CN102016583B - Glycan based array and uses thereof - Google Patents

Glycan based array and uses thereof Download PDF

Info

Publication number
CN102016583B
CN102016583B CN200980112434.8A CN200980112434A CN102016583B CN 102016583 B CN102016583 B CN 102016583B CN 200980112434 A CN200980112434 A CN 200980112434A CN 102016583 B CN102016583 B CN 102016583B
Authority
CN
China
Prior art keywords
disease
candidate
antibody
tissue
target
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN200980112434.8A
Other languages
Chinese (zh)
Other versions
CN102016583A (en
Inventor
王慧茹
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Jiangsu Mingjian Biopharmaceutical Co ltd
Original Assignee
B & H Biotechnologies
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by B & H Biotechnologies filed Critical B & H Biotechnologies
Publication of CN102016583A publication Critical patent/CN102016583A/en
Application granted granted Critical
Publication of CN102016583B publication Critical patent/CN102016583B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/5308—Immunoassay; Biospecific binding assay; Materials therefor for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2400/00—Assays, e.g. immunoassays or enzyme assays, involving carbohydrates

Abstract

The present disclosure discloses simple and efficient glycan- or carbohydrate-based processes or methods for the rapid identification of biological markers and therapeutic targets especially glycan-related targets of infectious diseases, cancers, autoimmune diseases, allergies, inflammation, toxicity, obesity and/or other disorders of humans, animals, plants and other organisms. Therefore, novel methods and products for the diagnosis, prevention, and treatment of such diseases obtainable based on these therapeutic targets can be developed.

Description

Chip and its application based on polysaccharide
Priority
Entitled " the molecular mimicry chip and its application based on polysaccharide " that application claims on April 8th, 2008 submits to U.S. Provisional Patent Application No.61/043,396 priority, the description and entire disclosure of this application are drawn herein It is used as reference.
Technical field
Present disclosure relates generally to biology, medical science and epidemiology field, in particular it relates to one Kind or it is various for diagnose, prevent and/or treat the mankind, animal, plant and other biological infectious disease, cancer, itself The method of immune disease, anaphylaxis, toxicity, obesity and/or other diseases.More specifically, the disclosure is related to And the method for being identified to the therapeutic target of above-mentioned disease, and resulting application.
Background technology
Saccharide is the important composition composition of life entity, can be alternatively arranged as stable as constituent and energy storage composition The factor (stabilization agent), recognition factor (recognition agent), signal factor (signaling Agent) and communication the factor (communication agent).It has recently been found that the saccharide of cell surface relates generally to adhesive attraction And the cell-cell interaction being thus related to, thus the interest of glycobiology is increasingly increased.Molecular biology is in this field In application, scientists is operable to the expression of saccharide and is studied the function of glycoprotein.
Difficulty in sugared structural research
The transmutability of sugared structure, partially due to monosaccharide units are be combined with each other in a number of different ways, this and protein Aminoacid or the nucleotide of DNA to be generally combined together in the standard fashion be contrary.The complexity of sugared structural research, also exists In its biosynthetic direct template is lacked, this is with the aminoacid sequence of protein by gene corresponding with them come the feelings for determining Condition is contrary.
Saccharide is also two grades of gene outcomes, and they are many enzyme synergism in subcellular fraction area by cell and produce 's.Therefore, the structure of saccharide possibly relies on expression, activity and the accessibility of different biosynthetic enzymes.This means not Substantial amounts of sugar may be produced using recombinant DNA technology as widely used in protein research, for structure and Functional study.
The content of the invention
Disclosure of the invention is based partially on following concept：The polysaccharide or saccharide of cell surface relates generally to intercellular mutual Effect；The polysaccharide or saccharide of at least some of form be some or it is all biological common in life starting and evolutionary process； And saccharide can change under different physiological statuss.Thus, the disclosure illustrates a kind of simple effective Based on polysaccharide or the method based on saccharide, for Rapid identification biomarker and therapeutic target, more particularly, to the mankind, Animal, plant and other biological infectious disease, cancer, autoimmune disease, anaphylaxis, inflammation, toxicity, obesity And/or the polysaccharide related target of other diseases.Existed according to the feature of the method in an embodiment of the disclosure of invention In following operation：
1) by the health and disease cells and/or tissue of organism, pathogen, polysaccharide, agglutinin, polysaccharide identifying system, Antibody and/or serum, medical herbs, small molecule and toxin (all these hereinafter referred to as target candidates), adhere to and/or are fixed on use On at least one solid carrier of chip (arry) or microchip (hereinafter referred to as chip carrier)；
2) make antibody or serum, pathogen, polysaccharide, agglutinin, polysaccharide identifying system, medical herbs, small molecule or toxin (all These are hereinafter referred to as " detection candidate ") combine on one or more chip carrier；
3) carry out to detecting candidate and the combination of the target candidate being located on one or more chip carrier Detection；
4) in zoopery and/or in cell or tissue culture systems, to be attached to one or more chip At least one target candidate on carrier have the biological function of the positive detection candidate for combining, pathogenesis, pharmacology, Toxicity is detected；
5) by step 4) in determine be related to infectious disease, autoimmune disease, anaphylaxis, cancer, obesity and The detection candidate of other diseases, in following application：Diagnose, prevent and treat these diseases；The exploitation and delivering of medicine； Vaccine development and epidemiology and biology (especially development and Evolutionary Biology) field；
6) to the therapeutic with healthy and/or diseased tissue and the cell of at least one detection organism for being combined of candidate Target spot or labelling (being attached to one or more chip carrier) are identified；
7) by step 6) in identification the therapeutic target or labelling, their derivant (including but not limited to：It is similar Thing, promote thing, antagonist, variant (variants), mutant (mutants), fragment, synthetic peptide, recombinant antigen) and it is any its The therapeutic target of its form, in following application：That is, with least one etiology and/or known to pathogenesis or unknown The related infectious disease of the therapeutic target, autoimmune disease, allergy, cancer, obesity and other diseases Application in diagnosis, prevention, treatment and medicine delivery.
