | PROCUREMENT PROTEIN OF APC POLYPEPTIDE, FOR CODING DNA AND DIAGNOSTIC USE OF DNA AND PROTEIN |
DE3883899T3 (en) | 1987-03-18 | 1999-04-22 | Sb2, Inc., Danville, Calif. | CHANGED ANTIBODIES. |
JPS63245689A (en) | 1987-03-31 | 1988-10-12 | Suntory Ltd | Human amyloid-related protein monoclonal antibody |
US5258498A (en) | 1987-05-21 | 1993-11-02 | Creative Biomolecules, Inc. | Polypeptide linkers for production of biosynthetic proteins |
US4880078A (en) | 1987-06-29 | 1989-11-14 | Honda Giken Kogyo Kabushiki Kaisha | Exhaust muffler |
CA1340802C (en) | 1987-08-15 | 1999-10-26 | Yasuyuki Takahashi | Senile amyloid precursor protein and an antibody specific for the same |
US5004697A (en) | 1987-08-17 | 1991-04-02 | Univ. Of Ca | Cationized antibodies for delivery through the blood-brain barrier |
US5231000A (en) | 1987-10-08 | 1993-07-27 | The Mclean Hospital | Antibodies to A4 amyloid peptide |
CA1339014C (en) | 1987-10-08 | 1997-03-25 | Ronald E. Majocha | Antibodies to a4 amyloid peptide |
AU3056289A (en) | 1988-01-13 | 1989-08-11 | Mclean Hospital Corporation, The | Genetic constructs containing the alzheimer brain amyloid gene |
AU3204689A (en) | 1988-02-10 | 1989-09-06 | Children's Medical Center Corporation | Amyloid protein precursors, genetic probes, antibodies, and methods of use |
US5134062A (en) | 1988-03-22 | 1992-07-28 | Cornell Research Foundation, Inc. | Diagnosis of neuronal disorders and screening potential therapeutic agents therefor |
US5015570A (en) | 1988-05-13 | 1991-05-14 | Molecular Therapeutics, Inc. | Molecular diagnosis of Alzheimer Disease |
US6287793B1 (en) | 1988-08-19 | 2001-09-11 | Elan Pharmaceuticals, Inc. | Diagnostic methods for alzheimer's disease |
DE68927933T2 (en) | 1988-09-02 | 1997-08-14 | Dyax Corp | PRODUCTION AND SELECTION OF RECOMBINANT PROTEINS WITH DIFFERENT BINDING POINTS |
US5223409A (en) | 1988-09-02 | 1993-06-29 | Protein Engineering Corp. | Directed evolution of novel binding proteins |
WO1990005144A1 (en) | 1988-11-11 | 1990-05-17 | Medical Research Council | Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors |
US5530101A (en) | 1988-12-28 | 1996-06-25 | Protein Design Labs, Inc. | Humanized immunoglobulins |
US20030229208A1 (en) | 1988-12-28 | 2003-12-11 | Protein Design Labs, Inc. | Humanized immunoglobulins |
US5231020A (en) | 1989-03-30 | 1993-07-27 | Dna Plant Technology Corporation | Genetic engineering of novel plant phenotypes |
US5262332A (en) | 1989-04-05 | 1993-11-16 | Brigham And Women's Hospital | Diagnostic method for Alzheimer's disease: examination of non-neural tissue |
AU5525090A (en) | 1989-04-14 | 1990-11-16 | Research Foundation For Mental Hygiene, Inc. | Monoclonal antibody to amyloid peptide |
CA2016841C (en) | 1989-05-16 | 1999-09-21 | William D. Huse | A method for producing polymers having a preselected activity |
CA2016842A1 (en) | 1989-05-16 | 1990-11-16 | Richard A. Lerner | Method for tapping the immunological repertoire |
AU652539B2 (en) | 1989-05-16 | 1994-09-01 | Medical Research Council | Co-expression of heteromeric receptors |
US5234814A (en) | 1989-06-01 | 1993-08-10 | Du Pont Merck Pharmaceutical Company | Diagnostic assay for alzheimer's disease |
EP0415801A1 (en) | 1989-07-05 | 1991-03-06 | N.V. Innogenetics S.A. | Monoclonal antibodies against activated microglial cells |
EP0411974A1 (en) | 1989-07-05 | 1991-02-06 | N.V. Innogenetics S.A. | Monoclonal antibodies to a neurofibrillary tangle antigen |
AU6430190A (en) | 1989-10-10 | 1991-05-16 | Pitman-Moore, Inc. | Sustained release composition for macromolecular proteins |
AU642932B2 (en) | 1989-11-06 | 1993-11-04 | Alkermes Controlled Therapeutics, Inc. | Protein microspheres and methods of using them |
GB8928874D0 (en) | 1989-12-21 | 1990-02-28 | Celltech Ltd | Humanised antibodies |
US5780225A (en) | 1990-01-12 | 1998-07-14 | Stratagene | Method for generating libaries of antibody genes comprising amplification of diverse antibody DNAs and methods for using these libraries for the production of diverse antigen combining molecules |
WO1991010737A1 (en) | 1990-01-11 | 1991-07-25 | Molecular Affinities Corporation | Production of antibodies using gene libraries |
DE69133566T2 (en) | 1990-01-12 | 2007-12-06 | Amgen Fremont Inc. | Formation of xenogenic antibodies |
US6075181A (en) | 1990-01-12 | 2000-06-13 | Abgenix, Inc. | Human antibodies derived from immunized xenomice |
AU7121191A (en) | 1990-02-26 | 1991-10-10 | Albert Einstein College Of Medicine Of Yeshiva University | Diagnostic assay for alzheimer's disease |
US5753624A (en) | 1990-04-27 | 1998-05-19 | Milkhaus Laboratory, Inc. | Materials and methods for treatment of plaquing disease |
US5427908A (en) | 1990-05-01 | 1995-06-27 | Affymax Technologies N.V. | Recombinant library screening methods |
GB9015198D0 (en) | 1990-07-10 | 1990-08-29 | Brien Caroline J O | Binding substance |
JPH06508511A (en) | 1990-07-10 | 1994-09-29 | ケンブリッジ アンティボディー テクノロジー リミティド | Method for producing specific binding pair members |
DE4022247A1 (en) | 1990-07-12 | 1992-01-16 | Hoechst Ag | METHOD FOR EXTRACTIVELY SEPARATING 2-ALKYLTHIO-5-PHENYLPYRIMIDINES FROM THEIR REACTION MIXTURES |
EP0542810A1 (en) | 1990-08-02 | 1993-05-26 | B.R. Centre Limited | Methods for the production of proteins with a desired function |
EP0814159B1 (en) | 1990-08-29 | 2005-07-27 | GenPharm International, Inc. | Transgenic mice capable of producing heterologous antibodies |
US5625126A (en) | 1990-08-29 | 1997-04-29 | Genpharm International, Inc. | Transgenic non-human animals for producing heterologous antibodies |
US5770429A (en) | 1990-08-29 | 1998-06-23 | Genpharm International, Inc. | Transgenic non-human animals capable of producing heterologous antibodies |
US5814318A (en) | 1990-08-29 | 1998-09-29 | Genpharm International Inc. | Transgenic non-human animals for producing heterologous antibodies |
US6255458B1 (en) | 1990-08-29 | 2001-07-03 | Genpharm International | High affinity human antibodies and human antibodies against digoxin |
US5545806A (en) | 1990-08-29 | 1996-08-13 | Genpharm International, Inc. | Ransgenic non-human animals for producing heterologous antibodies |
US5633425A (en) | 1990-08-29 | 1997-05-27 | Genpharm International, Inc. | Transgenic non-human animals capable of producing heterologous antibodies |
US5661016A (en) | 1990-08-29 | 1997-08-26 | Genpharm International Inc. | Transgenic non-human animals capable of producing heterologous antibodies of various isotypes |
US5877397A (en) | 1990-08-29 | 1999-03-02 | Genpharm International Inc. | Transgenic non-human animals capable of producing heterologous antibodies of various isotypes |
US5698426A (en) | 1990-09-28 | 1997-12-16 | Ixsys, Incorporated | Surface expression libraries of heteromeric receptors |
DE69129154T2 (en) | 1990-12-03 | 1998-08-20 | Genentech, Inc., South San Francisco, Calif. | METHOD FOR ENRICHING PROTEIN VARIANTS WITH CHANGED BINDING PROPERTIES |
JPH04252195A (en) | 1991-01-29 | 1992-09-08 | Asahi Chem Ind Co Ltd | Monoclonal antibody and hybridoma |
EP0575485A1 (en) | 1991-03-01 | 1993-12-29 | Dyax Corp. | Process for the development of binding mini-proteins |
JP3672306B2 (en) | 1991-04-10 | 2005-07-20 | ザ スクリップス リサーチ インスティテュート | Heterodimeric receptor library using phagemids |
JPH04320694A (en) | 1991-04-22 | 1992-11-11 | Ono Pharmaceut Co Ltd | Monoclonal antibody against human gliacyte growth inhibiting factor |
ES2181673T3 (en) | 1991-05-01 | 2003-03-01 | Jackson H M Found Military Med | PROCESS OF TREATMENT OF INFECTIOUS RESPIRATORY DISEASES. |
JPH04342883A (en) | 1991-05-17 | 1992-11-30 | Sanden Corp | Variable delivery swash plate type compressor |
DE69233482T2 (en) | 1991-05-17 | 2006-01-12 | Merck & Co., Inc. | Method for reducing the immunogenicity of antibody variable domains |
CA2110799A1 (en) | 1991-06-14 | 1992-12-23 | Arnold H. Horwitz | Microbially-produced antibody fragments and their conjugates |
DE4122599C2 (en) | 1991-07-08 | 1993-11-11 | Deutsches Krebsforsch | Phagemid for screening antibodies |
ES2136092T3 (en) | 1991-09-23 | 1999-11-16 | Medical Res Council | PROCEDURES FOR THE PRODUCTION OF HUMANIZED ANTIBODIES. |
WO1993008302A1 (en) | 1991-10-25 | 1993-04-29 | Itt Automotive Europe Gmbh | Monoclonal antibodies directed against the microtubule-associated protein tau |
JP3277211B2 (en) | 1991-11-12 | 2002-04-22 | プラナ・バイオテクノロジー・リミテッド | Test and treatment methods for Alzheimer's disease |
ATE408012T1 (en) | 1991-12-02 | 2008-09-15 | Medical Res Council | PRODUCTION OF AUTOANTIBODIES ON PHAGE SURFACES BASED ON ANTIBODIES SEGMENT LIBRARIES |
US20020009443A1 (en) | 1991-12-02 | 2002-01-24 | Vanitha Ramakrishman | Inhibitory immunoglobulin polypeptides to human pdgf beta receptor |
ATE438716T1 (en) | 1991-12-06 | 2009-08-15 | Max Planck Gesellschaft | USE OF PROTEIN KINASES TO DIAGNOSE AND TREAT ALZHEIMER'S DISEASE |
US7408027B1 (en) | 1991-12-06 | 2008-08-05 | Max-Planck-Gesellschaft Zur Forderung Der Wissenchaften | Tools for the diagnosis and treatment of Alzheimer's disease |
DE69233204T2 (en) | 1991-12-13 | 2004-07-15 | Xoma Corp., Berkeley | METHOD AND MATERIALS FOR THE PRODUCTION OF MODIFIED VARIABLE ANTIBODY DOMAINS AND THEIR THERAPEUTIC USE |
US20010029293A1 (en) | 1992-01-27 | 2001-10-11 | Icos Corporation | Icam-related protein |
US5538845A (en) | 1992-02-05 | 1996-07-23 | Athena Neurosciences, Inc. | Beta-amyloid peptide production inhibitors and methods for their identification |
US5714350A (en) | 1992-03-09 | 1998-02-03 | Protein Design Labs, Inc. | Increasing antibody affinity by altering glycosylation in the immunoglobulin variable region |
US5912015A (en) | 1992-03-12 | 1999-06-15 | Alkermes Controlled Therapeutics, Inc. | Modulated release from biocompatible polymers |
US5733743A (en) | 1992-03-24 | 1998-03-31 | Cambridge Antibody Technology Limited | Methods for producing members of specific binding pairs |
US5912120A (en) | 1992-04-09 | 1999-06-15 | The United States Of America As Represented By The Department Of Health And Human Services, | Cloning, expression and diagnosis of human cytochrome P450 2C19: the principal determinant of s-mephenytoin metabolism |
US5604102A (en) | 1992-04-15 | 1997-02-18 | Athena Neurosciences, Inc. | Methods of screening for β-amyloid peptide production inhibitors |
US5441870A (en) | 1992-04-15 | 1995-08-15 | Athena Neurosciences, Inc. | Methods for monitoring cellular processing of β-amyloid precursor protein |
US5455169A (en) | 1992-06-04 | 1995-10-03 | Alzheimer's Institute Of America, Inc. | Nucleic acids for diagnosing and modeling Alzheimer's disease |
US6610493B1 (en) | 1993-06-17 | 2003-08-26 | Brigham And Women's Hospital | Screening compounds for the ability to alter the production of amyloid-β peptide |
US5766846A (en) | 1992-07-10 | 1998-06-16 | Athena Neurosciences | Methods of screening for compounds which inhibit soluble β-amyloid peptide production |
US5837672A (en) | 1992-07-10 | 1998-11-17 | Athena Neurosciences, Inc. | Methods and compositions for the detection of soluble β-amyloid peptide |
EP0652950B1 (en) | 1992-07-24 | 2007-12-19 | Amgen Fremont Inc. | Generation of xenogeneic antibodies |
US5639641A (en) | 1992-09-09 | 1997-06-17 | Immunogen Inc. | Resurfacing of rodent antibodies |
TW258789B (en) | 1992-09-28 | 1995-10-01 | Rohm & Haas | |
US5605811A (en) | 1992-10-26 | 1997-02-25 | Athena Neurosciences, Inc. | Methods and compositions for monitoring cellular processing of beta-amyloid precursor protein |
US5891623A (en) | 1992-11-09 | 1999-04-06 | Consorzio Per Le Biotecnologie | Diagnosis and treatment of AIDS onset |
ZA938243B (en) | 1992-11-12 | 1995-05-04 | Hybritech Inc | Altered affinity polypeptides of metal chelate binding antibodies |
US6010913A (en) | 1992-12-14 | 2000-01-04 | N.V. Innogenetics S.A. | Isolated human tau peptide |
EP0683234B2 (en) | 1993-01-25 | 2007-06-06 | Takeda Chemical Industries, Ltd. | Antibody against beta-amyloid or their derivative and use thereof |
US5955317A (en) | 1993-01-25 | 1999-09-21 | Takeda Chemical Industries, Ltd. | Antibodies to β-amyloids or their derivatives and use thereof |
US5934272A (en) | 1993-01-29 | 1999-08-10 | Aradigm Corporation | Device and method of creating aerosolized mist of respiratory drug |
CA2115900A1 (en) | 1993-02-22 | 1994-08-23 | Gerald W. Becker | Pharmaceutical screens and antibodies |
US5840294A (en) | 1993-03-29 | 1998-11-24 | Queen's University At Kingston | Method for treating amyloidosis |