Correspondingly, in an embodiment of the disclosure of invention, there is provided following methods：Control for Rapid identification The simple and efficient method of the property treated target spot；For drug development and the simple and efficient method of drug delivery system；For The research of pathogenesis and the mankind, animal, plant and other biological infectious diseases, autoimmune disease, anaphylaxis, The simple and efficient method of the cause of disease screening of toxicity, cancer, inflammation, obesity and other diseases；It is new for developing high-quality The simple and efficient method of type vaccine；For the simple and efficient method of effective control being very popular property disease；For coagulation The simple and efficient method that the function of element, medical herbs, toxin and small molecule, toxicity, pharmacology and pharmacy are studied；For developing certainly The simple and efficient method of the animal model of body immune disease, anaphylaxis, toxicity, cancer, obesity and other diseases；With And for studying the simple and efficient method of epidemiology and biology (especially Evolutionary Biology).By specific enforcement Mode, can more clearly understand various other purposes, feature and the beneficial effect of the present invention.
Description of the drawings
Figures of the Fig. 1 for display chip example.
The figure that Fig. 2 is combined with the tissue slice of adult rats for display phytohemagglutinin WGA and soybean agglutinin (SBA).
Fig. 3 is display phytohemagglutinin wheat germ agglutinin (WGA) and UEA I (UEA I) and healthy new lactogenesis The figure that the tissue slice of the small intestinal of Mus and adult rats combines.
Fig. 4 is for display phytohemagglutinin WGA and infection and is uninfected by rhesus rotavirus (rhesus rotavirus (RRV) figure that the tissue slice of neonatal rat) combines.
Fig. 5 is to show anti-RRV polyclonal antibodies and agglutinin WGA with RRV infection and the tissue slice of the neonatal rat being uninfected by The figure for combining.
Fig. 6 is the figure for showing the neonatal rat infected with the RRV of magistral medicines and brine treatment.
Specific embodiment
The disclosure is easily formed the embodiment of various multi-forms, herein, will be described in detail this The embodiment of the preferred implementation and replaceability of bright disclosure.It is to be understood, however, that the disclosure The example of the principle of the invention is considered, and the present invention and/or right to limit illustrated by embodiment will not The spirit and scope asked.
Chip carrier and attachment or/and fixed material and chemical reagent
In the present disclosure of the invention, for chip or microchip referring to can for solid carrier (hereinafter referred to as chip carrier) For aufwuch material and the object of chemical material, including but not limited to：Microscope slide (slide), flat board (plate), film, Bar (strip), chip or granule etc. are not limited.Biomaterial and chemical material can be affixed at least one chip and carry On body.For the method adhered to on solid carrier and fix biomaterial and chemical material, can be physical method, chemistry side Method, biological method and all other method known in the art.
For the material and chemical reagent of preliminary screening
According to disclosure, the biomaterial and chemical material include but is not limited to following material.
Organism and pathogen
In the present disclosure of the invention, term " organism " refers to independent biosystem, including but not limited to：Animal, Plant, insecticide, funguses or microorganism.Based on cell type, organism can be divided into protokaryon group and eucaryon group.Prokaryote is usual It is expressed as two different scopes, referred to as antibacterial and archeobacteria (Archaea).Eukaryote is included but is not limited to：It is the mankind, dynamic Thing, plant, funguses, Acarasiales (slime mould), algae, organelle, mitochondrion and (in plant) chromatoplast (plastids), viral eukaryotic origin (viral eukaryogenesis) etc..Recently, by Thomas Cavalier-Smith proposes a kind of Neomura clade, and archeobacteria and eukaryote are classified as a class.Cavalier- Smith also proposes that Neomura is evolved by antibacterial, is more specifically evolved by actinomycetes.
(microorganism (can also be spelled as micro organism) is microbe) a kind of with micro- for microorganism The visible organism of mirror (too little so that can not be seen by the bore hole of people).The disclosure of invention relates in one aspect to micro- life Thing, including but not limited to：The related life of the related pathogenic microbes of beneficial microbe, archeobacteria, disease and/or life concern Object.More specifically, microorganism is included but is not limited to：Antibacterial, virus, funguses, viroid, Protein viruss (prions) etc..
" pathogen " used herein refers to pathogenic organism, including but not limited to：Microorganism, parasite, insecticide, plant etc. Do not limit.Term " infectious disease " refers to that external source species are deleteriously invaded and grows in host organisms.Used by this method to infectious disease The pathogen that disease has specificity is included but is not limited to：Virus, antibacterial, parasite, funguses, viroid, Protein viruss, protozoacide And insecticide (protozoa).
The type of pathogen includes but is not limited to any type of pathogen, for example, living or dead or inactivation disease Substance, it is fresh or be dried pathogen, fixation or the pathogen of freezing, all or part of pathogen or fragment, cause of disease Body section, the homogenate of pathogen smear, pathogen, pathogen solute and pathogen extract etc. are not limited.
Antibody
Terminology used herein " antibody " refers to the immunocompetence portion of immunoglobulin molecules and immunoglobulin (Ig) molecule Point, i.e., the molecule of the antigen binding site containing specific binding (generation immunoreation) antigen.This antibody-like includes but does not limit In：Polyclonal antibody, monoclonal antibody, chimeric antibody, single-chain antibody, F.sub.ab, F.sub.ab ' and F (ab ') .sub.2 pieces Section and Fab expression libraries.Generally, it is related to IgG, IgM, IgA, IgE and IgD of any classification available from the antibody molecule of the mankind, this A little classifications are different each other by the property of the heavy chain being present in molecule.Specific classification also has subclass, for example IgG.sub.1, IgG.sub.2 etc..Additionally, in the mankind, heavy chain can be κ chains or λ chains.The antibody herein being related to includes institute These classifications of somebody's antibody-like kind, subclass, antibody fragment and type.In vertebrate blood or other body fluid, send out Existing naturally occurring antibody.
Be suitable for the present invention antibody can specifically resistant to arbitrary organism, any pathogen, any infectious agent, appoint Meaning polysaccharide, arbitrarily any saccharide complex (glycoconjugates), " autoantigen of organism or any biomarker " etc. Do not limit.Also include from be diagnosed as infectious disease, autoimmune disease, anaphylaxis, toxicity, cancer, obesity and other The serum of the patient of disease.
The antibody for being suitable for the antiviral of this method preferably includes, but is not limited to any type of anti-of anti-following virus Body or antibody fragment：Anti-double-chain DNA virus, single-stranded DNA viruses, diplornaviruses, (+) just strand rna virus, (-) antisense strand RNA viruses, RNA retrovirus retrovirus, DNA retrovirus retrovirus, associated viruss (satellites), hepatitis triangle virus (hepatitis delta virus), viroid, wherein, the double-stranded DNA viruses are included but is not limited to：Adenoviruss (adenoviridea), herpesviruss (herpesviridea), papovavirus (papovaviridea), poxvirus (poxviridea)；The single-stranded DNA viruses include but is not limited to：Orbiviruses (circoviridea), gemnivirus (geminiviridae), parvovirus (parvovirinae)；The diplornaviruses are included but is not limited to：Birnaviruss (biraaviridae), new virus (reoviridea)；(+) justice strand rna virus are included but is not limited to：Astroviridae (astroviridea), Caliciviridae (caliciviridea), coronaviridae (coronaviridea), flaviviridae (flaviviridea), picornaviridae (picomaviridea), marmor upsilon section (potyviridea), Ta Bamo Viraceae (tabamoviridea), tobacco mosaic virus (TMV) section (togaviridea)；(-) the antisense strand rna virus include but It is not limited to：Filamentous virus (filoviridea), Paramyxo viruss (paramyxoviridea), Pneumovirinae (pneumovirinae), rhabdoviruses (rhabdoviridea), shape of sand virus (arenavirus), Bunyavirus (bunyaviridea), orthomyxovirus (orthomyxoviridea)；The RNA retrovirus retrovirus are included but is not limited to：It is inverse Retroviral (retroviridea)；The DNA retrovirus retrovirus are included but is not limited to：Bacilliform DNA virus (badnavirus), Cauliflower mosaic viruses (caulimoviridea), hepadnaviruss (hepadnaviridea) associated viruss include but It is not limited to：Tobacco necrosis viruss associated viruss；The viroid includes but is not limited to：Potato spindle tuber viroid (potato spindle tuber viroid) and cause spongiform encephalopathy (spongiform encephalopathies) Virus.More specifically, any type of antibody of the including but not limited to anti-following virus of the antibody of antiviral：Reoviruses (reovirus), rotavirus (rotavirus), cytomegaloviruses (cytomegalovirus), influenza virus (include birdss A Type influenza virus), Epstein-Barr virus (Epstein-Barr virus), hepatitis viruss, the thermophilic T cell disease of HIV (human immunodeficiency virus) (HIV), the mankind Malicious (HTLV), human papillomavirus (papilloma virus), polioviruses (polio virus), parainfluenza viruses (parainfluenza virus), Measles viruss (measles virus), mumps viruss (mumps virus), respiratory tract Syncytial viruses (respiratory syncytial virus), parainfluenza type 3 virus (shipping fever virus), east type Encephalomyelitis virus and west type encephalomyelitis virus (Western and Eastern encephalomyelitis Virus), Japanese Type B encephalomyelitis virus (Japanese Bencephalomyelitis virus), Russian spring-summer brain Encephalitis virus (Russian spring-summerencephalomyelitis virus), swine fever virus (hog cholera Virus), poxvirus (pox virus), rabies viruss (rabies virus), distemper virus (distemper virus), Foot and mouth disease viruses (foot and mouth disease virus), rhinovirus (rhinovirus), Avian pneumo-encephalitis virus (Newcastle disease virus), vaccine viruses (vaccinia virus), and pseudorabies viruses (pseudorabies virus) etc. is not limited.
The antibacterial antibody for being suitable for this method preferably includes, but is not limited to resist the antibody of following any type antibacterial Or antibody fragment：Gram-positive and gram negative bacteria, or bacillus (shaft-like), coccus (spherical) and spirilla (curved wall shape) and other antibacterials.Specific antibacterial includes but is not limited to：Cholera bacteria (cholera), syphilis fungus (syphilis), anthrax (anthrax), leprosy (leprosy) and glandular plague bacterium (bubonic plague), rickettsia (rickettsias), Diplococcus gonorrhoeae (neisseriagonorrhoeae), bordetella pertussis (bordetella Pertussis), escherichia coli (escherichia coli), Salmonella (salmonella enterica), vibrio cholera (vibrio cholerae), Pseudomonas aeruginosas (pseudomonas aeruginosa), Yersinia pestis (yersinia pestis), Francisella tularensis (francisella tularensis), hemophilus influenza (haemophilus influenzae), purple sulfur bacteria (purple sulfur bacteria), helicobacter pylori (helicobacter pylori), campylobacter jejuni (campylobacter jejuni), Bacillus anthraciss/wax-like bud Spore bacillus/bacillus thuringiensiss (bacillus anthracis/cereus/thuringiensis), tetanus fusiform spore Bacillus (clostridium tetani), Clostridium botulinum (clostridium botulinum), staphylococcuses (staphylococci), streptococcus (streptococci), streptococcus pneumoniae (pneumococci), streptococcus pneumoniae (streptococcus pneumoniae), mycoplasma (mycoplasmas), bacteroides fragiliss (bacteroides Fragilis), Mycobacterium tuberculosiss (mycobacterium tuberculosis), Mycobacterium leprae (mycobacterium Leprae), diphtheria corynebacterium (corynebacterium diphtheriae), Tyreponema pallidum (treponema Pallidum), borrelia burgdorferi (borrelia burgdorferi), chlamydia trachomatiss (chlamydia Trachomatis), chlamydia psittaci (chlamydia psittaci), phycocyanin (phycocyanin), phycoerythrin (phycoerythrin), mitochondrion, chloroplast etc. are not limited.