WO1995007707A1 (en) | 1993-09-14 | 1995-03-23 | Cytel Corporation | Alteration of immune response using pan dr-binding peptides |
WO1995011311A1 (en) | 1993-10-20 | 1995-04-27 | Duke University | METHOD OF BINDING MATERIAL TO THE β-AMYLOID PEPTIDE |
US5565352A (en) | 1993-11-24 | 1996-10-15 | Arch Development Corporation | Deubiquitinating enzyme: compositions and methods |
WO1995015982A2 (en) | 1993-12-08 | 1995-06-15 | Genzyme Corporation | Process for generating specific antibodies |
AU1397495A (en) | 1993-12-13 | 1995-07-03 | Uab Research Foundation, The | Diagnostic tests and reagents for detecting risk of alzheimer's disease and stroke |
US6432404B1 (en) | 1993-12-23 | 2002-08-13 | Icos Corporation | Methods of inhibiting locomotor damage following spinal cord injury with α D-specific antibodies |
US6251395B1 (en) | 1993-12-23 | 2001-06-26 | W. Michael Gallatin | Methods of inhibiting inflammation at the site of a central nervous system injury with alphaD-specific antibodies |
US6620915B2 (en) | 1993-12-23 | 2003-09-16 | Icos Corporation | Monoclonal antibodies specific for integrin α-d subunit |
JPH07209295A (en) | 1994-01-10 | 1995-08-11 | Nikken Chem Co Ltd | Antibody, enzyme-labeled antigen and method and kit for enzyme immunoassay |
JPH07209296A (en) | 1994-01-10 | 1995-08-11 | Nikken Chem Co Ltd | Antibody, enzyme-labeled antigen and method and kit for enzyme immunoassay |
US5877399A (en) | 1994-01-27 | 1999-03-02 | Johns Hopkins University | Transgenic mice expressing APP-Swedish mutation develop progressive neurologic disease |
DE69534347T2 (en) | 1994-01-31 | 2006-05-24 | Trustees Of Boston University, Boston | Libraries of polyclonal antibodies |
JPH07238096A (en) | 1994-02-25 | 1995-09-12 | S R L:Kk | Antipolyubiquitin-monoclonal antibody and method for measuring polyubiquitin |
US7473423B2 (en) | 1994-04-29 | 2009-01-06 | Mayo Foundation For Medical Education And Research | Human IgM antibodies, and diagnostic and therapeutic uses thereof particularly in the central nervous system |
JPH07309900A (en) | 1994-05-20 | 1995-11-28 | Idemitsu Kosan Co Ltd | Anti-human procathepsin b monoclonal antibody, hybridoma producting the same and assay of human procathepsin b or human cathepsin b using the same |
US5516637A (en) | 1994-06-10 | 1996-05-14 | Dade International Inc. | Method involving display of protein binding pairs on the surface of bacterial pili and bacteriophage |
US6018021A (en) | 1994-10-19 | 2000-01-25 | The Research Foundation Of State University Of New York | Human transaldolase: an autoantigen with a function in metabolism |
US6114133A (en) | 1994-11-14 | 2000-09-05 | Elan Pharmaceuticals, Inc. | Methods for aiding in the diagnosis of Alzheimer's disease by measuring amyloid-β peptide (x-≧41) |
ATE252894T1 (en) | 1995-01-05 | 2003-11-15 | Univ Michigan | SURFACE-MODIFIED NANOPARTICLES AND METHODS FOR THEIR PRODUCTION AND USE |
US5786180A (en) | 1995-02-14 | 1998-07-28 | Bayer Corporation | Monoclonal antibody 369.2B specific for β A4 peptide |
JP3663228B2 (en) | 1995-03-14 | 2005-06-22 | 株式会社医学生物学研究所 | Antibodies against neuropsin |
US6130364A (en) | 1995-03-29 | 2000-10-10 | Abgenix, Inc. | Production of antibodies using Cre-mediated site-specific recombination |
US6091001A (en) | 1995-03-29 | 2000-07-18 | Abgenix, Inc. | Production of antibodies using Cre-mediated site-specific recombination |
US6019968A (en) | 1995-04-14 | 2000-02-01 | Inhale Therapeutic Systems, Inc. | Dispersible antibody compositions and methods for their preparation and use |
US5705330A (en) | 1995-04-14 | 1998-01-06 | Abbott Laboratories | Chemiluminescent immunoassay for antibody detection |
EP0826034A4 (en) | 1995-04-21 | 2002-06-19 | Cell Genesys Inc | Generation of large genomic dna deletions |
ATE390933T1 (en) | 1995-04-27 | 2008-04-15 | Amgen Fremont Inc | HUMAN ANTIBODIES AGAINST IL-8 DERIVED FROM IMMUNIZED XENOMICES |
AU2466895A (en) | 1995-04-28 | 1996-11-18 | Abgenix, Inc. | Human antibodies derived from immunized xenomice |
US5712158A (en) | 1995-06-06 | 1998-01-27 | Warner-Lambert Company | Cell line for the rapid expression of functional calcium channels |
US6717031B2 (en) | 1995-06-07 | 2004-04-06 | Kate Dora Games | Method for selecting a transgenic mouse model of alzheimer's disease |
AU6255096A (en) | 1995-06-07 | 1996-12-30 | Mount Sinai School Of Medicine Of The City University Of New York, The | Pegylated modified proteins |
JP3767755B2 (en) | 1995-06-23 | 2006-04-19 | エーザイ株式会社 | Anti-JC virus antibody |
US6127977A (en) | 1996-11-08 | 2000-10-03 | Cohen; Nathan | Microstrip patch antenna with fractal structure |
CA2229043C (en) | 1995-08-18 | 2016-06-07 | Morphosys Gesellschaft Fur Proteinoptimierung Mbh | Protein/(poly)peptide libraries |
JP2000507912A (en) | 1995-08-31 | 2000-06-27 | アルカームズ コントロールド セラピューティックス,インコーポレイテッド | Sustained release composition of active agent |
JP3495738B2 (en) | 1995-09-14 | 2004-02-09 | ザ レジェンツ オブ ザ ユニバーシティー オブ カリフォルニア | Natural PrPsc-specific antibody |
FR2740454B1 (en) | 1995-10-26 | 1997-11-21 | Rhone Poulenc Rorer Sa | PEPTIDES CAPABLE OF INHIBITING APP ENDOCYTOSIS AND CORRESPONDING NUCLEOTIDE SEQUENCES |
AUPN649395A0 (en) | 1995-11-10 | 1995-12-07 | Ramsay Health Care Pty Ltd | A method for diagnosing alzheimer's disease |
FR2741881B1 (en) | 1995-12-01 | 1999-07-30 | Centre Nat Rech Scient | NOVEL PURINE DERIVATIVES HAVING IN PARTICULAR ANTI-PROLIFERATIVE PRORIETES AND THEIR BIOLOGICAL APPLICATIONS |
KR19980702156A (en) | 1995-12-12 | 1998-07-15 | 히라따 다다시 | New DNA, New Polypeptides and New Antibodies |
JPH09178743A (en) | 1995-12-27 | 1997-07-11 | Oriental Yeast Co Ltd | Determinationof soluble app |
ATE219247T1 (en) | 1996-01-19 | 2002-06-15 | Res Corp Technologies Inc | METHODS FOR DETECTING ALZHEIMER'S DISEASE |
JP2978435B2 (en) | 1996-01-24 | 1999-11-15 | チッソ株式会社 | Method for producing acryloxypropyl silane |
EP0948536B1 (en) | 1996-01-30 | 2007-06-13 | The Scripps Research Institute | A g protein-coupled receptor with an enlarged extracellular domain |
BRPI9715219B8 (en) | 1996-02-09 | 2015-07-07 | Abbvie Biotechnology Ltd | Recombinant expression vector, and prokaryotic host cell. |
ES2395214T3 (en) | 1996-03-04 | 2013-02-11 | The Penn State Research Foundation | Materials and methods to increase cellular internalization |
US5714352A (en) | 1996-03-20 | 1998-02-03 | Xenotech Incorporated | Directed switch-mediated DNA recombination |
US5882644A (en) | 1996-03-22 | 1999-03-16 | Protein Design Labs, Inc. | Monoclonal antibodies specific for the platelet derived growth factor β receptor and methods of use thereof |
US5874064A (en) | 1996-05-24 | 1999-02-23 | Massachusetts Institute Of Technology | Aerodynamically light particles for pulmonary drug delivery |
US5985309A (en) | 1996-05-24 | 1999-11-16 | Massachusetts Institute Of Technology | Preparation of particles for inhalation |
US5855913A (en) | 1997-01-16 | 1999-01-05 | Massachusetts Instite Of Technology | Particles incorporating surfactants for pulmonary drug delivery |
US6300065B1 (en) | 1996-05-31 | 2001-10-09 | Board Of Trustees Of The University Of Illinois | Yeast cell surface display of proteins and uses thereof |
US6699658B1 (en) | 1996-05-31 | 2004-03-02 | Board Of Trustees Of The University Of Illinois | Yeast cell surface display of proteins and uses thereof |
WO1997046678A1 (en) | 1996-06-06 | 1997-12-11 | Bayer Corporation | Nucleic acids and polypeptides related to presenilin |
WO1998006403A1 (en) * | 1996-08-13 | 1998-02-19 | P.N. Gerolymatos S.A. | Use of the chelating agent clioquinol for the manufacture of a pharmaceutical composition for the treatment of alzheimer's disease |
WO1999009150A1 (en) | 1996-08-15 | 1999-02-25 | Bayer Corporation | Method of introducing modifications into a gene |
CA2183901A1 (en) | 1996-08-22 | 1998-02-23 | Johanna E. Bergmann | Targets for therapy and diagnosis of alzheimer's disease and down syndrome in humans |
US7521531B2 (en) | 1996-08-28 | 2009-04-21 | Immunomedics, Inc. | Methods for the purification of stable radioiodine conjugates |
JPH1075781A (en) | 1996-08-30 | 1998-03-24 | Sumitomo Electric Ind Ltd | Human proteasome subunit p58 protein and specific antibody against the same |
EP0834561A1 (en) | 1996-09-27 | 1998-04-08 | Rijksuniversiteit te Leiden | A gene related to migraine in man |
US5916771A (en) | 1996-10-11 | 1999-06-29 | Abgenix, Inc. | Production of a multimeric protein by cell fusion method |
WO1998022120A1 (en) | 1996-11-19 | 1998-05-28 | The Wistar Institute Of Anatomy & Biology | Diagnostic and therapeutic reagents for alzheimer's disease |
AU5702298A (en) | 1996-12-03 | 1998-06-29 | Abgenix, Inc. | Transgenic mammals having human Ig loci including plural VH and VK regions nd antibodies produced therefrom |
US5919687A (en) | 1996-12-24 | 1999-07-06 | John Hopkins University | Recombinant N-SMases and nucleic acids encoding same |
WO1998031346A1 (en) | 1997-01-16 | 1998-07-23 | Massachusetts Institute Of Technology | Preparation of particles for inhalation |
CN1238366C (en) | 1997-01-21 | 2006-01-25 | 综合医院公司 | Selection of proteins using RNA-protein fusions |
JPH10210982A (en) | 1997-01-31 | 1998-08-11 | Fuji Yakuhin Kogyo Kk | New protein |
US20030068316A1 (en) | 1997-02-05 | 2003-04-10 | Klein William L. | Anti-ADDL antibodies and uses thereof |
US6218506B1 (en) | 1997-02-05 | 2001-04-17 | Northwestern University | Amyloid β protein (globular assembly and uses thereof) |
US6413940B1 (en) | 1997-02-07 | 2002-07-02 | Nymox Corporation | Pharmaceutically active agents that impede the formation of amyloid by impeding the genesis of DMS |
US7226730B1 (en) | 1997-02-26 | 2007-06-05 | The General Hospital Corporation | Transgenic animals and cell lines for screening drugs effective for the treatment or prevention of Alzheimer's Disease |
US7045531B1 (en) | 1997-03-11 | 2006-05-16 | The General Hospital Corporation | Composition comprising a metal chelator and a method of treating amyloidosis by administering the metal chelator |
AU6513298A (en) | 1997-03-17 | 1998-10-12 | British Technology Group Limited | Therapeutic compositions |
WO1998047343A2 (en) | 1997-04-04 | 1998-10-29 | Biosite Diagnostics, Inc. | Antibodies or binding protein libraries displayed on phage, cells, or other replicatable genetic packages |
US20020086847A1 (en) | 1997-04-09 | 2002-07-04 | Mindset Biopharmaceuticals (Usa) | Recombinant antibodies specific for beta-amyloid ends, DNA encoding and methods of use thereof |
US8173127B2 (en) | 1997-04-09 | 2012-05-08 | Intellect Neurosciences, Inc. | Specific antibodies to amyloid beta peptide, pharmaceutical compositions and methods of use thereof |
WO1998049286A2 (en) | 1997-05-01 | 1998-11-05 | Board Of Regents, The University Of Texas System | Directed evolution of enzymes and antibodies |
US6235883B1 (en) | 1997-05-05 | 2001-05-22 | Abgenix, Inc. | Human monoclonal antibodies to epidermal growth factor receptor |
DE69838249T3 (en) | 1997-05-15 | 2012-01-19 | Genentech, Inc. | ANTI-APO-2 ANTIBODIES |
US6689359B1 (en) | 1997-07-21 | 2004-02-10 | Arpi Matossian-Rogers | Ligands, including antibodies, showing reactivity against endocrine cells |
IT1293511B1 (en) | 1997-07-30 | 1999-03-01 | Gentili Ist Spa | MONOCLONAL CATALYTIC ANTIBODIES WITH PROTEASIC ACTIVITY FOR THE SELECTIVE LYSIS OF THE PROTEIN COMPONENT OF PLATES AND RELATED AGGREGATES |
ES2281135T3 (en) | 1997-08-14 | 2007-09-16 | Institut Pasteur | HYBRID PROTEINS OF A TETANIC TOXOID THAT MIGRATE BACK AND TRANSINAPTICALLY TO THE CNS. |
US7923216B2 (en) | 1997-08-14 | 2011-04-12 | Institut Pasteur | In vivo modulation of neuronal transport |
JP2001513576A (en) | 1997-08-26 | 2001-09-04 | グリアテック インコーポレイテッド | Process for inhibiting complement activation via the collateral pathway |
US6666935B1 (en) | 1997-09-09 | 2003-12-23 | The Regents Of The University Of California | Sol-gel manufactured energetic materials |
FR2768346B1 (en) | 1997-09-15 | 2002-04-19 | Fond Jean Dausset Ceph | COMPOUND FOR INHIBITION OF PRESENILINE 1 FOR THE PREPARATION OF A MEDICAMENT AND DIAGNOSTIC AGENT |
US20040185039A1 (en) | 2002-08-30 | 2004-09-23 | Heinz Kohler | Therapeutic applications of noncovalent dimerizing antibodies |
US5989463A (en) | 1997-09-24 | 1999-11-23 | Alkermes Controlled Therapeutics, Inc. | Methods for fabricating polymer-based controlled release devices |
SE512663C2 (en) | 1997-10-23 | 2000-04-17 | Biogram Ab | Active substance encapsulation process in a biodegradable polymer |
WO1999022024A1 (en) | 1997-10-24 | 1999-05-06 | Cornell Research Foundation, Inc. | Detection of neurodegenerative diseases |
US6761888B1 (en) | 2000-05-26 | 2004-07-13 | Neuralab Limited | Passive immunization treatment of Alzheimer's disease |
US6750324B1 (en) | 1997-12-02 | 2004-06-15 | Neuralab Limited | Humanized and chimeric N-terminal amyloid beta-antibodies |
US6710226B1 (en) | 1997-12-02 | 2004-03-23 | Neuralab Limited | Transgenic mouse assay to determine the effect of Aβ antibodies and Aβ Fragments on alzheimer's disease characteristics |
US6787523B1 (en) | 1997-12-02 | 2004-09-07 | Neuralab Limited | Prevention and treatment of amyloidogenic disease |
US7964192B1 (en) | 1997-12-02 | 2011-06-21 | Janssen Alzheimer Immunotherapy | Prevention and treatment of amyloidgenic disease |
US7179892B2 (en) | 2000-12-06 | 2007-02-20 | Neuralab Limited | Humanized antibodies that recognize beta amyloid peptide |
US6913745B1 (en) | 1997-12-02 | 2005-07-05 | Neuralab Limited | Passive immunization of Alzheimer's disease |
US6905686B1 (en) | 1997-12-02 | 2005-06-14 | Neuralab Limited | Active immunization for treatment of alzheimer's disease |
TWI239847B (en) | 1997-12-02 | 2005-09-21 | Elan Pharm Inc | N-terminal fragment of Abeta peptide and an adjuvant for preventing and treating amyloidogenic disease |
ES2253839T3 (en) | 1997-12-03 | 2006-06-01 | Neuralab, Ltd. | SUPPRESSION OF CHANGES RELATED TO AMILOID BETA IN THE DISEASE OF ALZHEIMER. |
AU1507499A (en) | 1997-12-15 | 1999-07-05 | Kyowa Hakko Kogyo Co. Ltd. | Human nap1 protein |
US6730778B2 (en) | 1997-12-19 | 2004-05-04 | Pharmacia And Upjohn Company | Human sel-10 polypeptides and polynucleotides that encode them |
US6436989B1 (en) | 1997-12-24 | 2002-08-20 | Vertex Pharmaceuticals, Incorporated | Prodrugs of aspartyl protease inhibitors |
US7189703B2 (en) | 1998-01-09 | 2007-03-13 | Intracell, Llc | Treatment and diagnosis of alzheimer's disease |
NZ550199A (en) | 1998-02-11 | 2008-07-31 | Bellus Health Int Ltd | Treatment of inflammation, abeta-induced cell toxicity, neuronal cell death or neuronal cell loss in subjects suffering from alzheimer's disease |
AU2978899A (en) | 1998-03-03 | 1999-09-20 | Abgenix, Inc. | Cd147 binding molecules as therapeutics |
US20050112543A1 (en) | 1998-03-11 | 2005-05-26 | The General Hospital Corporation | Method of screening for drugs useful in treating Alzheimer's disease |
US6323218B1 (en) | 1998-03-11 | 2001-11-27 | The General Hospital Corporation | Agents for use in the treatment of Alzheimer's disease |
US20050059591A1 (en) | 1998-04-07 | 2005-03-17 | Neuralab Limited | Prevention and treatment of amyloidogenic disease |
US20050059802A1 (en) | 1998-04-07 | 2005-03-17 | Neuralab Ltd | Prevention and treatment of amyloidogenic disease |
US20020029391A1 (en) | 1998-04-15 | 2002-03-07 | Claude Geoffrey Davis | Epitope-driven human antibody production and gene expression profiling |
US6913756B1 (en) | 1998-04-29 | 2005-07-05 | The Uab Research Foundation | Monoclonal antibodies specific for anthrax and peptides derived from the antibodies thereof |
NO314086B1 (en) | 1998-05-08 | 2003-01-27 | Gemvax As | Peptides and pharmaceutical compositions containing them, nucleic acid sequences encoding such peptides, plasmids and virus vectors encompassing such DNA sequences and their use for the preparation of pharmaceutical preparations for |
WO1999058157A1 (en) | 1998-05-08 | 1999-11-18 | Cramer Donald V | Method for inhibiting antibody-mediated rejection of xenogeneic tissues |
MXPA00011213A (en) | 1998-05-15 | 2003-04-22 | Neurochem Inc | Use of amyloid inhibitors for modulating neuronal cell death. |
US20030147882A1 (en) | 1998-05-21 | 2003-08-07 | Alan Solomon | Methods for amyloid removal using anti-amyloid antibodies |
ATE255888T1 (en) | 1998-06-01 | 2003-12-15 | Ortho Mcneil Pharm Inc | TETRAHYDRONAPHTALENE COMPOUNDS AND THEIR USE FOR THE TREATMENT OF NEURODEGENERATIVE DISEASES |
JP2000050885A (en) | 1998-06-02 | 2000-02-22 | Fuji Chemical Industries Ltd | Antibody against cathepsin and its utilization |
ATE238768T1 (en) | 1998-06-24 | 2003-05-15 | Advanced Inhalation Res Inc | LARGE POROUS PARTICLES EXPECTED FROM AN INHALER |
WO2000001807A1 (en) | 1998-07-02 | 2000-01-13 | Fuso Pharmaceutical Industries, Ltd. | Serine protease-specific monoclonal antibody and utilization thereof |
CA2341029A1 (en) | 1998-08-17 | 2000-02-24 | Abgenix, Inc. | Generation of modified molecules with increased serum half-lives |
US20030187191A1 (en) | 1998-09-01 | 2003-10-02 | Genentech, Inc. | Secreted and transmembrane polypeptides and nucleic acids encoding the same |
ATE346923T1 (en) | 1998-09-17 | 2006-12-15 | Otsuka Pharma Co Ltd | LY6H-GEN |
US7083950B2 (en) | 1998-09-25 | 2006-08-01 | Regeneron Pharmaceuticals, Inc. | High affinity fusion proteins and therapeutic and diagnostic methods for use |
AU5999199A (en) | 1998-09-29 | 2000-04-17 | Kyowa Hakko Kogyo Co. Ltd. | Novel antibodies, drugs containing these antibodies and methods for screening compounds by using these antibodies |
US6660843B1 (en) | 1998-10-23 | 2003-12-09 | Amgen Inc. | Modified peptides as therapeutic agents |
CA2353795A1 (en) | 1998-12-02 | 2000-06-08 | The Trustees Of The University Of Pennsylvania | Methods and compositions for determining lipid peroxidation levels in oxidant stress syndromes and diseases |
EP1731913A3 (en) | 1998-12-02 | 2007-01-17 | The Trustees Of The University Of Pennsylvania | Methods and compositions for determining lipid peroxidation levels in oxidant stress syndromes and diseases |
DK1140969T3 (en) | 1998-12-14 | 2007-09-17 | Univ Miami | Connective tissue growth factor fragments and methods and uses thereof |
PT2112166T (en) | 1998-12-23 | 2019-01-30 | Pfizer | Human monoclonal antibodies to ctla-4 |
DE19902550A1 (en) | 1999-01-22 | 2000-07-27 | Memorec Medical Molecular Rese | A novel aspartate protease, coding sequence, inhibitors and antibodies, useful for treatment and diagnosis of Alzheimer's disease |
US20030185826A1 (en) | 1999-02-24 | 2003-10-02 | Tobinick Edward L. | Cytokine antagonists for the treatment of localized disorders |
CA2796140A1 (en) | 1999-03-25 | 2000-09-28 | Jochen Salfeld | Human antibodies that bind human il-12 and methods for producing |
FR2791263B1 (en) | 1999-03-26 | 2001-04-27 | Halina Zofia Malina | DRUG PREPARATIONS BASED ON AN IMMUNE RESPONSE AGAINST ACCUMULATION OF XANTHURENIC ACID MODIFIED PROTEINS |
CA2704600C (en) | 1999-04-09 | 2016-10-25 | Kyowa Hakko Kirin Co., Ltd. | A method for producing antibodies with increased adcc activity |
US7527787B2 (en) | 2005-10-19 | 2009-05-05 | Ibc Pharmaceuticals, Inc. | Multivalent immunoglobulin-based bioactive assemblies |
US6787637B1 (en) | 1999-05-28 | 2004-09-07 | Neuralab Limited | N-Terminal amyloid-β antibodies |
PE20010212A1 (en) | 1999-06-01 | 2001-02-22 | Neuralab Ltd | COMPOSITIONS OF THE A-BETA PEPTIDE AND PROCESSES TO PRODUCE THEM |
UA81216C2 (en) | 1999-06-01 | 2007-12-25 | Prevention and treatment of amyloid disease | |
US20030027158A1 (en) | 1999-06-03 | 2003-02-06 | Curagen Corporation | Novel nucleic acid sequences encoding human breast tumor-associated protein 47-like polypeptides |
JP2000354487A (en) | 1999-06-15 | 2000-12-26 | Takashi Muramatsu | Midkine receptor |
US20020002270A1 (en) | 1999-06-16 | 2002-01-03 | Raymond P. Zinkowski | Purified antigen for alzheimer's disease, and methods of obtaining and using same |
WO2000077178A1 (en) | 1999-06-16 | 2000-12-21 | Boston Biomedical Research Institute | IMMUNOLOGICAL CONTROL OF β-AMYLOID LEVELS IN VIVO |
JP4796725B2 (en) * | 1999-08-04 | 2011-10-19 | ユニバーシティ オブ サザン カリフォルニア | Amyloid β protein (spherical assembly and use thereof) |
US7605238B2 (en) | 1999-08-24 | 2009-10-20 | Medarex, Inc. | Human CTLA-4 antibodies and their uses |
KR100942863B1 (en) | 1999-08-24 | 2010-02-17 | 메다렉스, 인코포레이티드 | Human ctla-4 antibodies and their uses |
DE60016242T2 (en) | 1999-09-01 | 2005-12-01 | Evotec Neurosciences Gmbh | METHOD FOR DIAGNOSIS OR FORECAST OF AGE-RELATED MACULATE GENERATION |
ATE461996T1 (en) | 1999-09-03 | 2010-04-15 | Univ Ramot | COMPOUNDS, COMPOSITIONS AND METHODS FOR THE TREATMENT OR PREVENTION OF ALZHEIMER'S DISEASE |
US20040013647A1 (en) | 1999-09-03 | 2004-01-22 | Ramot At Tel-Aviv University Ltd. | Methods and compositions for treating a plaque-forming disease |
US6294171B2 (en) | 1999-09-14 | 2001-09-25 | Milkhaus Laboratory, Inc. | Methods for treating disease states comprising administration of low levels of antibodies |
JP2001122900A (en) | 1999-10-21 | 2001-05-08 | Yasukazu Tanuma | ANTI-DNASE gamma ANTIBODY AND ITS PREPARATION AND USE |
EP1230269A2 (en) | 1999-11-03 | 2002-08-14 | Maxygen, Inc. | Antibody diversity generation |
US20070135337A2 (en) | 1999-11-29 | 2007-06-14 | Neurochem (International) Limited | Vaccine for the Prevention and Treatment of Alzheimer's and Amyloid Related Diseases |
CA2388559A1 (en) | 1999-11-29 | 2001-06-07 | Neurochem Inc. | Vaccine for the prevention and treatment of alzheimer's and amyloid related diseases |
US20020094335A1 (en) | 1999-11-29 | 2002-07-18 | Robert Chalifour | Vaccine for the prevention and treatment of alzheimer's and amyloid related diseases |
EP1752472A3 (en) | 1999-12-08 | 2007-04-25 | Intellect Neurosciences, Inc. | Chimeric amyloid beta peptides |
EP1237930B1 (en) | 1999-12-08 | 2006-11-08 | Intellect Neurosciences, Inc. | Chimeric amyloid beta peptides |
JP2001231578A (en) | 1999-12-09 | 2001-08-28 | Kyowa Hakko Kogyo Co Ltd | Protein belonging to il-1 family |
IL150374A0 (en) | 1999-12-23 | 2002-12-01 | Neurochem Inc | Compounds and methods for modulating cerebral amyloid angiopathy |
US20030077757A1 (en) | 2000-01-11 | 2003-04-24 | Andrews William H. | Method of treating aging-related disorders |
WO2001054318A1 (en) | 2000-01-17 | 2001-07-26 | Corning O.T.I. S.P.A. | Attenuator integrated with modulator and transmitting module for wdm system using the same |
US6806254B2 (en) | 2000-02-03 | 2004-10-19 | Nuvelo, Inc. | Methods and materials relating to alpha-2-macroglobulin-like polypeptides and polynucleotides |
US20020182660A1 (en) | 2000-02-18 | 2002-12-05 | Fong Kei-Lai L. | N- and C-terminus specific immunoassays for full length beta-amyloid peptide-Abeta(1-40), Abeta(1-39), Abeta(1-40), Abeta(1-42) , and Abeta(1-43) |
DE60114157T2 (en) | 2000-02-21 | 2006-06-29 | Pharmexa A/S | METHOD OF HERBEGULATING AMYLOID |
KR20080017471A (en) | 2000-02-21 | 2008-02-26 | 파멕사 에이/에스 | Novel method for down-regulation of amyloid |
CZ2008595A3 (en) | 2000-02-24 | 2017-05-03 | Washington University | A medication for the prevention or treatment of preclinical or clinical Alzheimer's disease |
EP1130032A1 (en) | 2000-02-28 | 2001-09-05 | Gesellschaft für biotechnologische Forschung mbH (GBF) | Single-chain antibodies recognizing the human vascular endothelial growth factor receptor-2 (VEGFR-2/KDR) |
GB0006398D0 (en) | 2000-03-16 | 2000-05-03 | Novartis Ag | Organic compounds |
WO2001071351A1 (en) | 2000-03-22 | 2001-09-27 | The General Hospital Corporation | Method for treatment of neurodegenerative diseases |
WO2001075165A2 (en) | 2000-03-30 | 2001-10-11 | Elan Pharmaceuticals, Inc. | Screening markers and methods for neurodegenerative disorders |
US20020164668A1 (en) | 2000-04-03 | 2002-11-07 | Durham L. Kathryn | Nucleic acid molecules, polypeptides and uses therefor, including diagnosis and treatment of alzheimer's disease |
US7371365B2 (en) | 2000-04-04 | 2008-05-13 | Mayo Foundation For Medical Education And Research | Methods for detecting parenchymal plaques in vivo |
AUPQ712000A0 (en) | 2000-04-27 | 2000-05-18 | Medvet Science Pty. Ltd. | Antigenic peptide fragments of VapA protein, and uses thereof |
US20080009467A1 (en) | 2000-05-01 | 2008-01-10 | Accera, Inc. | Combinations of medium chain triglycerides and therapeutic agents for the treatment and prevention of alzheimers disease and other diseases resulting from reduced neuronal metabolism |
JP2003533187A (en) | 2000-05-03 | 2003-11-11 | アムジエン・インコーポレーテツド | Modified peptide containing Fc domain as therapeutic agent |
WO2001090182A2 (en) | 2000-05-22 | 2001-11-29 | New York University | Synthetic immunogenic but non-amyloidogenic peptides homologous to amyloid beta for induction of an immune response to amyloid beta and amyloid deposits |
DK1292680T3 (en) | 2000-06-22 | 2010-03-08 | Genentech Inc | Agonist-anti-TrkC monoclonal antibodies |
JP4252195B2 (en) | 2000-06-27 | 2009-04-08 | 新日本無線株式会社 | High frequency line converter |
CN1450908A (en) | 2000-06-28 | 2003-10-22 | 普拉纳生物技术有限公司 | Neurotoxic oligomers |