The cell of organism, tissue and organ
Term " disease " used herein refers to the organism abnormal conditions of destruction body function, with special symptom and levying Million.In the mankind or animal, " disease " is generally more broadly used for referring to the discomfort for causing patient, dysfunction, pain, society Friendship problem (social problems) and/or any situation of death, or contact the class of those human or animals of the patient Like problem.In the broadest sense, disease sometimes include deformity, imbalance, syndrome, infectious disease, single symptom, Deviant Behavior, And structural and functional SARS form variation, and in other contents and for other purposes, these can be recognized To be diacritic category.The type of disease is included but is not limited to：The infection of the mankind, animal, plant and other biologies Property disease, cancer, autoimmune disease, anaphylaxis, toxicity, obesity and/or other diseases.
The cell of the Eukaryotic health or normal and disease of disclosure of the invention, tissue and/or device The type of official can be：Any type of cultured cell in vitro, including but not limited to：Various kinds of cell system well known in the art and original For cell；Any type of cell available from flesh tissue；Tissue that is any type of fresh, freezing or fixing or organ Fragment, section or smear；Cell or tissue or the extract or homogenate or solute of organ or any type of partial organ, Or the extract or homogenate or solute of any other type of cell, tissue or organ.
The mankind, the health of animal or plant and disease cells or tissue or organ include their partial or complete life Period, from embryo, fetus, new life, childhood until growing up.The health and unhealthy tissue of human and animal or organ can be (but not limited to)：Epithelial tissue and body of gland；Connective tissue；Muscle, including smooth muscle, skeletal muscle and cardiac muscle；Nervous tissue, including Central nervous system (CNS) and peripheral nervous system (PNS)；Cartilage, bone and joint；Extracellular matrix；Blood and hemopoietic system； Bone marrow；Cardiovascular system, including heart, tremulous pulse, blood capillary and vein；Respiratory system, including lung, bronchial tree, alveolar be little Pipe and alveolar；Digestive system, including oral cavity, esophagus, stomach, small intestinal (duodenum, jejunum and ileum) and large intestine (caecum, colon, Rectum, anal passage and vermiform appendix), salivary gland, pancreas, liver, bile duct and gallbladder；Urinary system, including kidney, ureter, bladder and urethra； Female repro ductive system, including ovary, fallopian tube, uterus and vagina；Male reproductive system, including testis, gonoduct, penis, essence Capsule, prostate and cowper gland；Lymph (immunity) system, including lymph node, thymus and spleen；Endocrine gland, including pinuies, hang down Body, thyroid, parathyroid gland and adrenal gland；Crust (integument), including skin and it appurtenance, sweat gland, sebaceous gland, Hair and fingernail.
Polysaccharide and saccharide complex
Term " polysaccharide " refers to polysaccharide or oligosaccharide.Oligosaccharide is the sugar containing peanut (typically 3-10) saccharic composition (monosaccharide) Birds of the same feather flock together compound.Polysaccharide is generally by monosaccharide by the amino acid side chain in O- or N- glycosidic bonds and suitable protein or fat part Connect and constitute.There is two kinds of glycosylation：The glycosylation of the N- connections being connected on the amide nitrogen of asparagine side chain, And the glycosylation of the O- connections being connected on the hydroxyl oxygen of serine and threonine side chains.Other polysaccharide include but do not limit In：O-GlcNAc, GAG chain, glycosaminoglycan (glycosaminoglycan), glycosyl sphingolipid (glycosphinglipid).Monosaccharide bag Include but be not limited to：Fructose, glucose, mannose, fucose, xylose, galactose, Lactose, N-acetylgalactosamine, N- acetyl Portugal Grapes glucosamine and sialic acid.The polysaccharide of O- connections and the polysaccharide of N- connections are common in eukaryote, although in prokaryote not It is common, but it is also possible to find.It can be found that polysaccharide is attached to protein, becomes glycoprotein (glycoproteins) or albumen is more Sugared (proteoglycans).They are usually located at the outer surface of cell.
Sialic acid is that the N- of the monosaccharide neuraminic acid (neuraminic acid) for describing 9 carbon atoms replaces or O- replaces Derivant generic term.The entitled N-acetyl-neuraminate (Neu5Ac or NANA) of the most common member in this group and 2- ketone generation -3- deoxidations octanoic acid (Kdn).Other members of sialic acid include but is not limited to：N-Acetyl-D-glucosamine (GlcNAc), N- Acetylgalactosamine (GalNAc), ManNAc (ManNAc) and NeuGc ALPHA2-3Gal (Neu5Gc).Sialic acid It is widely distributed in animal tissue and antibacterial, especially in glycoprotein and ganglioside.Amino group contain acetyl group or Glycolyl group.Hydroxyl substituent largely can be converted：What is had found has acetyl group, lactyl, methyl, sulfate Group and phosphate group.In the mankind and other biologies, the glycoprotein rich in sialic acid can be with reference to selection albumen (c-type coagulation Element).