US7449308B2 (en) | 2000-06-28 | 2008-11-11 | Glycofi, Inc. | Combinatorial DNA library for producing modified N-glycans in lower eukaryotes |
EP1297172B1 (en) | 2000-06-28 | 2005-11-09 | Glycofi, Inc. | Methods for producing modified glycoproteins |
US6686449B2 (en) | 2000-06-30 | 2004-02-03 | Pharmacia & Upjohn Company | Mutant presenilin 1 polypeptides |
WO2002002769A1 (en) | 2000-07-06 | 2002-01-10 | Vlaams Interuniversitair Instituut Voor Biotechnologie Vzw | A novel app mutation associated with an unusual alzheimer's disease pathology |
EP1309341A2 (en) | 2000-07-07 | 2003-05-14 | Lars Lannfelt | Prevention and treatment of alzheimer's disease |
AU2007200047B2 (en) | 2000-07-07 | 2009-11-26 | Bioarctic Neuroscience Ab | Prevention and treatment of Alzheimer's disease |
US6524819B1 (en) | 2000-07-11 | 2003-02-25 | Incyte Genomics, Inc. | Down syndrome critical region 1-like proteins |
US20020009445A1 (en) | 2000-07-12 | 2002-01-24 | Yansheng Du | Human beta-amyloid antibody and use thereof for treatment of alzheimer's disease |
EP1172378A1 (en) | 2000-07-12 | 2002-01-16 | Richard Dr. Dodel | Human beta-amyloid antibody and use thereof for treatment of alzheimer's disease |
JP2002040023A (en) | 2000-07-28 | 2002-02-06 | Mitsubishi Chemicals Corp | Detection method of alzheimer's disease |
US20070009931A1 (en) | 2000-08-04 | 2007-01-11 | Kirsch Wolff M | Negative correlation between IRP-2 and Transferrin receptor expression as a diagnostic of Alzheimer's disease |
US7067133B2 (en) | 2000-09-06 | 2006-06-27 | Aventis Pharma S.A. | Methods and compositions for diseases associated with amyloidosis |
GB0024446D0 (en) | 2000-10-05 | 2000-11-22 | Glaxo Group Ltd | Protein |
IT1319277B1 (en) | 2000-10-24 | 2003-09-26 | Chiesi Farma Spa | MELTING PROTEINS USEFUL FOR ALZHEIMER'S MILK IMMUNIZATION TREATMENT. |
US6962793B2 (en) | 2000-10-27 | 2005-11-08 | Mount Sinai Hospital | Methods for detecting Alzheimers disease |
EP1363939A2 (en) | 2000-11-01 | 2003-11-26 | Insight Biotechnology Limited | Peptides for use in the treatment of alzheimer's disease |
WO2002094870A2 (en) | 2000-11-02 | 2002-11-28 | Curagen Corporation | Proteins and nucleic acids encoding same |
DE10055703A1 (en) | 2000-11-02 | 2002-05-08 | Peter F Pascoe | Ex vivo blood lavage with oligomeric carbohydrates for removal of pathogens, hapten-containing antibody aggregates and amyloids, useful in treatment of e.g. multiple sclerosis and Alzheimer's disease |
EP1341548A4 (en) | 2000-11-03 | 2006-06-14 | Massachusetts Inst Technology | METHODS FOR IDENTIFYING TREATMENTS FOR NEUROTOXICITY IN ALZHEIMER'S DISEASE CAUSED BY $g(b)-AMYLOID PEPTIDES |
US20060062786A1 (en) | 2000-11-08 | 2006-03-23 | Human Genome Sciences, Inc. | Antibodies that immunospecifically bind to TRAIL receptors |
DK1346041T3 (en) | 2000-11-27 | 2007-07-02 | Praecis Pharm Inc | Therapeutic agents and methods for their use in the treatment of an amyloidogenic disease |
TWI255272B (en) | 2000-12-06 | 2006-05-21 | Guriq Basi | Humanized antibodies that recognize beta amyloid peptide |
GB0100110D0 (en) | 2001-01-04 | 2001-02-14 | Univ Leeds | Modulation of calcium channel activity |
DE10101430B4 (en) | 2001-01-13 | 2008-10-02 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Soluble cyclic analogues for the modulation of amyloidogenesis |
US20020132758A1 (en) | 2001-01-18 | 2002-09-19 | Shell John W. | Method for identifying compounds to treat medical pathologies associated with molecular crystallization |
US7320793B2 (en) | 2001-01-19 | 2008-01-22 | Cytos Biotechnology Ag | Molecular antigen array |
AT500379B8 (en) | 2001-02-02 | 2009-08-15 | Axon Neuroscience | TAU PROTEINS |
US7175988B2 (en) | 2001-02-09 | 2007-02-13 | Human Genome Sciences, Inc. | Human G-protein Chemokine Receptor (CCR5) HDGNR10 |
US20040191264A1 (en) | 2001-02-19 | 2004-09-30 | Nielsen Klaus Gregorius | Synthetic vaccine agents |
JP4659236B2 (en) | 2001-03-01 | 2011-03-30 | 大塚製薬株式会社 | Method for measuring cytotoxic activity |
US6815175B2 (en) | 2001-03-16 | 2004-11-09 | Cornell Research Foundation, Inc. | Anti-amyloid peptide antibody based diagnosis and treatment of a neurological disease or disorder |
FR2822238B1 (en) | 2001-03-16 | 2003-08-29 | Gemac | METHOD AND KIT FOR MONITORING NEURODEGENERATIVE DISEASES |
DE10117281A1 (en) | 2001-04-06 | 2002-10-24 | Inst Molekulare Biotechnologie | Peptide for the diagnosis and therapy of Alzheimer's dementia |
US20050266501A1 (en) | 2001-04-10 | 2005-12-01 | Shigeo Ota | Reagent for detecting risk factor of alzheimer's disease, detection kit therefor and method of detecting risk factor of alzheimer's disease using the same |
CA2443770A1 (en) | 2001-04-23 | 2002-10-31 | Curagen Corporation | Proteins and nucleic acids encoding same |
US6451547B1 (en) | 2001-04-25 | 2002-09-17 | Syn X Pharma | Process for differential diagnosis of Alzheimer's dementia and device therefor |
DE60229051D1 (en) | 2001-04-30 | 2008-11-06 | Lilly Co Eli | HUMANIZED ANTIBODIES |
DE60230736D1 (en) | 2001-04-30 | 2009-02-26 | Lilly Co Eli | HUMANIZED ANTIBODIES WHICH RECOGNIZE THE BETA AMYLOID PEPTIDE &x9; |
US7196163B2 (en) | 2001-05-22 | 2007-03-27 | Merk & Co., Inc. | Assays using amyloid precursor proteins with modified β-secretase cleavage sites to monitor β-secretase activity |
DE60229846D1 (en) | 2001-05-22 | 2008-12-24 | Merck & Co Inc | BETA SECRETASE SUBSTRATE AND ITS USE |
US6906169B2 (en) | 2001-05-25 | 2005-06-14 | United Biomedical, Inc. | Immunogenic peptide composition comprising measles virus Fprotein Thelper cell epitope (MUFThl-16) and N-terminus of β-amyloid peptide |
ES2228697T3 (en) | 2001-06-12 | 2005-04-16 | Wiltfang, Jens | MONOCLONAL ANTIBODY, MAB 1E8, WHICH IS SPECIFIC FOR THE FIRST TWO AMINO ACIDS OF BETA-AMILOID PEPTIDES AND ITS USE IN THE BETA-AMILOID AND / OR SAPP ALFA PEPTIDES DETECTION. |
US7595378B2 (en) | 2001-06-13 | 2009-09-29 | Genmab A/S | Human monoclonal antibodies to epidermal growth factor receptor (EGFR) |
CA2450285C (en) | 2001-06-13 | 2016-08-02 | Genmab A/S | Human monoclonal antibodies to epidermal growth factor receptor (egfr) |
US7094884B2 (en) | 2001-06-22 | 2006-08-22 | Roche Diagnostics Corporation | Soluble complexes of amylod β peptide and peptidyl prolyl isomerase chaperone and methods of making and using them |
WO2003000714A2 (en) | 2001-06-22 | 2003-01-03 | Panacea Pharmaceuticals, Inc. | Compositions and methods for preventing protein aggregation in neurodegenerative diseases |
CN1396183A (en) | 2001-07-13 | 2003-02-12 | 张小如 | Human fusion antibody for reducing cerebral amyloid fibers associated with senile dementia |
AU2002326412A1 (en) | 2001-07-20 | 2003-03-03 | Oregon Health And Science University | Novel human nucleic acids encoding a pantothenate kinase and methods of use |
EP1420032B2 (en) | 2001-08-03 | 2015-12-16 | Medical & Biological Laboratories Co., Ltd. | Antibody recognizing gm1 ganglioside-bound amyloid beta-protein and dna encoding the antibody |
JP4320694B2 (en) | 2001-08-08 | 2009-08-26 | 株式会社オーク製作所 | Multiple exposure drawing apparatus and multiple exposure drawing method |
WO2003014329A2 (en) | 2001-08-10 | 2003-02-20 | University Of South Florida | Modulation of angiogenesis by a-beta peptides |
EP1519740A4 (en) | 2001-08-17 | 2005-11-09 | Lilly Co Eli | Rapid improvement of cognition in conditions related to a-beta |
CA2457145C (en) | 2001-08-17 | 2010-12-21 | Washington University | Assay method for alzheimer's disease |
JP2005500389A (en) | 2001-08-17 | 2005-01-06 | イーライ・リリー・アンド・カンパニー | Use of antibodies with high affinity for soluble Aβ to treat Aβ-related conditions and diseases |
PT1944040E (en) | 2001-08-17 | 2012-10-31 | Univ Washington | Assay method for alzheimer`s disease |
WO2003016466A2 (en) | 2001-08-17 | 2003-02-27 | Eli Lilly And Company | ANTI-Aβ ANTIBODIES |
MY144532A (en) | 2001-08-20 | 2011-09-30 | Lundbeck & Co As H | Novel method for down-regulation of amyloid |
US20030082191A1 (en) | 2001-08-29 | 2003-05-01 | Poduslo Joseph F. | Treatment for central nervous system disorders |
PT1438400E (en) | 2001-10-01 | 2009-09-10 | Dyax Corp | Multi-chain eukaryotic display vectors and uses thereof |
JP2003284574A (en) | 2001-10-03 | 2003-10-07 | Pfizer Prod Inc | Nucleic acid molecule, polypeptide and use therefor including diagnosis and treatment of alzheimer's disease |
JP2005508338A (en) | 2001-10-04 | 2005-03-31 | プロテイン セラピューティクス、インク. | Use of gamma globulin to treat immune diseases |
GB0124317D0 (en) | 2001-10-10 | 2001-11-28 | Celltech R&D Ltd | Biological products |
MXPA04003798A (en) | 2001-10-25 | 2004-07-30 | Genentech Inc | Glycoprotein compositions. |
CA2504349A1 (en) | 2001-11-02 | 2003-05-15 | Diagenics International Corporation | Monoclonal antibodies specific for beta-amyloid |
EP1572894B1 (en) | 2001-11-21 | 2016-04-13 | New York University | Synthetic immunogenic but non-deposit-forming polypeptides and peptides homologous to amyloid beta, prion protein, amylin, alpha synuclein, or polyglutamine repeats for induction of an immune response thereto |
US7026129B2 (en) | 2001-11-23 | 2006-04-11 | Syn X Pharma, Inc. | IG lambda biopolymer markers predictive of Alzheimers disease |
US7179606B2 (en) | 2001-11-23 | 2007-02-20 | Syn X Pharma, Inc. | IG heavy chain, IG kappa, IG lambda biopolymer markers predictive of Alzheimer's disease |
DE10158180A1 (en) | 2001-11-28 | 2003-09-11 | Biovision Ag | Method for the detection of Alzheimer's disease and to differentiate Alzheimer's disease from other dementia diseases, associated peptides and their use |
CA2468878A1 (en) | 2001-11-30 | 2003-06-05 | Crucell Holland B.V. | Antigen presenting cell targeting conjugate, an antigen presenting cell contacted with such conjugate, their use for vaccination or as medicament, and methods for their productionor generation |
IL162319A0 (en) | 2001-12-04 | 2005-11-20 | Univ Ben Gurion | Amphiphilic compounds and vesicles/liposomes for organ-specific drug targeting |
US20050227941A1 (en) | 2001-12-17 | 2005-10-13 | Karen Duff | Sequestration of ass in the periphery in the absence of immunomodulating agent as a therapeutic approach for the treatment or prevention of beta-amyloid related diseases |
EP1975179A1 (en) | 2001-12-26 | 2008-10-01 | Araclon Biotech, S.L. | Polyclonal antibodies, preparation method thereof and use of same |
US7781396B2 (en) | 2002-01-31 | 2010-08-24 | Tel Aviv University Future Technology Development L.P. | Peptides directed for diagnosis and treatment of amyloid-associated disease |
US20050153381A1 (en) | 2002-02-14 | 2005-07-14 | Marusich Michael F. | Immunocapture of mitochondrial protein complexes |
AR038568A1 (en) | 2002-02-20 | 2005-01-19 | Hoffmann La Roche | ANTI-A BETA ANTIBODIES AND ITS USE |
WO2003074004A2 (en) | 2002-03-01 | 2003-09-12 | Szu-Yi Chou | Method of producing antigens |
DE10208812A1 (en) | 2002-03-01 | 2003-09-11 | Basf Plant Science Gmbh | Process for the production of unsaturated fatty acids |
KR20040088572A (en) | 2002-03-01 | 2004-10-16 | 이뮤노메딕스, 인코오포레이티드 | Bispecific antibody point mutations for enhancing rate of clearance |
EP2292247A3 (en) | 2002-03-05 | 2011-10-05 | Ramot at Tel Aviv University Ltd. | Immunizing composition and method for inducing an immune response against the beta-secretase cleavage site of amyloid precursor protein |
MY139983A (en) | 2002-03-12 | 2009-11-30 | Janssen Alzheimer Immunotherap | Humanized antibodies that recognize beta amyloid peptide |
AU2003210060B2 (en) | 2002-03-22 | 2010-02-25 | Aprogen, Inc. | Humanized antibody and process for preparing same |
US20030190689A1 (en) | 2002-04-05 | 2003-10-09 | Cell Signaling Technology,Inc. | Molecular profiling of disease and therapeutic response using phospho-specific antibodies |
US7122374B1 (en) | 2002-04-09 | 2006-10-17 | Takaomi Saido | Amyloid beta-protein 3(pE)-42 antibodies and uses thereof |
WO2003086310A2 (en) | 2002-04-12 | 2003-10-23 | Ramot At Tel Aviv University Ltd. | Prevention of brain inflammation as a result of induced autoimmune response |
WO2003089460A1 (en) | 2002-04-19 | 2003-10-30 | The Governing Council Of The University Of Toronto | Immunological methods and compositions for the treatment of alzheimer's disease |
AU2003224120A1 (en) | 2002-04-24 | 2003-11-10 | Evotec Neurosciences Gmbh | Diagnostic and therapeutic use of ensadin-0477 gene and protein for neurodegenerative diseases |
ATE419871T1 (en) | 2002-04-25 | 2009-01-15 | Lilly Co Eli | METHOD FOR TREATING ANXIETY DISORDERS IN ELDERLY PERSONS |
US20060257420A1 (en) | 2002-04-26 | 2006-11-16 | Cel-Sci Corporation | Methods of preparation and composition of peptide constructs useful for treatment of autoimmune and transplant related host versus graft conditions |
DE10221052A1 (en) | 2002-05-10 | 2003-12-04 | Transmit Technologietransfer | Active substances for therapy, diagnostics and prophylaxis of diseases in which abnormal protein structures occur |
WO2003100419A1 (en) | 2002-05-27 | 2003-12-04 | Bioceros B.V. | Methods for using the cd163 pathway for modulating an immune response |
AU2003241131A1 (en) | 2002-06-14 | 2003-12-31 | Brainsgate Ltd. | Methods and systems for management of alzheimer's disease |
EP1514118A2 (en) | 2002-06-20 | 2005-03-16 | EVOTEC Neurosciences GmbH | Diagnostic and therapeutic use of ras-gtpase-activating sh3-domain-binding protein 2 (g3bp2) for neurodegenerative diseases |
AU2003260301A1 (en) | 2002-06-27 | 2004-01-19 | Evotec Neurosciences Gmbh | Diagnostic and therapeutic use of ensadin-0581 gene and protein for neurodegenerative diseases |
CN1678634A (en) | 2002-06-28 | 2005-10-05 | 多曼蒂斯有限公司 | Immunoglobulin single variable antigen combination area and its opposite constituent |
CN100577803C (en) | 2002-07-12 | 2010-01-06 | 阿克松神经科学研究和发展股份有限公司 | Express the transgenic animal of Alzheimer's tau protein |
AU2003256578A1 (en) | 2002-07-17 | 2004-02-02 | Intellect Neurosciences, Inc. | Peptides and methods of screening immunogenic peptide vaccines against alzheimer's disease |
JP4558485B2 (en) | 2002-07-18 | 2010-10-06 | 正康 大河内 | Novel polypeptide derived from Notch and biomarker and reagent using the same |
BR0312792A (en) | 2002-07-19 | 2005-05-03 | Cytos Biotechnology Ag | Vaccine compositions containing beta1-6 amyloid antigenic series |
KR20150043568A (en) | 2002-07-19 | 2015-04-22 | 애브비 바이오테크놀로지 리미티드 | Treatment of TNFα related disorders |
US7250551B2 (en) | 2002-07-24 | 2007-07-31 | President And Fellows Of Harvard College | Transgenic mice expressing inducible human p25 |
WO2004013172A2 (en) | 2002-07-24 | 2004-02-12 | Innogenetics N.V. | Fragments of beta-amyloid as targets for vaccination against alzheimer disease |
SE0202880D0 (en) | 2002-07-26 | 2002-09-30 | Wieslab Ab | Complement system deficiency assay |
WO2004011943A1 (en) | 2002-07-30 | 2004-02-05 | The Regents Of The University Of California | Method of diagnosing alzheimer's disease |
US20040138296A1 (en) | 2002-08-12 | 2004-07-15 | Pharmacia Corporation | Amyloid immunization and Cox-2 inhibitors for the treatment of alzheimer's disease |
US7252953B2 (en) | 2002-08-13 | 2007-08-07 | United States Of America Department Of Veterans Affairs | Method of detecting and preventing Alzheimer's disease, particularly at prodromal and early stages |
ES2352668T3 (en) | 2002-08-14 | 2011-02-22 | Mitsubishi Chemical Medience Corporation | SPECIFIC ANTIBODY FOR A TAU PROTEIN OF THE CENTRAL NERVOUS SYSTEM. |
AU2003259965A1 (en) | 2002-08-20 | 2004-03-11 | Neurogenetics, Inc. | Methods and compositions for modulating amyloid beta |
WO2004019045A2 (en) | 2002-08-23 | 2004-03-04 | Proteosys Ag | Method for the diagnosis of alzheimer disease |
WO2004024090A2 (en) | 2002-09-12 | 2004-03-25 | The Regents Of The University Of California | Immunogens and corresponding antibodies specific for high molecular weight aggregation intermediates common to amyloids formed from proteins of differing sequence |
US7238488B2 (en) | 2002-09-13 | 2007-07-03 | Grace Maresh | Therapeutic and diagnostic applications of perlecan domain I splice variants |
US20070010657A1 (en) | 2002-09-13 | 2007-01-11 | Rainer Klocke | Cytoplasmic dynein heavy chain 1 genes, expression products, non-human animal model uses in human neurological diseases |
CA2499599A1 (en) | 2002-09-17 | 2004-04-01 | New York University | Methods of treating age associated memory impairment (aami), mild cognitive impairment (mci), and dementias with cell cycle inhibitors |
JP4242128B2 (en) | 2002-09-18 | 2009-03-18 | 武田薬品工業株式会社 | Screening method for cerebral amyloidosis preventive and therapeutic drug |
WO2004029629A1 (en) | 2002-09-27 | 2004-04-08 | Janssen Pharmaceutica N.V. | N-11 truncated amyloid-beta nomoclonal antibodies, compositions, methods and uses |
US7541440B2 (en) | 2002-09-30 | 2009-06-02 | Immunomedics, Inc. | Chimeric, human and humanized anti-granulocyte antibodies and methods of use |
US20070213512A1 (en) | 2002-10-01 | 2007-09-13 | Krafft Grant A | Amyloid beta peptide analogs and assemblies thereof |
JP2006508072A (en) | 2002-10-01 | 2006-03-09 | ノースウエスタン ユニバーシティ | Amyloid beta-derived diffusible ligands (ADDLs), ADDL substitutes, ADDL-binding molecules, and uses thereof |
WO2004031241A1 (en) | 2002-10-04 | 2004-04-15 | Techno Network Shikoku Co., Ltd. | Monoclonal antibody against subtilisin-like proprotein convertase pace4 and utilization thereof |
CA2501831A1 (en) | 2002-10-09 | 2004-04-22 | Activx Biosciences, Inc. | Activity-based probes, and methods of their preparation and use |
EP1633786A4 (en) | 2002-10-09 | 2007-07-25 | Rinat Neuroscience Corp | Methods of treating alzheimer s disease using antibodies directed against amyloid beta peptide and compositions thereof |
WO2004038411A2 (en) | 2002-10-24 | 2004-05-06 | Evotec Neurosciences Gmbh | Diagnostic and therapeutic use of ensadin-0289 gene and protein for neurodegenerative diseases |
US20080014194A1 (en) | 2003-10-31 | 2008-01-17 | Elan Pharmaceuticals, Inc. | Prevention and Treatment of Synucleinopathic and Amyloidogenic Disease |
TW200509968A (en) | 2002-11-01 | 2005-03-16 | Elan Pharm Inc | Prevention and treatment of synucleinopathic disease |
FR2846667B1 (en) | 2002-11-06 | 2004-12-31 | Pasteur Institut | VARIABLE FRAGMENTS OF SINGLE-CHAIN CAMELIDE ANTIBODIES DIRECTED AGAINST BETA-AMYLOID PEPTIDE 1-42 AND THEIR APPLICATIONS FOR DIAGNOSIS AND TREATMENT OF NEUROAGREGATIVE DISEASES |
AU2003295411A1 (en) | 2002-11-07 | 2004-06-03 | Celltech R & D | Human monoclonal antibodies to heparanase |
AU2003294290A1 (en) | 2002-11-15 | 2004-06-15 | Morehouse School Of Medicine | Anti-chemokine and associated receptor antibodies and uses for inhibition of inflammation. |
US20040162414A1 (en) | 2002-11-22 | 2004-08-19 | Santora Ling C. | Method for reducing or preventing modification of a polypeptide in solution |
DE10256900A1 (en) | 2002-11-29 | 2004-06-24 | Nemod Immuntherapie Ag | Tumor-specific recognition molecules |
CN100450551C (en) | 2002-11-29 | 2009-01-14 | 中国医学科学院基础医学研究所 | Recombinated adeno-associated virus genes vaccine for prevention and cure of Alzheimer disease and use thereof |
EP1578947A4 (en) | 2002-12-02 | 2006-12-06 | Abgenix Inc | Antibodies directed to phospholipase a2 and uses thereof |
WO2004056318A2 (en) | 2002-12-19 | 2004-07-08 | New York University | Method for treating amyloid disease |
BR0317747A (en) | 2002-12-24 | 2005-11-22 | Neurochem Int Ltd | Method of concomitant therapeutic treatment of an individual, pharmaceutical composition, kit, use of a first agent and a second agent, and methods of preventing or treating amyloid-b-related disease, alzheimer's disease and mild cognitive impairment |
US20050031651A1 (en) | 2002-12-24 | 2005-02-10 | Francine Gervais | Therapeutic formulations for the treatment of beta-amyloid related diseases |
GB0230203D0 (en) | 2002-12-27 | 2003-02-05 | Domantis Ltd | Fc fusion |
AT413945B (en) | 2003-01-14 | 2006-07-15 | Mattner Frank Dr | Use of a compound having a binding capacity to an antibody specific for the natural N-terminal AB42 sequence, for preparing a vaccine for preventing and treating Alzheimer's disease |
US7662380B2 (en) | 2003-01-22 | 2010-02-16 | Takeda Pharmaceutical Company Limited | ZAQ ligand-1 antibodies and uses thereof |
EP2270051B1 (en) | 2003-01-23 | 2019-05-15 | Ono Pharmaceutical Co., Ltd. | Antibody specific for human PD-1 and CD3 |
WO2004065569A2 (en) | 2003-01-23 | 2004-08-05 | The Regents Of The University Of California | Multi-functional antibodies |
JP2006516639A (en) | 2003-02-01 | 2006-07-06 | ニユーララブ・リミテツド | Active immunization to generate antibodies against soluble A-beta |
WO2004068931A2 (en) | 2003-02-07 | 2004-08-19 | Protein Design Labs Inc. | Amphiregulin antibodies and their use to treat cancer and psoriasis |
US7575747B2 (en) | 2003-02-10 | 2009-08-18 | Applied Molecular Evolution | Aβ binding molecules |
US20040242845A1 (en) | 2003-02-21 | 2004-12-02 | Nicolau Yves Claude | Methods and compositions comprising supramolecular antigenic constructs and antibodies elicited against them |
US8663650B2 (en) | 2003-02-21 | 2014-03-04 | Ac Immune Sa | Methods and compositions comprising supramolecular constructs |
DE10307603A1 (en) | 2003-02-22 | 2004-09-02 | Rieter Ingolstadt Spinnereimaschinenbau Ag | textile machine |
US20060188951A1 (en) | 2003-02-24 | 2006-08-24 | In Hee Mook | Method for measuring the level of anti-beta-amyloid antibody in body fluids and diagnostic kit for alzheimer's disease using same |
US20040223970A1 (en) | 2003-02-28 | 2004-11-11 | Daniel Afar | Antibodies against SLC15A2 and uses thereof |
ES2335005T5 (en) | 2003-03-05 | 2013-04-17 | Halozyme, Inc. | Soluble hyaluronidase glycoprotein (shasegp), process to prepare it, uses and pharmaceutical compositions that comprise it |
US20060104968A1 (en) | 2003-03-05 | 2006-05-18 | Halozyme, Inc. | Soluble glycosaminoglycanases and methods of preparing and using soluble glycosaminogly ycanases |
US8545830B2 (en) | 2003-03-24 | 2013-10-01 | University Of Tennessee Research Foundation | Multi-functional polymeric materials and their uses |
EP1615946B1 (en) | 2003-03-26 | 2020-05-06 | Sudhir Paul | Proteolytic and covalent antibodies |
US20050129695A1 (en) | 2003-03-28 | 2005-06-16 | Marc Mercken | Anti-amyloid antibodies, compositions, methods and uses |
WO2004087733A2 (en) | 2003-03-28 | 2004-10-14 | New York University | Prevention and treatment of alzheimer amyloid deposition |
CN1189210C (en) | 2003-03-28 | 2005-02-16 | 万选才 | Coupling object between CB and biological active peptide or immunoglobhulin or immunological activity original as well as medication usage |
DK1625402T3 (en) | 2003-04-09 | 2014-03-10 | Canadian Blood Services | Detection, characterization and treatment of viral infection, and methods therefor |
EP1469312A1 (en) | 2003-04-18 | 2004-10-20 | Friedrich-Alexander-Universität Erlangen-Nürnberg | Diagnosis of Alzheimer's disease |
AU2003276107A1 (en) | 2003-04-24 | 2004-11-19 | Universitat Zurich | Method of monitoring immunotherapy |
EP1633316A4 (en) | 2003-05-06 | 2008-04-02 | Human Genome Sciences Inc | Antibodies that immunospecifically bind to trail receptors |
US20040223912A1 (en) | 2003-05-07 | 2004-11-11 | Montalto Michael Christopher | Compositions and methods for non-invasive imaging of soluble beta-amyloid |
ES2246178B1 (en) | 2003-05-08 | 2007-03-01 | Universidad De Zaragoza. | USE OF ANTIBODIES FOR THE TREATMENT OF AMYLOID DISEASES. |
EP1480041A1 (en) | 2003-05-22 | 2004-11-24 | Innogenetics N.V. | Method for the prediction, diagnosis and differential diagnosis of Alzheimer's disease |
WO2005018536A2 (en) | 2003-05-23 | 2005-03-03 | Human Genome Sciences, Inc. | Agonist antibodies that specifically bind the glucagon like peptide-1 receptor |
PE20050627A1 (en) | 2003-05-30 | 2005-08-10 | Wyeth Corp | HUMANIZED ANTIBODIES THAT RECOGNIZE THE BETA AMYLOID PEPTIDE |
CA2528182A1 (en) | 2003-06-06 | 2005-06-16 | Oncomax Acquisition Corp. | Antibodies specific for cancer associated antigen sm5-1 and uses thereof |
US7892751B2 (en) | 2003-06-09 | 2011-02-22 | Redox-Reactive Reagents Llc | Method of detecting or diagnosing of a neurodegenerative disease or condition |
JP4888876B2 (en) | 2003-06-13 | 2012-02-29 | 田平 武 | Recombinant adeno-associated virus vector for the treatment of Alzheimer's disease |
MXPA05013881A (en) | 2003-06-23 | 2008-02-13 | Genetics Inst Llc | Antibodies against interleukin-22 and uses therefor. |
EP2719394B1 (en) | 2003-06-30 | 2015-07-29 | Tel Aviv University Future Technology Development L.P. | Peptides for treating amyloid-associated diseases |
US20050009110A1 (en) | 2003-07-08 | 2005-01-13 | Xiao-Jia Chang | Methods of producing antibodies for diagnostics and therapeutics |
ATE444946T1 (en) | 2003-07-15 | 2009-10-15 | Ono Pharmaceutical Co | BRANCHED CARBOXYLIC ACID COMPOUND AND USE THEREOF |
WO2005012330A2 (en) | 2003-07-30 | 2005-02-10 | Brigham And Women's Hospital, Inc. | AMYLOID β-PEPTIDE AND METHODS OF USE |
US20050124016A1 (en) | 2003-08-01 | 2005-06-09 | Enh Research Institute | Antibodies specific for toxic amyloid beta protein oligomers |