Saccharide complex
In the present disclosure of the invention, term " polysaccharide " also refers to the carbohydrate fraction of saccharide complex, the saccharide complex bag Include but be not limited to：Glycoprotein, glycolipid, proteoglycan and glycophosphosphingolipid (glycophosphosphingolipids), or other Known or unknown saccharide complex.Saccharide complex is mainly found in outside cell wall and juice.Except saccharide complex itself, Due to there is the receptor that various polysaccharide are combined in cell surface, thus saccharide complex is critically important for intercellular interaction.http://en.wikipedia.org/wiki/Glvcobiology-note-Ma 2004
Term " proteoglycan " represents the glycoprotein of the high glycosylation of particular category.They include core protein and one Or multiple covalently bound glycosaminoglycan (GAG) chains.These glycosaminoglycan chains are long linear carbohydrate polymers, due to there is sulfur Acid and galacturonic acid groups, they carry negative charge in physiological conditions.Proteoglycan can by their glycosaminoglycan chain come Classification.These chains are included but is not limited to：Chondroitin sulfate (chondroitin sulfate) and dermatan sulfate (dermatan sulfate)；Heparin and Heparan sulfate；Keratan sulfate.Proteoglycan can also be classified by size.Big albumen The example of polysaccharide is collectin polysaccharide (aggrecan) and multifunctional protein polysaccharide (versican), and collectin polysaccharide is soft Major protein polysaccharide in bone, multifunctional protein polysaccharide are present in many adult tissues, and these adult tissues include but do not limit In blood vessel and skin.The little proteoglycan (SLRPs) repeated rich in leucine is included but is not limited to：Decorin (decorin), two polysaccharide (biglycan), fibromodulin (fibromodulin) and inner chamber albumen (lumican).
Term " glycolipid " refers to the lipid with reference to saccharide.Produce when phospholipid on expolasm surface of the sugar chain with cell membrane combines Raw glycolipid.They are expanded in extracellular aqueous environment from phospholipid bilayer, can as the recognition site of chemical compound specifically bind, and Help maintains the stability of film, while making cell be attached to each other to form tissue.Glycolipid is included but is not limited to：Galactolipid, sulfur Fat (SQDG), glycosphingolipid (glycosphingolipids), cerebroside (cerebrosides), galactocerebroside (galactocerebrosides), glucocerebroside (glucocerebrosides), glucoside (fat) ceramide (glucobicaranateoets), ganglioside (gangliosides), globoside (globosides), sulfur glycosides Fat (sulfatide), glycophosphosphingolipid (glycophosphosphingolipids) or any other known or unknown sugar Fat.
Polysaccharide identifying system
Polysaccharide it is assumed first to synthesize in the form of simple homotype polysaccharide (amylose, cellulose etc.), enters to turn to multiple Miscellaneous heterozygosis polysaccharide.This kind of evolution hypothesis has caused the appearance of the protein (agglutinin) for being related to " polysaccharide identifying system ", described Identifying system can recognize various structural molecule, identification molecule, can introduce bio signal and promote infectious disease.Polysaccharide The synthesis system and the identifying system interdepend, and think still carrying out coevolution.
Polysaccharide identifying system is included but is not limited to：Agglutinin (includes zoo-agglutinin, phytohemagglutinin and pathogen coagulation Element), the enzyme containing carbohydrate recognition domain (CRD), the antibody of anti-polysaccharide, the antibody of antibacterial agent, anti-chaperone it is anti- The antibody of body and anti-transport protein, the albumen with reference to Microbial saccharides, the albumen with reference to glycosaminoglycan or it is any other known to or Unknown polysaccharide identifying system.
Agglutinin is carbohydrate-binding protein, and their sugar moieties are high specials.In the present disclosure of the invention, coagulation Element is included but is not limited to：Zoo-agglutinin, phytohemagglutinin, pathogen agglutinin and any other known or unknown coagulation Element.Agglutinin is natural generally existing, typically containing the sugar identification domain guarded in evolution.Known agglutinin is by knowing Not in pathogen unique saccharide or on host cell inaccessible saccharide, and important work is played in immune system With.Pathogen agglutinin from virus, antibacterial, protozoacide and insecticide is active by their sialic acid identification, so as to relate to And infection.
Zoo-agglutinin is included but is not limited to：C-type agglutinin, M type agglutinins, L-type agglutinin, p-type agglutinin, R types coagulate Collection element, I type agglutinins, F type agglutinins, F box H type agglutinins (F-box H-type lectins), half lactadherin (galectins), penetrate plain (pentraxin), spider venom (spider toxin), Tyke agglutinin (tachylectin), Chitin binding protein, chitinase agglutinoid, TIM agglutinins, calnexin-calreticulin (calnexin- Calreticulin), Ficolins (ficolins), rich gram agglutinin (fucolectin), interior aggregation albumen And any other type of known or unknown zoo-agglutinin (intelectins).
Phytohemagglutinin is included but is not limited to：β-prism (prism) I type agglutinins, β-prism II type agglutinins, β-SANYE Careless agglutinin, the element (knottin) that knots, lectin of beans (legume lectin), Fructose specific agglutination element, mannose are special Different in nature agglutinin, glucose specificity agglutinin, fucose specific agglutination element, galactose specific agglutination element, N- acetyl half It is Lactose amine specific agglutination element and N-Acetyl-D-glucosamine specific agglutination element, and any other type of known or unknown Phytohemagglutinin.
Pathogen agglutinin is included but is not limited to：Bacterioagglutinin, viral agglutination element and mycoagglutinin.Bacterioagglutinin It is not limited to including seeing：AB5Toxin, antibacterial neurotoxin, staphylococcal entotoxin, cilium adhesin (pili adhensin), indigo plant are thin Bacterium agglutinin (cyanobacterial lectins), 1-Ca β-sandwich, 2-Ca β-sandwich, β-spiral, toxin repeat Domain.Viral agglutination element is included but is not limited to：Coat protein, hemagglutinin (hemmagglutinin), urogomphus play egg (tailspike protein), capsid spike protein and fiber spherical protuberances (fiber knob) in vain.Mycoagglutinin include but It is not limited to：Ig- sample agglutinins, actinoporin- sample agglutinins, β-Herba Trifolii Pratentis hole forms agglutinin, half lactadherin, 6- leaves Piece β-spiral and 7- blades β-spiral and any other type of known or unknown pathogen agglutinin.
Animal polysaccharide identification albumen is including but not limited to following two groups：Agglutinin and Sulfated glycosaminoglycan (SGAG) knot Hop protein.The biosynthesiss of baroque glycosaminoglycan (GAG) are adjusted, and define organ specificity and organizing specific Property and during g and D temporary various sulphation patterns.Passing through in addition to antibody and T-cell receptors The protein of immunoglobulin (Ig) class formation domain mediation polysaccharide identification is referred to as " I type agglutinins ".Combine with sialic acid (Sia) The main homologous subfamily of the I type agglutinins of characteristic and characteristic amino terminal architectural feature is referred to as " Siglecs " (identification Sia Ig superfamily agglutinins).
Mucin can be the glycoprotein containing sialic acid.Mucin is secreted in respiratory tract and gastral mucus.It is viscous Protein gene coding mucin monomer, the monomer synthesize shaft-like apomucin (apomucin) core, and by especially rich Rich glycosylation carries out post translational modification.Two zoness of different are found in ripe mucin：1) amino and carboxyl terminal area Domain, the region be it is slight glycosylated, but be rich in cysteine, may relate between the inside of mucin monomer and monomer Set up disulfide bond.2) big central area, is repeatedly formed by the series multiple of 10-80 residue sequence, in these residue sequences, The aminoacid of more than half is serine or threonine.The oligosaccharide institute saturation that the region can be connected by hundreds of O-.In viscous egg Can also find the oligosaccharide of N- connections in white, but negligible amounts.By cDNA clone, at least 19 people's mucoids are distinguished Gene-MUC1, MUC2, MUC 3A, MUC 3B, MUC 4, MUC 5AC, MUC 5B, MUC 6-9, MUC 11-13 and MUC 15- 19.The respiratory tract mucin of Major Secretory is MUC5AC and MUC5B, and MUC2 major parts are secreted in intestinal, but in respiratory tract In also have secretion.The mucinous generation increase in many adenocarcinoma, including cancer of pancreas, pulmonary carcinoma, breast carcinoma, ovarian cancer, colon cancer Etc..In pulmonary disease, mucin also has overexpression, such as asthma, bronchitis, chronic obstructive pulmonary disease (COPD) or capsule Property fibrosiss (cystic fibrosis).
Medical herbs and Chinese traditional herbs
Medical herbs used herein refer to the plant with value characteristic (such as medicinal property, fragrance, abnormal smells from the patient etc.).At this In bright disclosure, traditional Chinese herbal medicine includes but is not limited to Compendium of Materia Medica (unsimplified Hanzi：This Cao Gang mesh；Simplified Hanzi：This Careless detailed outline；Phonetic：běncǎo Gāngmù；Webster Pinyin method (Wade-Giles)：Pen-ts ' ao Kang-mu) (also referred to as Compendium of Materia Medica) in the medical herbs listed, Compendium of Materia Medica is write by the Chinese medicine that Ming Dynasty's Li Shizhen is write Make.It is a works for concluding Ming Dynasty's book on Chinese herbal medicine.List in the works and all of think the plant with medicinal property, dynamic Thing, mineral and other materials.
Especially, basic Chinese traditional herbs are included but is not limited to：Herba Pogostemoniss (Agastache rugosa), Radix Alangii (Alangium chinense), the Radix Pulsatillae (Anemone chinensis, Pulsatilla chinensis of the same name), Radix Anisodi Tangutici (Anisodus tanguticus), Herba Ardisiae Japonicae (Ardisia japonica), Radix Asteriss (Aster tataricus), the Radix Astragali or north Stilbene (Astragalus propinquus, Astragalus membranaceus of the same name), Camellia sinensis or Folium Camelliae sinensis (Camellia Sinensis), Fructus Cannabiss (Cannabis sativa), Flos Carthami (Carthamus tinctorius), Cortex Cinnamomi (Cinnamomum Cassia), Herba Cissampelotis or Asia slaves (Cissampelos pareira), short calyx Rhizoma Coptidis (Coptis chinensis), Rhizoma Corydalis (Corydalis ambigua), Fructus Crotonis (Croton tiglium), Flos Genkwa (Daphne genkwa), Flos Daturae (Datura Metel), datura (Datura stramonium, Datura tatula of the same name)[13], Herba Dendrobii or Herba Dendrobii (Dendrobium nobile), Radix Dichroae (Dichroa febrifuga)[14], Ephedra sinica (Ephedrasinica), the Cortex Eucommiae (Eucommia ulmoides), the Radix Euphorbiae Pekinensis (Euphorbia pekinensis)[15], a leaf autumn (Flueggea Sufjruticosa, once name Securinega suffruticosa), Fructus Forsythiae (Forsythia suspensa), Herba Violae It is (Gentiana loureiroi), Fructus Gleditsia (Gleditsia sinensis), Radix Glycyrrhizae (Glycyrrhiza uralensis), big Wind (Hydnocarpus anthelmintics, H.anthelminthica of the same name), Ilicis Purpureae (Ilex purpurea), Herba Leonuri Careless (Leonurus japonicus), Rhizoma Chuanxiong (Ligusticum wallichii)[18], Herba Lobeliae Chinensis (Lobelia Chinensis), Cortex Phellodendri (Phellodendron amurense), Cacumen Platycladi (Platycladus orientalis, once name Thuja Orientalis), golden larch (Pseudolarix amabilis), Herba Psilopegani sinensises (Psilopeganum sinense), Radix Puerariae (Pueraria lobata), snakewood or From snakewoods or India's letterwood (Rauwolfia