WO2005014618A2 (en) | 2003-08-08 | 2005-02-17 | Immunomedics, Inc. | Bispecific antibodies for inducing apoptosis of tumor and diseased cells |
JP4239750B2 (en) | 2003-08-13 | 2009-03-18 | セイコーエプソン株式会社 | Microlens and microlens manufacturing method, optical device, optical transmission device, laser printer head, and laser printer |
WO2005018424A2 (en) | 2003-08-18 | 2005-03-03 | Research Foundation For Mental Hygiene, Inc. | Antibodies specific for fibrillar amyloid and a procedure to detect fibrillar amyloid deposits |
JP2007528723A (en) | 2003-08-22 | 2007-10-18 | メディミューン,インコーポレーテッド | Antibody humanization |
US20070031416A1 (en) | 2003-09-09 | 2007-02-08 | Takeda Pharmaceutical Company Limited | Use of antibody |
EP1660533A4 (en) | 2003-09-12 | 2009-10-21 | Univ California | Monoclonal antibodies specific for high molecular weight aggregation intermediates common to amyloids formed from proteins of differing sequence |
EP1516930A1 (en) | 2003-09-16 | 2005-03-23 | Georg-August Universität Göttingen | Cellular model of tauopathies for lead identification and drug discovery |
WO2005027965A1 (en) | 2003-09-24 | 2005-03-31 | Peter Krammer | Antibodies against annexins, use thereof for therapy and diagnosis. use of annexins for therapy and diagnosis. |
IL158287A0 (en) | 2003-10-07 | 2004-05-12 | Yeda Res & Dev | Antibodies to nik, their preparation and use |
ATE509034T1 (en) | 2003-10-07 | 2011-05-15 | Yeda Res & Dev | ANTIBODIES TO NIK, THEIR PRODUCTION AND USE |
WO2005037209A2 (en) | 2003-10-14 | 2005-04-28 | University Of South Florida | A method for the separation anti-amyloid beta antibody with amyloid beta peptide |
WO2005038457A1 (en) | 2003-10-15 | 2005-04-28 | Daiichi Pure Chemicals Co., Ltd. | Method of separating and assaying adiponectin multimer |
CA2543058A1 (en) | 2003-10-20 | 2005-05-12 | Envivo Pharmaceuticals, Inc. | Transgenic flies expressing mutant a.beta.42 |
JP2007535906A (en) | 2003-10-24 | 2007-12-13 | イステイチユート・デイ・リチエルケ・デイ・ビオロジア・モレコラーレ・ピ・アンジエレツテイ・エツセ・ピー・アー | Orthogonal gene switch |
EP2248830A1 (en) | 2003-11-04 | 2010-11-10 | Novartis Vaccines and Diagnostics, Inc. | Use of antagonist anti-CD40 antibodies for treatment of autoimmune and inflammatory diseases and organ transplant rejection |
US7807777B2 (en) | 2003-11-05 | 2010-10-05 | Immuno-Biological Laboratories Co., Ltd. | Marker peptide for alzheimer's disease |
WO2005047484A2 (en) | 2003-11-07 | 2005-05-26 | Ciphergen Biosystems, Inc. | Biomarkers for alzheimer's disease |
CA2545152A1 (en) | 2003-11-07 | 2005-05-26 | Howard J. Federoff | Compositions and methods of treating neurological diseases |
WO2005047860A2 (en) | 2003-11-08 | 2005-05-26 | Elan Pharmaceuticals, Inc. | Antibodies to alpha-synuclein |
KR100556660B1 (en) | 2003-11-11 | 2006-03-10 | 국립암센터 | Neutralizable epitope of hgf and neutralizing antibody binding to same |
WO2005052002A2 (en) | 2003-11-20 | 2005-06-09 | Massachusetts Institute Of Technology | Single-domain antibodies and uses thereof |
KR20060120161A (en) | 2003-11-28 | 2006-11-24 | 아스트라제네카 아베 | Antibodies |
JP4870348B2 (en) | 2003-12-04 | 2012-02-08 | 株式会社ペルセウスプロテオミクス | Antibody acquisition and antigen identification against cell surface antigens |
PL1691837T3 (en) | 2003-12-10 | 2012-11-30 | Squibb & Sons Llc | Ip-10 antibodies and their uses |
EA012984B1 (en) | 2003-12-17 | 2010-02-26 | Вайет | Immunogenic peptide carrier conjugates and method of producing same |
PT1711530E (en) | 2003-12-22 | 2009-10-22 | Glaxo Group Ltd | Nogo-a neutralising immunoglobulins for treatment of neurological diseases |
WO2005070965A2 (en) | 2004-01-21 | 2005-08-04 | Five Prime Therapeutics, Inc. | Pharmaceutical compositions containing antagonists to lrp4, lrp8 or megalin for treatment of diseases |
JP2007522119A (en) | 2004-01-28 | 2007-08-09 | キュリックス エーピーエス | Conjugates of amyloid protein as vaccines for amyloid-related diseases |
WO2005080986A1 (en) | 2004-02-18 | 2005-09-01 | University Of Iowa Research Foundation | Antibodies to phosphorylated tau, methods of making and methods of use |
WO2005080435A1 (en) | 2004-02-20 | 2005-09-01 | Immuno-Biological Laboratories Co., Ltd. | Monoclonal antibody and use thereof |
ES2375627T3 (en) | 2004-02-23 | 2012-03-02 | Eli Lilly And Company | ANTI-ABETA ANTIBODIES. |
WO2005105841A2 (en) | 2004-03-12 | 2005-11-10 | Human Genome Sciences, Inc. | Human g-protein chemokine receptor (ccr5) hdgnr10 |
EP1725870A1 (en) | 2004-03-18 | 2006-11-29 | Applied Research Systems ARS Holding N.V. | Anti-lipid rafts antibodies |
CN1314805C (en) | 2004-03-26 | 2007-05-09 | 中国人民解放军军事医学科学院放射与辐射医学研究所 | A novel APO and antibody thereof and preparation and use |
DK1730191T3 (en) | 2004-03-30 | 2011-10-17 | Glaxo Group Ltd | Immunoglobulin Binding to HOSM |
AU2005231364A1 (en) | 2004-04-02 | 2005-10-20 | Merck & Co., Inc. | Methods for detecting substances which bind to the amyloid precursor protein or beta amyloid fragments, and binding compounds |
US20060099211A1 (en) | 2004-04-12 | 2006-05-11 | Carmen Monthe | Safer, more potent human immunoglobulin preparations for treating Alzheimer's disease |
CN101022806A (en) | 2004-04-15 | 2007-08-22 | 萨马里坦药品公司 | Use of (4-alkylpiperazinyl)(phenyl) methanones in the treatment of alzheimer's disease |
AU2005233387B2 (en) | 2004-04-15 | 2011-05-26 | Glycofi, Inc. | Production of galactosylated glycoproteins in lower eukaryotes |
WO2005105998A1 (en) | 2004-04-27 | 2005-11-10 | Juridical Foundation The Chemo-Sero-Therapeutic Research Institute | HUMAN ANTIAMYLOID β PEPTIDE ANTIBODY AND ANTIBODY FRAGMENT THEREOF |
ITRM20040212A1 (en) | 2004-04-30 | 2004-07-30 | Lay Line Genomics Spa | NON-HUMAN TRANSGENIC ANIMAL AS A MODEL FOR NEURODEGENERATIVE DISEASES AND THEIR EARLY DIAGNOSIS. |
EP1755644A2 (en) | 2004-05-14 | 2007-02-28 | Northwestern University | Compositions comprising addl receptor syngap |
GR1005016B (en) | 2004-05-14 | 2005-10-11 | BIOMENTIKA@ΛΑΙΦ@ΣΑΙΕΝΣΙΣ@ΑΝΩΝΥΜΗ@ΕΤΑΙΡΕΙΑ@ΦΑΡΜΑΚΕΥΑΤΙΚΩΝ@ΠΡΟΙΟΝΤΩΝ@(συμμετέχει@σε@ποσοστό@50%)@Α | IgG & IgY ANTIBODIES DEVELOPED VERSUS THE HUMANIN SPECIALLY-SYNTHESIZED EPITOPE DERIVATIVE (BIO-ACTIVE 24-PEPTIDE) RELATED TO THE ALZHEIMER' S DISEASE AND DETECTION OF SAID BIO-ACTIVE 24- PEPTIDE BYSAME ANTIBODIES |
US20110142858A1 (en) | 2004-06-07 | 2011-06-16 | Ramot At Tel Aviv University Ltd. | Method of Passsive Immunization Against Disease or Disorder Charcterized by Amyloid Aggregation with Diminished Risk of Neuroinflammation |
US20060018896A1 (en) | 2004-06-10 | 2006-01-26 | University Of Leicester | Methods for treating conditions associated with lectin-dependent complement activation |
US20050276806A1 (en) | 2004-06-15 | 2005-12-15 | Advanced Biotherapy, Inc. | Treatment of autism |
CA2571272A1 (en) | 2004-06-18 | 2006-03-30 | He Xu | Anti-lympho- plus anti-monocytes globulin preparation for inhibiting immune responses |
SE0401601D0 (en) | 2004-06-21 | 2004-06-21 | Bioarctic Neuroscience Ab | Protofibril specific antibodies and uses thereof |
US8507206B2 (en) | 2004-07-02 | 2013-08-13 | Northwestern University | Monoclonal antibodies that target pathological assemblies of amyloid β (Abeta) |
WO2006005588A1 (en) | 2004-07-12 | 2006-01-19 | Geneprot, Inc. | Polypeptide species useful for the treatment of neurological disorders |
CN1721437A (en) | 2004-07-13 | 2006-01-18 | 中南大学 | Polypeptide antigen and antibody related to Tau protein |
AT500483B1 (en) | 2004-07-13 | 2006-01-15 | Mattner Frank Dr | Kit for prevention or treatment of Alzheimer's disease comprises means for inducing sequestration of amyloid beta in plasma and apheresis apparatus which exhibits an amyloid beta precursor protein receptor |
AT413946B (en) | 2004-07-13 | 2006-07-15 | Mattner Frank Dr | VACCINE AGAINST THE ALZHEIMER DISEASE |
WO2006006172A2 (en) | 2004-07-15 | 2006-01-19 | Ramot At Tel Aviv University Ltd. | Use of anti-amyloid agents for treating and typing pathogen infections |
WO2006014638A2 (en) | 2004-07-19 | 2006-02-09 | The General Hospital Corporation | ANTIBODIES TO CROSS-LINKED AMYLOID β OLIGOMERS |
ATE396990T1 (en) | 2004-07-28 | 2008-06-15 | Schering Corp | MACROCYCLIC INHIBITORS OF BETA SECRETASE |
US7807165B2 (en) | 2004-07-30 | 2010-10-05 | Rinat Neuroscience Corp. | Antibodies directed against amyloid-beta peptide and methods using same |
WO2006015621A1 (en) | 2004-08-09 | 2006-02-16 | Cellzome Ag | Treatment of neurodegenerative diseases by the use of scd4 inhibitors |
WO2006016644A1 (en) | 2004-08-11 | 2006-02-16 | Mitsubishi Chemical Corporation | Antibody and utilization of the same |
WO2006015976A1 (en) | 2004-08-11 | 2006-02-16 | Evotec Neurosciences Gmbh | Diagnostic and therapeutic use of a plasma membrane atpase |
DE102004039326A1 (en) | 2004-08-12 | 2006-02-16 | Abbott Gmbh & Co. Kg | Treatment of neurodegenerative diseases, specifically Alzheimer's disease and Down syndrome, uses a chemical/biological substance which inhibits fatty acid amide hydrolase without inhibiting cyclooxygenase-1 and/or 2 |
TWI374935B (en) | 2004-08-27 | 2012-10-21 | Pfizer Ireland Pharmaceuticals | Production of α-abeta |
WO2006039470A2 (en) | 2004-09-29 | 2006-04-13 | Centocor, Inc. | Anti- amyloid antibodies, compositions, methods and uses |
CA2582683A1 (en) | 2004-10-01 | 2006-04-13 | Max-Planck-Gesellschaft Zur Forderung Der Wissenschaften E.V. | Novel antibodies directed to the mammalian eag1 ion channel protein |
WO2006039327A2 (en) | 2004-10-01 | 2006-04-13 | Merck & Co., Inc. | Methods of treatment or prophylaxis of amyloidogenic diseases of the eye or optic nerve |
CN101027391A (en) | 2004-10-05 | 2007-08-29 | 惠氏公司 | Methods and compositions for improving recombinant protein production |
US20080063636A1 (en) | 2004-10-06 | 2008-03-13 | Hiroshi Mori | Variant Amyloid Protein |
EP1814917A2 (en) | 2004-10-13 | 2007-08-08 | Ablynx N.V. | Single domain camelide anti-amyloid beta antibodies and polypeptides comprising the same for the treatment and diagnosis of degenerative neural diseases such as alzheimer's disease |
WO2006047254A1 (en) | 2004-10-22 | 2006-05-04 | Regents Of The University Of Minnesota | Assemblies of oligomeric amyloid beta protein and uses thereof |
US7780963B2 (en) | 2004-10-25 | 2010-08-24 | Merck & Co., Inc. | Anti-ADDL antibodies and uses thereof |
WO2006047670A2 (en) | 2004-10-26 | 2006-05-04 | Wyeth | Methods for assessing antibodies to neurodegenerative disease-associated antigens |
CN101048662A (en) | 2004-10-28 | 2007-10-03 | 三光纯药株式会社 | Method of examining alzheimer s disease and diagnostic reagent |
WO2006050041A2 (en) | 2004-10-28 | 2006-05-11 | Ramot At Tel Aviv University Ltd. | Methods for reducing or inhibiting brain inflammation or for promoting neurogenesis |
US7709208B2 (en) | 2004-11-08 | 2010-05-04 | New York University | Methods for diagnosis of major depressive disorder |
AU2005304707A1 (en) | 2004-11-08 | 2006-05-18 | Nanobac Pharmaceuticals Incorporated | Methods and compositions for protein-hydroxy apatite complexes and their application in testing and modulating immunological system including a novel in vitro test for the detection of antibodies against calcium binding protein-hydroxy apatite complexes |
CN1772766A (en) | 2004-11-12 | 2006-05-17 | 中国科学院上海生命科学研究院 | Acetylcholinesterase monoclonal antibody related to resisting apoptosis and its use |
EP1824496A4 (en) | 2004-11-17 | 2008-07-16 | Joanne Mclaurin | Compositions comprising scyllo-inositol derivatives and methods to treat disorders of protein aggregation |
CA2589017A1 (en) | 2004-12-15 | 2006-06-22 | Neuralab Limited | Amyloid beta antibodies for use in improving cognition |
JP2008523815A (en) | 2004-12-15 | 2008-07-10 | エラン ファーマ インターナショナル リミテッド | Humanized amyloid beta antibody for use in improving cognition |
US20060240486A1 (en) | 2004-12-15 | 2006-10-26 | Johnson-Wood Kelly L | Immunoprecipitation-based assay for predicting in vivo efficacy of beta-amyloid antibodies |
PE20061152A1 (en) | 2004-12-15 | 2006-10-13 | Neuralab Ltd | HUMANIZED ANTIBODIES THAT RECOGNIZE THE BETA AMYLOID PEPTIDE |
WO2006066118A2 (en) | 2004-12-15 | 2006-06-22 | Neuralab Limited | Contextual fear test for predicting efficacy of alzheimer immunotherapeutic treatment |
JP2006166879A (en) | 2004-12-20 | 2006-06-29 | Japan Health Science Foundation | Ab-dip, and preventing and treating agent of alzheimer's disease |
GB0428084D0 (en) * | 2004-12-22 | 2005-01-26 | Nokia Corp | Method for producing authentication information |
WO2006067792A2 (en) | 2004-12-22 | 2006-06-29 | Yeda Research And Development Co. Ltd. At The Weizmann Institute Of Science | Aldolase autoantigens useful in diagnosis and treatment of alzheimer's disease |
WO2006069081A2 (en) | 2004-12-22 | 2006-06-29 | Washington University In St. Louis | USE OF ANTI-Aβ ANTIBODY TO TREAT TRAUMATIC BRAIN INJURY |
MY146381A (en) | 2004-12-22 | 2012-08-15 | Amgen Inc | Compositions and methods relating relating to anti-igf-1 receptor antibodies |
EP1676859A1 (en) | 2004-12-30 | 2006-07-05 | Pevion Biotech Ltd. | Immunogenic compositions of cyclic peptides derived from the beta-amyloid peptide |
EP1853299A4 (en) | 2005-01-14 | 2009-11-11 | Univ California | Compositions and methods for inhibiting drusen formation and for diagnosing or treating drusen-related disorders |
CA2589860A1 (en) | 2005-01-24 | 2006-08-03 | Amgen Inc. | Humanized anti-amyloid antibody |
GT200600031A (en) | 2005-01-28 | 2006-08-29 | ANTI-BETA ANTIBODY FORMULATION | |
JP2006213621A (en) | 2005-02-02 | 2006-08-17 | Japan Health Science Foundation | Adoplin protein and utilization of the same |
US7790363B2 (en) | 2005-02-07 | 2010-09-07 | Abbott Laboratories Inc. | Diagnostic test for vitamin B12 |
US20070082350A1 (en) | 2005-02-09 | 2007-04-12 | Philip Landfield | Assay and method for diagnosing and treating alzheimer's disease |
US7731962B2 (en) | 2005-02-14 | 2010-06-08 | Merck & Co., Inc. | Anti-ADDL monoclonal antibody and use thereof |
US7700099B2 (en) | 2005-02-14 | 2010-04-20 | Merck & Co., Inc. | Non-immunostimulatory antibody and compositions containing the same |
GB0503434D0 (en) | 2005-02-18 | 2005-03-30 | Senexis Ltd | Amyloid-binding peptides, analogues and uses thereof |
CN103936859A (en) | 2005-03-03 | 2014-07-23 | 免疫医疗公司 | Humanized L243 antibodies |
EP1869084A2 (en) | 2005-03-04 | 2007-12-26 | Biogen Idec MA Inc. | Methods of humanizing immunoglobulin variable regions through rational modification of complementarity determining residues |
ES2340414T3 (en) * | 2005-03-05 | 2010-06-02 | ABBOTT GMBH & CO. KG | METHOD OF SELECTION, PROCESS FOR PURIFICATION OF NON-DIFFUSIBLE A-BETA OLIGOMERS, SELECTIVE ANTIBODIES AGAINST SUCH NON-DIFFUSABLE A-BETA OLIGOMERS AND A PROCESS FOR THE MANUFACTURE OF SUCH ANTIBODIES. |
WO2006096529A2 (en) | 2005-03-07 | 2006-09-14 | Novartis Ag | Genes involved in neurodegenerative conditions |
ES2259270B1 (en) | 2005-03-09 | 2007-11-01 | Consejo Superior De Investigaciones Cientificas | IN VITRO DIAGNOSTIC METHOD OF ALZHEIMER'S DISEASE BY A MONOCLONAL ANTIBODY. |
WO2006099543A2 (en) | 2005-03-15 | 2006-09-21 | The Regents Of The University Of California | Methods for assessing antibody-mediated cytotoxicity |
WO2006100679A2 (en) | 2005-03-22 | 2006-09-28 | Quark Pharmaceuticals, Inc. | Recombinant antibodies against human type ii transglutaminase and uses thereof |
JP2006265189A (en) | 2005-03-24 | 2006-10-05 | Kyoto Univ | beta-AMYLOID PEPTIDE AND METHOD FOR SCREENING THERAPEUTIC AGENT OR PROPHYLACTIC AGENT FOR ALZHEIMER'S DISEASE USING THE SAME |
ES2318918B1 (en) | 2005-04-01 | 2010-02-16 | Biotherapix Molecular Medicines, S.L.U. | HUMAN ANTIBODIES WITH CAPACITY OF UNION TO THE BETA-AMYLOID PEPTIDE AND ITS APPLICATIONS. |
US8227194B2 (en) | 2005-04-08 | 2012-07-24 | University Of Maryland, Baltimore | Monoclonal antibodies with binding specificity for response gene to complement 32 (RGC-32) |
US20060241038A1 (en) | 2005-04-20 | 2006-10-26 | Eisai Co., Ltd. | Therapeutic agent for Abeta related disorders |
RU2007143302A (en) | 2005-04-22 | 2009-05-27 | Дженентек, Инк. (Us) | METHOD FOR TREATING DEMENTIA OR ALZHEIMER'S DISEASE ANTIBODIES TO CD20 |
UY29504A1 (en) | 2005-04-29 | 2006-10-31 | Rinat Neuroscience Corp | DIRECTED ANTIBODIES AGAINST BETA AMYLOID PEPTIDE AND METHODS USING THE SAME. |
WO2006119449A2 (en) | 2005-05-04 | 2006-11-09 | Vectorlogics, Inc. | Modified adenovirus containing a stabilized antibody |
ATE535252T1 (en) | 2005-05-05 | 2011-12-15 | Merck Sharp & Dohme | PEPTIDE CONJUGATE COMPOSITIONS AND METHODS FOR PREVENTING AND TREATING ALZHEIMER'S DISEASE |
WO2006125830A2 (en) | 2005-05-27 | 2006-11-30 | Evotec Neurosciences Gmbh | Kcnn3 as diagnostic and therapeutic target for neurodegenerative diseases |
EP1907566A4 (en) | 2005-05-27 | 2008-12-10 | Memory Pharm Corp | Transgenic alzheimer's mouse model vectors and uses thereof |
EP2298813A3 (en) | 2005-06-06 | 2012-03-21 | Wyeth LLC | Anti-TRKB monoclonal antibodies and uses thereof |
EA201100177A1 (en) | 2005-06-17 | 2011-06-30 | Элан Фарма Интернэшнл Лимитед | METHODS OF CLEANING ANTIBODIES TO β-AMYLOID |
WO2006137354A1 (en) | 2005-06-21 | 2006-12-28 | Medical & Biological Laboratories Co., Ltd. | Antibody having inhibitory effect on amyloid fibril formation |
TW200726482A (en) | 2005-06-30 | 2007-07-16 | Merck & Co Inc | Method for preparing a covalently cross linked oligomer of amyloid beta peptides |
TW200726774A (en) | 2005-06-30 | 2007-07-16 | Merck & Co Inc | Composition and method for producing stable amyloid beta oligomers |
EP2394661A1 (en) | 2005-07-08 | 2011-12-14 | Biogen Idec MA Inc. | Sp35 antibodies and uses thereof |
US20090297534A1 (en) | 2005-07-13 | 2009-12-03 | Sudhir Paul | Catalytic Immunoglobulins BBK32 and Uses Therefor |
WO2007011834A2 (en) | 2005-07-15 | 2007-01-25 | The Regents Of The University Of California | Compounds and method for the diagnosis and treatment of amyloid associated diseases |
AU2006279896A1 (en) | 2005-08-10 | 2007-02-22 | Oklahoma Medical Research Foundation | Truncated memapsin 2 for use for treating Alzheimer's disease |
CN105017423B (en) | 2005-08-11 | 2019-06-04 | 阿皮·马托西安-罗杰斯 | TCR-V- beta related peptides for treatment of autoimmune diseases and diagnosis |
KR20080068004A (en) | 2005-08-15 | 2008-07-22 | 아라나 테라퓨틱스 리미티드 | Engineered antibodies with new world primate framework regions |
WO2007022416A2 (en) | 2005-08-18 | 2007-02-22 | Ramot At Tel Aviv University Ltd. | Single chain antibodies against beta-amyloid peptide |
US7612181B2 (en) | 2005-08-19 | 2009-11-03 | Abbott Laboratories | Dual variable domain immunoglobulin and uses thereof |
JP2007077103A (en) | 2005-09-16 | 2007-03-29 | Yokohama City Univ | Prophylactic or therapeutic agent for alzheimer's disease |
WO2007040437A1 (en) | 2005-10-03 | 2007-04-12 | Astrazeneca Ab | Fusion proteins having a modulated half-life in plasma |
EP1940881B1 (en) | 2005-10-11 | 2016-11-30 | Amgen Research (Munich) GmbH | Compositions comprising cross-species-specific antibodies and uses thereof |
US20070092508A1 (en) | 2005-10-21 | 2007-04-26 | Recombiant Technologies, Llc | Detoxification depot for Alzheimer's disease |
WO2007047995A2 (en) | 2005-10-21 | 2007-04-26 | Catalyst Biosciences, Inc. | Modified proteases that inhibit complement activation |
CA2626783A1 (en) | 2005-10-21 | 2007-05-03 | Merck & Co., Inc. | Anti-addl monoclonal antibody and use thereof |
EP1957538B1 (en) | 2005-11-01 | 2013-05-22 | Novartis AG | Uses of anti-cd40 antibodies |
US20090181008A1 (en) | 2005-11-10 | 2009-07-16 | Satoris, Inc. | Methods of treating alzheimer's disease |
US8316104B2 (en) | 2005-11-15 | 2012-11-20 | California Institute Of Technology | Method and apparatus for collaborative system |
US20080014596A1 (en) | 2005-11-16 | 2008-01-17 | Jasna Jerecic | ADDL Binding to Hippocampal Neurons |
AU2006318537A1 (en) | 2005-11-22 | 2007-05-31 | The Trustees Of The University Of Pennsylvania | Antibody treatment of Alzheimer's and related diseases |
CA2631195C (en) | 2005-11-30 | 2016-04-05 | Abbott Laboratories | Monoclonal antibodies against amyloid beta protein and uses thereof |
CN101432302A (en) * | 2005-11-30 | 2009-05-13 | 艾博特公司 | Anti-a globulomer antibodies, antigen-binding moieties thereof, corresponding hybridomas, nucleic acids, vectors, host cells, methods of producing said antibodies, compositions comprising said antibod |
MX2008006957A (en) | 2005-11-30 | 2008-10-20 | Abbott Lab | Methods of preparation of recombinant forms of human beta-amyloid protein and uses of these proteins. |
WO2007064919A2 (en) | 2005-12-02 | 2007-06-07 | Genentech, Inc. | Binding polypeptides with restricted diversity sequences |
WO2007067512A2 (en) | 2005-12-08 | 2007-06-14 | Merck & Co., Inc. | Method for identifying modulators of adprh useful for treating alzheimer's disease |
PT1960428E (en) | 2005-12-12 | 2011-10-07 | Hoffmann La Roche | Antibodies against amyloid beta with glycosylation in the variable region |
DK1959991T3 (en) | 2005-12-12 | 2013-06-17 | Ac Immune Sa | THERAPEUTIC VACCINE |
PT2361638E (en) | 2005-12-12 | 2014-04-17 | Ac Immune Sa | A beta 1-42 specific monoclonal antibodies with therapeutic properties |
EP1973948B1 (en) | 2005-12-15 | 2015-02-11 | Genentech, Inc. | Methods and compositions for targeting polyubiquitin |
US20080058276A1 (en) | 2006-01-13 | 2008-03-06 | Cornell Research Foundation, Inc. | Alzheimer's disease therapeutics based on pin-1 catalyzed conformational changes in phosphorylated amyloid precursor protein |
EP1811304A1 (en) | 2006-01-18 | 2007-07-25 | Friedrich-Alexander-Universität Erlangen-Nürnberg | Large Aß-peptide binding particles (LAPS) in diagnosis and therapy of Alzheimer's dementia |
CN1329413C (en) | 2006-01-23 | 2007-08-01 | 南京医科大学 | Antibody for treating or preventing senile dementia, its expression vector and application in drug preparation |
PT1981540E (en) | 2006-01-30 | 2013-05-07 | Grifols Therapeutics Inc | Method of treatment and prophylaxis of diseases related to amyloid deposition using igm |
GB0601976D0 (en) | 2006-02-01 | 2006-03-15 | Merck Sharp & Dohme | Proteins |
WO2007088712A1 (en) | 2006-02-02 | 2007-08-09 | National University Corporation Nagoya University | Neuronal cell death inhibitor and screening method |
US20080045499A1 (en) | 2006-02-06 | 2008-02-21 | Elan Pharmaceuticals, Inc. | Preferential Inhibition of Presenilin-1 |
WO2007090872A2 (en) | 2006-02-09 | 2007-08-16 | Novartis Ag | Antibodies against secreted fri zzled related protein-4 (sfrp-4 ) |
EP1991252A2 (en) | 2006-02-21 | 2008-11-19 | Oklahoma Medical Research Foundation | Treatment of alzheimer's disease with inhibitors of apoe binding to apoe receptor |
CN101421299A (en) | 2006-02-22 | 2009-04-29 | 株式会社林原生物化学研究所 | Peptide vaccine for inducing production of anti-amyloid- -peptide antibody |
US20070196367A1 (en) | 2006-02-22 | 2007-08-23 | Valentin Dinu | Methods of preventing and treating Alzheimer's disease, age related macular degeneration and other diseases involving extra-cellular debris through the inhibition of the complement system |
MX2008010633A (en) | 2006-02-24 | 2008-09-25 | Chiesi Farma Spa | Anti-amyloid immunogenic compositions, methods and uses. |
US20100196932A1 (en) | 2006-03-02 | 2010-08-05 | The Board Of Trustees Of The University Of Illinois | Yeast Reporter System |
EP2514823B1 (en) | 2006-03-03 | 2018-05-02 | ProMIS Neurosciences Inc. | Methods and compositions to treat and detect misfolded-SOD1 mediated diseases |
WO2007109107A2 (en) | 2006-03-17 | 2007-09-27 | The Trustees Of Columbia University In The City Of New York | Atf4 as a therapeutic target in alzheimers disease and other neurological disorders |
WO2007109749A2 (en) | 2006-03-21 | 2007-09-27 | Wyeth | Methods for preventing and treating amyloidogenic diseases |
EP2325209A3 (en) | 2006-03-23 | 2011-08-03 | BioArtic Neuroscience AB | Improved protofibril selective antibodies and the use thereof |
EP2007385A4 (en) | 2006-03-23 | 2010-08-18 | Sinai School Medicine | Cardiovascular composition and use the same for the treatment of alzheimers disease |
CA2541522A1 (en) | 2006-03-28 | 2007-09-28 | E. Rick Preddie | A double elisa for alzheimer's disease (ad) with applications for therapeutic vaccines to stop the disease |
FR2899107B1 (en) | 2006-03-30 | 2008-06-13 | Neurokin Entpr Unipersonnelle | USE OF (S) -ROSCOVITINE FOR THE MANUFACTURE OF A MEDICINAL PRODUCT |
CN103539857A (en) | 2006-03-30 | 2014-01-29 | 葛兰素集团有限公司 | Antibodies against amyloid-beta peptide |
US7851228B2 (en) | 2006-03-31 | 2010-12-14 | The Regents Of The University Of California | Methods for screening agents that modulate presenilin activity and A-β production |
JP4790013B2 (en) | 2006-04-13 | 2011-10-12 | エーディア株式会社 | Method and diagnostic reagent used for assay of Alzheimer's disease by measuring blood degradation rate of β-amyloid |
CN101058608B (en) | 2006-04-21 | 2011-02-23 | 杜如昱 | Human anti-Abeta(1-32) amyloid antibody, purifying method and use thereof |
KR101132293B1 (en) | 2006-04-21 | 2012-04-05 | 주식회사 피플바이오 | Method for Differentially Detecting a Multimeric Form from a Monomeric Form of Multimer-Forming Polypeptides Through Three-Dimensional Interactions |