serpentina), Radix Rehmanniae or dry ground Yellow (Rehmannia glutinosa), Rheum officinale (Rheum officinale), tsinghai rhododendron herb (Rhododendron Tsinghaiense), Saussurea lappa Clarke (Saussurea costus), Fructus Schisandrae Chinensis (Schisandra chinensis), Radix Scutellariae (Scutellaria baicalensis), the Radix Stemonae (Stemona tuberosa), Radix Stephaniae Tetrandrae (Stephania tetrandra), Chinese scholartree Or Sophora japonica L. or Flos Sophorae (Styphnolobium japonicum, once name Sophora japonica), Fructus Trichosanthiss (Trichosanthes Kirilowii), Radix Wikstroemae (Wikstroemia indica), Radix Isatidis (Isatis indigotica), YUNNAN BAIYAO, Radix Salviae Miltiorrhizae (Eclipta prostrate herb), Herba Taraxaci (Taraxacum mongolicum herb), Radix Ginseng (Ginseng), Radix Rehmanniae Preparata Yellow (Rehmannia glutinosa/foxglove root prep.), Herb Gynostemmae Pentaphylli (Panto Teapills), Rhizoma Dioscoreae (Dioscorea opposite/Chinese yam rhizome), Cortex Moutan (Paeonia suffruticosa/peony Tree root-bark), Poria (Poria cocos fungus/mushroom filaments), Rhizoma Alismatis (Alisma Plantago aquatica/water plantain rhizome), Fructus Corni (Cornus officinalis/dogwood Tree fruit), Cortex Cinnamomi (Cinnamomum cassia/cinnamon bark), Radix Aconiti Lateralis Preparata (Aconitum carmichaeli Root prep.), Radix Codonopsis (Codonopsis root), Radix Et Caulis Acanthopanacis Senticosi (Eleuthro root), Cordyceps (Cordyceps), Ganoderma (Reshi/Mushroom of Immortality), Radix Polygoni Multiflori (Polygonum multiflorum root), Semen Coiciss (Coix lachrymal jobi/Seeds of Jobs Ears seed), Ramulus Cinnamomi (Cinnamomum cassia/cinnamon Twig), Rhizoma Zingiberis Recenss (Zingiber officinalrhizome.Ginger root), the Radix Paeoniae Alba (Paeonia lactiflora/ White peony root), Radix Angelicae Sinensis (Angelica sinensis root), Radix Saposhnikoviae (Ledebouriella divaricata Root), Poria (Pori α cocos fungus), the Cortex Eucommiae (Eucommia ulmoides bark), Rhizoma Atractylodis (Atractylodes Lancea rhizome e), Radix Platycodoniss (Platycodon grandiflorum/ballon flower root), Olibanum (Boswellia carterii), Myrrha (Commiphora myrrha/myrrh resin), Rhizoma Corydalis (Corydalis Yanhusuo/fumewort rhizome), Semen Persicae (Prunus persica/peach seed), Cornu Cervi Pantotrichum (Deer antler), The Rhizoma Atractylodis Macrocephalae (Atractylodes macrocephala), Herba Menthae (Mentha halocalyx/field mint hearb), Radix Bupleuri (Bupleurum chinense root), Fructus Forsythiae (Forsythisia suspense fruit), the Radix Angelicae Dahuricae (Angelica Dahurica root), Pericarpium Citri Reticulatae (Citrus reticulate/citrus peel), Fructus Jujubae (Ziziphus jujube/ Chinese date fruit), Flos Chrysanthemi (Chrysanthemum morifolium flower), Semen Ziziphi Spinosae (Ziziphus Spinosa/sour jujube seed), Rhizoma Dioscoreae (Dioscorea opposite/Chinese yam rhizome), Semen Fagopyri Esculenti (Buckwheat), the Rhizoma Pinelliae (Pinellia ternata rhizome).
Inorganic ionss and small molecule
Inorganic ionss in the disclosure of invention include mineral nutrient, including but not limited to：Boron element, copper coin Element, manganese element, zinc element, molybdenum element, element sulphur, ferrum element, calcium constituent, potassium element, nitrate, phosphate, chloride etc. Do not limit.
Small molecule in the disclosure of invention is included but is not limited to：With reference to the peptide of polysaccharide, sugar chain (also referred to as aliphatic Sugar, and with common chemical formula CnHn+2)；Still there is the constitutional isomer of different structure with identical hydrocarbon chemical formula；No Saturated hydrocarbons, wherein, sugar chain contains multiple bond；Alcohol (the aliphatic carbon of one or more hydroxyls that carrying is directly connected with carbon atom Compound)；Aldehyde (secondary alcohols of ketone)；Organic acid (is produced by aldehyde radical oxidation)；Ester (is produced by the condensation reaction of alcohol and acid)；Or Any other known or unknown small molecule.
Toxin
Toxin in the disclosure of invention is included but is not limited to：Melittin (apitoxin), extracellular toxin (exotoxin), Endotoxin (endotoxins), cyanophycean toxin (cyanotoxins), necrotoxin (necrotoxins), hemotoxin (hemotoxin), mycotoxin (mycotoxin), neurotoxin (neurotoxin), light toxin (phototoxin), toxicity Group (toxicophore), toxoid (toxoid), venom (venom), Ricin (ricinis), or it is any other known or Unknown toxin.
The toxin is the noxious substance produced by cell living or organism, is active in low-down concentration.Poison Element can be small molecule, peptide or protein, and can when body tissue is contacted or when being absorbed by body tissue, due to biology Macromole (such as enzyme or cell receptor) interacts and causes disease.The order of severity of toxin is very different, generally from Less and acute (such as bee sting) is to almost fatal immediately (such as Botulinum toxin).Biotoxin is in purpose and mechanism Aspect is very different, and can be that (venom of cone shell (cone snail) contains tens kinds of little albumens to high complexity Matter, the nervous pathway or receptor of respective site-specific) or relatively small protein.