WO2007129457A1 (en) | 2006-04-25 | 2007-11-15 | The University Of Tokyo | Therapeutic agents for alzheimer's disease and cancer |
GB0608386D0 (en) | 2006-04-27 | 2006-06-07 | Senexis Ltd | Compounds |
US20070259831A1 (en) | 2006-04-27 | 2007-11-08 | Carlezon William A Jr | Treatment and screening methods for promoting neurogenesis |
CA2650704A1 (en) | 2006-04-28 | 2007-11-08 | Northwestern University | Salts of pyridazine compounds |
JP2007300856A (en) | 2006-05-11 | 2007-11-22 | Hiroshi Mori | Amyloid protein imitation |
US20070292895A1 (en) | 2006-05-19 | 2007-12-20 | Xiao-Ping Shi | Assays and methods to detect beta-secretase and its activity in body fluids and tissue extracts |
WO2008042024A2 (en) | 2006-06-01 | 2008-04-10 | Elan Pharmaceuticals, Inc. | Neuroactive fragments of app |
US7427342B2 (en) | 2006-06-02 | 2008-09-23 | General Electric Company | Method and apparatus for shifting current distribution in electrodeionization systems |
US7479550B2 (en) | 2006-06-02 | 2009-01-20 | The Board Of Regents Of The University Of Texas System | Amyloid β gene vaccines |
JP4933159B2 (en) | 2006-06-02 | 2012-05-16 | 国立大学法人金沢大学 | Diagnostic method for Alzheimer's disease |
US20090264359A1 (en) | 2006-06-16 | 2009-10-22 | Cornelis Petrus Maria Van Kessel | Fplr-1 inhibitors for use in diseases involving amyloid-induced inflammatory events (flipr and flipr-like) and immunecomplex-mediated diseases |
TW200815469A (en) | 2006-06-23 | 2008-04-01 | Astrazeneca Ab | Compounds |
WO2008002893A2 (en) | 2006-06-29 | 2008-01-03 | Centocor, Inc. | Anti-amyloid antibodies, compositions, methods and uses |
KR20090026275A (en) | 2006-07-06 | 2009-03-12 | 로스캄프 리서치 엘엘씨 | Compounds and combinations thereof for inhibiting beta-amyloid production and methods of use thereof |
US20080012103A1 (en) | 2006-07-13 | 2008-01-17 | Foster Robert H | Emi absorbing gap filling material |
WO2008008463A2 (en) | 2006-07-14 | 2008-01-17 | Trustees Of Columbia University In The City Of New York | METHODS AND COMPOSITIONS FOR DETECTING AND QUANTIFYING SAPPβ |
EP2046833B9 (en) | 2006-07-14 | 2014-02-19 | AC Immune S.A. | Humanized antibody against amyloid beta |
ATE426174T1 (en) | 2006-07-28 | 2009-04-15 | Vista Ventures Gmbh | METHOD FOR DETECTING AMYLOID-BETA OLIGOMERS IN BODY FLUIDS |
US7705475B2 (en) | 2006-08-03 | 2010-04-27 | Stats Chippac Ltd. | Integrated circuit package system |
CA2657847A1 (en) | 2006-08-04 | 2008-02-07 | Lonza Biologics Plc | Method for predicting protein aggregation and designing aggregation inhibitors |
WO2008021296A2 (en) | 2006-08-14 | 2008-02-21 | Thymon, L.L.C. | Compositions and methods for the treatment and prophylaxis of alzheimer's disease |
US7741446B2 (en) | 2006-08-18 | 2010-06-22 | Armagen Technologies, Inc. | Fusion antibodies that cross the blood-brain barrier in both directions |
WO2008051326A2 (en) | 2006-08-21 | 2008-05-02 | President And Fellows Of Harvard College | Identification of contactins and l1- cams as ligands for the amyloid precursor protein |
WO2008070229A2 (en) | 2006-08-28 | 2008-06-12 | Case Western Reserve University | Detection of pathogenic aggregates of protein in a sample by homologous elisa |
JP2010502623A (en) | 2006-08-31 | 2010-01-28 | バイオジェン・アイデック・エムエイ・インコーポレイテッド | Methods for peripheral administration of Nogo receptor polypeptides |
US8372399B2 (en) | 2006-08-31 | 2013-02-12 | Cardiac Pacemakers, Inc. | Bispecific antibodies and agents to enhance stem cell homing |
US20080118938A1 (en) | 2006-09-06 | 2008-05-22 | Lisbell Estrada | Methods and Compositions for the Detection of Protein Folding Disorders |
CA2662723A1 (en) | 2006-09-08 | 2008-03-13 | Vib Vzw | Means and methods for the production of amyloid oligomers |
WO2008030251A1 (en) | 2006-09-08 | 2008-03-13 | Georgetown University | Deglycosylated anti-amyloid beta antibodies |
ES2307396B1 (en) | 2006-09-14 | 2009-09-30 | Fundacion Para Investigaciones Neurologicas (Fin) | SYSTEMS FOR THE ELIMINATION OF NEUROTOXIC SUBSTANCES CAUSING NEURODEGENERATIVE DISEASES THROUGH THEIR SELECTIVE ATTACKING BY IMMUNOFINITY IN THE CIRCULATING CEPHALO-RAQUIDEO LIQUID. |
US7375190B2 (en) | 2006-09-19 | 2008-05-20 | National Yang-Ming University | Recombinant protein and method of screening for agents that modulate polypeptide aggregation |
BRPI0719763A2 (en) | 2006-10-02 | 2014-01-28 | Ac Immune Sa | ANTIBODY, NUCLEIC ACID MOLECULE, EXPRESSION VECTOR, CELL, COMPOSITION, MIXTURE, USE OF A CHEMICAL ANTIBODY OR A FRAGMENT OF THE SAME OR A FRAGMENT OF THE SAME AND / OR A PART OR A FUNCTION OR A PART OR A FUNCTION MIXTURE, METHODS FOR PREPARING A PHARMACEUTICAL COMPOSITION OR MIXTURE, TO PREVENT, TREAT OR RELIEVE THE EFFECTS OF A DISEASE, DIAGNOSTIC DISEASE, OR CONDITION ASSOCIATED WITH A DETERMINATION OF THE DETERMINATION OF CAROGUE DETERMINATION IN A FABRIC AND / OR BODY FLUIDS, TEST KITS FOR DISEASE AND DIAGNOSTIC TESTING AND CONDITIONS ASSOCIATED WITH AMYLOID, LIGHT CHAIN VARIABLE REGION, HEAVY CIRCLE, GENELE DESENGER, GENELE DIAGET LINES PRE-FORMED BETA-AMYLOID FIBERS TO PREVENT ABETA-INDUCED NEURON DEGRADATION, TO DIAGNOSE A PREDISPOSITION TO A DISEASE OR CO NDITION ASSOCIATED WITH AMYLOID IN A PATIENT, TO MONITOR MINIMUM RESIDUAL DISEASE IN A PATIENT, TO PROGNOSE LIABILITY OF A PATIENT WHO IS TREATED WITH AN ANTIBODY OR A COMPOSITION OF VACCINE, TO REDUCE THE CARBON LOAD TO THE CIRCLE THE NUMBER OF PLATES IN AN ANIMAL BRAIN, TO REDUCE THE TOTAL SOLUBLE OPEN QUANTITY IN AN ANIMAL BRAIN AND TO HOLD OR INCREASE COGNITIVE MEMORY CAPACITY IN A MAMMALIAN. |
WO2008045962A2 (en) | 2006-10-10 | 2008-04-17 | Mayo Foundation For Medical Education And Research | Methods and materials related to anti-a (beta) antibodies |
US9382327B2 (en) | 2006-10-10 | 2016-07-05 | Vaccinex, Inc. | Anti-CD20 antibodies and methods of use |
JP5153114B2 (en) | 2006-10-12 | 2013-02-27 | 知宏 千葉 | Novel method for detecting Alzheimer's disease |
US20080113444A1 (en) | 2006-10-17 | 2008-05-15 | Pray Todd R | Method for detecting oligermization of soluble amyloid beta oligomers |
GB0620735D0 (en) | 2006-10-18 | 2006-11-29 | Ares Trading Sa | Proteins |
KR100883132B1 (en) | 2006-10-24 | 2009-02-10 | 재단법인서울대학교산학협력재단 | A cleavage agent selectively acting on soluble assembly of amyloidogenic peptide or protein |
NZ613356A (en) | 2006-10-27 | 2015-02-27 | Abbvie Biotechnology Ltd | Crystalline anti-htnfalpha antibodies |
WO2008064244A2 (en) | 2006-11-20 | 2008-05-29 | The Trustees Of Columbia University In The City Of New York | Phosphoinositide modulation for the treatment of neurodegenerative diseases |
US20100056487A1 (en) | 2006-11-20 | 2010-03-04 | Satori Pharmaceuticals, Inc. | Modulators of amyloid-beta production |
KR101450356B1 (en) | 2006-11-24 | 2014-10-15 | 에이씨 이뮨 에스.에이. | N-(methyl)-1h-pyrazol-3-amine, n-(methyl)-pyridin-2-amine and n-(methyl)-thiazol-2-amine derivatives for the treatment of diseases associated with amyloid or amyloid-like proteins, like e.g. alzheimer's |
US8455626B2 (en) | 2006-11-30 | 2013-06-04 | Abbott Laboratories | Aβ conformer selective anti-aβ globulomer monoclonal antibodies |
WO2008143708A2 (en) | 2006-12-07 | 2008-11-27 | Mayo Foundation For Medical Education And Research | Methods and materials related to anti-amyloid antibodies |
PE20081477A1 (en) | 2006-12-11 | 2008-10-18 | Hoffmann La Roche | LYOPHILIZED FORMULATION MAB ABETA |
US20100028333A1 (en) | 2006-12-15 | 2010-02-04 | Getty Krista L | Receptor for amyloid beta and uses thereof |
DK2104682T3 (en) | 2007-01-11 | 2017-01-16 | Michael Bacher | DIAGNOSIS AND TREATMENT OF ALZHEIMER'S AND OTHER DEMENTIA DISEASES |
WO2008140639A2 (en) | 2007-02-08 | 2008-11-20 | Oligomerix, Inc. | Biomarkers and assays for alzheimer's disease |
KR100806914B1 (en) | 2007-02-14 | 2008-02-22 | 경북대학교 산학협력단 | 2 Novel use of lipocalin 2 for preventing and treating neurodegenerative disease |
EP2124952A2 (en) | 2007-02-27 | 2009-12-02 | Abbott GmbH & Co. KG | Method for the treatment of amyloidoses |
WO2008104385A1 (en) | 2007-02-27 | 2008-09-04 | Abbott Gmbh & Co. Kg | Method for the treatment of amyloidoses |
GB0704394D0 (en) | 2007-03-07 | 2007-04-11 | Senexis Ltd | Compounds |
US20090022728A1 (en) | 2007-03-09 | 2009-01-22 | Rinat Neuroscience Corporation | Methods of treating ophthalmic diseases |
EP1978035A1 (en) | 2007-04-05 | 2008-10-08 | Hans-Knöll-Institut Leibniz-Institut für Naturstoff-Forschung | Anti-amyloid antibodies and their use in diagnosis and therapy of amyloid diseases |
JP2010523602A (en) | 2007-04-12 | 2010-07-15 | ワラタ ファーマシューティカルズ, インコーポレイテッド | Use of cyclohexanehexol derivatives in the treatment of eye diseases |
JP2011526240A (en) | 2007-04-18 | 2011-10-06 | ヤンセン アルツハイマー イミュノセラピー | Prevention and treatment of cerebral amyloid angiopathy |
CA2683053A1 (en) | 2007-04-19 | 2008-10-30 | Vib Vzw | Oligonucleotide compositions for the treatment of alzheimer's disease |
US9217036B2 (en) | 2007-04-26 | 2015-12-22 | Yale University | Prion protein as a receptor for amyloid-β oligomers |
US20090232801A1 (en) | 2007-05-30 | 2009-09-17 | Abbot Laboratories | Humanized Antibodies Which Bind To AB (1-42) Globulomer And Uses Thereof |
US20090175847A1 (en) | 2007-05-30 | 2009-07-09 | Abbott Laboratories | Humanized antibodies to ab (20-42) globulomer and uses thereof |
US20100178658A1 (en) | 2007-05-30 | 2010-07-15 | The Regents Of The University Of Michigan | Screening assays for inhibitors of beta amyloid peptide ion channel formation |
KR20160017126A (en) | 2007-06-12 | 2016-02-15 | 에이씨 이뮨 에스.에이. | Monoclonal anti beta amyloid antibody |
PE20090766A1 (en) | 2007-06-12 | 2009-07-09 | Ac Immune Sa | HUMANIZED IGG1 ANTIBODY |
EP2009445A1 (en) | 2007-06-29 | 2008-12-31 | Institut Pasteur | Use of a camelid single-domain antibody for detecting an oligomeric form of an amyloid beta peptide and its applications |
CN101084909A (en) | 2007-07-03 | 2007-12-12 | 福建医科大学附属协和医院 | New use of ginsenoside Rg1 |
WO2009009768A2 (en) | 2007-07-12 | 2009-01-15 | Acumen Pharmaceuticals, Inc. | METHODS OF INHIBITING THE FORMATION OF AMYLOID-β DIFFUSABLE LIGANDS USING A ACYLHYDRAZIDE COMPOUDS |
US8962677B2 (en) | 2007-07-12 | 2015-02-24 | Acumen Pharmaceuticals, Inc. | Methods of restoring cognitive ability using non-peptidic compounds |
CA2692775C (en) | 2007-07-12 | 2015-03-24 | Acumen Pharmaceuticals, Inc. | Methods of enhancing cognitive function using non-peptidic compounds |
CA2692773A1 (en) | 2007-07-12 | 2009-01-15 | Acumen Pharmaceuticals, Inc. | Methods of modifying amyloid .beta. oligomers using non-peptidic compounds |
SG178809A1 (en) | 2007-10-05 | 2012-03-29 | Genentech Inc | Use of anti-amyloid beta antibody in ocular diseases |
CN105169388A (en) | 2007-10-05 | 2015-12-23 | 基因技术公司 | Humanized antibody |
GB0719559D0 (en) | 2007-10-05 | 2007-11-14 | Senexis Ltd | Compounds |
CA2701793C (en) | 2007-10-05 | 2017-04-25 | Genentech, Inc. | Use of anti-amyloid beta antibody in ocular diseases |
CN101152576A (en) | 2007-10-15 | 2008-04-02 | 王延江 | Medicament for preventing and controlling alzheimer's disease |
CR20170001A (en) | 2008-04-28 | 2017-08-10 | Genentech Inc | ANTI FACTOR D HUMANIZED ANTIBODIES |
US20100173828A1 (en) | 2008-07-25 | 2010-07-08 | Abbott Gmbh & Co. Kg | Aß(X - 38 .. 43) oligomers, and processes, compositions, and uses thereof |
CN102203124A (en) | 2008-07-25 | 2011-09-28 | 雅培制药有限公司 | Amyloid ss peptide analogues, oligomers thereof, processes for preparing and compositions comprising said analogues or oligomers, and their uses |
EP2401300B1 (en) * | 2009-02-25 | 2018-01-10 | Academia Sinica | ANTI-C[epsilon]mX ANTIBODIES CAPABLE OF BINDING TO HUMAN mIgE ON B LYMPHOCYTES |
US8987419B2 (en) | 2010-04-15 | 2015-03-24 | AbbVie Deutschland GmbH & Co. KG | Amyloid-beta binding proteins |
MX358739B (en) | 2010-08-14 | 2018-09-03 | Abbvie Inc Star | Amyloid-beta binding proteins. |
-
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Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2004067561A1 (en) | 2003-01-31 | 2004-08-12 | Abbott Gmbh & Co. Kg | Amyloid-$g(b)(1-42) oligomers, derivatives thereof, antibodies for the same, method for production and use therof |
Non-Patent Citations (4)
Title |
---|
BARGHORN ET AL., J NEUROCHEM, vol. 95, 2005, pages 834 - 847 |
SELKOE, NEURON, vol. 32, 2001, pages 177 - 180 |
SERGEANT ET AL., J NEUROCHEM, vol. 85, 2003, pages 1581 - 1591 |
THAL ET AL., J NEUROPATHOL. EXP NEUROL, vol. 58, 1999, pages 210 - 216 |
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