Primary screener
The material and chemical reagent for being used for primary screener as above, can serve as the target reagent of this primary screener method Or detectable.Hereinafter they are referred to as " target candidate " or " detection candidate ".
Target candidate
As described above, target candidate can be attached or fixed on carrier with multiple combination as needed.The target candidate Those listed in the combination of thing and detection candidate including but not limited to table 1.
The combination that 1 target candidate of table and detection candidate are combined
Note：Polysaccharide includes polysaccharide and polysaccharide complex.GRS=polysaccharide identifying systems.
In table 1, the tissue includes but is not limited to various health and the cell and tissue of disease of organism, medical herbs bag The Chinese traditional herbs for including but being not limited to the described above.Labelling "+" represents that two kinds of candidates are combined.
I types：Target candidate includes：Polysaccharide, pathogen and health tissues.Polysaccharide is at least included but is not limited to：Fructose, Fructus Vitis viniferae Sugared (Glc), mannose (Man), fucose (Fuc), xylose (Xyl), galactose (Gal), Lactose, glucamine (GlcN), half Lactose amine (GalN), mannosamine (ManN), N-Acetyl-D-glucosamine (GlcNAc), N-acetylgalactosamine (GalNAc), N- Acetylmannosamine (ManNAc), N-acetyl-neuraminate (Neu5Ac), NeuGc ALPHA2-3Gal (Neu5Gc), 2- ketone generation- 3- deoxidations nononic acid (Kdn), glucuronic acid (GlcA), galacturonic acid (GalA), mannuronic acid (ManA), and Iduronic acid (IdoA).Health tissues are included but is not limited to：From the various kinds of cell or tissue of at least one healthy organism, The healthy organism is included but is not limited to：The mankind, animal, plant or other biological body.
The detection candidate for being incorporated into I type target candidates is included but is not limited to：Polysaccharide, antibody, agglutinin are (solidifying including plant Collection element or other polysaccharide identifying systems), medical herbs, small molecule or toxin, according to need depending on.
II types：Target candidate includes：Polysaccharide (as described in I types), pathogen and diseased tissue.Diseased tissue is included but not It is limited to：From the various kinds of cell or tissue of at least one organism, the organism suffers from infectious disease, cancer, autoimmune At least one in property disease, anaphylaxis, toxicity, obesity or other diseases.Detection candidate with it is identical described in I types.
Type III；Target candidate includes：Polysaccharide, pathogen, health and diseased tissue (as described in I types and II types).Detection Candidate with it is identical described in I types.
IV types：Target candidate includes：At least one in below：Including but not limited to antibody, agglutinin are (solidifying including plant Collection element or other polysaccharide identifying systems), medical herbs, small molecule and toxin (depending on according to need).Detection candidate is included but is not limited to： Polysaccharide；The extract of the cell or tissue or organ of the health or disease of pathogen and organism, homogenate or solute, according to needing Depending on wanting.
V-type：Target candidate includes any two kinds such as the target candidate described in IV types.Described in detection candidate and IV types It is identical.
VI types：Target candidate includes any three kinds such as the target candidate described in IV types.Detection candidate and institute in IV types That what is stated is identical.
VII types：Target candidate includes any four such as the target candidate described in IV types.Detection candidate and institute in IV types That what is stated is identical.
VIII types：Target candidate includes：Antibody, agglutinin (including phytohemagglutinin or other polysaccharide identifying systems), grass Medicine, small molecule and toxin.Detection candidate with it is identical described in IV types.
Present invention additionally comprises target candidate and detection candidate according to need depending on other combination.
The combination of detection candidate and target candidate
The one side of the disclosure of invention is related to make detection candidate and carries at least one above-mentioned target candidate group The chip carrier of conjunction combines.The detection candidate can be pure or (such as biotin, fluorescence are appointed with conjugation moiety Anticipate other detectable things known in the art).If desired, the detection candidate being known in the art/carrier-bound various sides In method, it is possible to use the second reagent or the 3rd reagent.
Detection candidate is including but not limited to as described below with the binding pattern of the target candidate on chip carrier.
I-III types are tested
I types, II types and type III chip are the useful tools for the antigen in a kind of organism of Rapid identification, the life At least another kind of organism (especially pathogenic organisms or infectants) antigen of object simulation；And detection therapeutic target Property.Example includes：
A. it is incorporated into the polysaccharide of at least one pathogen：The polysaccharide is the possible binding site of the pathogen, and is appointed Meaning, is may each be as the health and/or diseased tissue of the organism of a part for cell or tissue structure with the polysaccharide The target spot of the pathogen.The pathogen may cause the related disease of these tissues or organ.
B. it is incorporated into the polysaccharide of multiple pathogens：The pathogen and the polysaccharide be for binding site vaccine, receptor/ The candidate of part vaccine or anti-multiple pathogens vaccine.
C. it is incorporated into the detection candidate of at least one pathogen and at least one organism health and/or diseased tissue： The pathogen and tissue share a therapeutic target.
If the detection candidate is antiviral antibody, the pathogen and the antibody may cause itself The Other diseases of the related biological injury of immune disease, anaphylaxis, toxicity or target tissue or organ.The antibody be with The candidate of lower application：For the diagnosis of associated conditions, antibody prevention and treatment, drug delivery machinery.For example, anti-rotavirus (RV) antibodies are in N-ACETYL-D-GLUCOSAMINE, and N-ACETYL-D-GLUCOSAMINE is in the heart of healthy mice, lung and small intestinal Expression (Fig. 2 and Fig. 3), and the expression in the cell (Fig. 5) of inflammatory cells (Fig. 4) or propagation.Therefore, anti-RV antibody can To be inflammation-induced agent, so as to cause the autoimmune disease or cancer of these tissues or organ.
If the detection candidate is polysaccharide, phytohemagglutinin (i.e. with reference to the agglutinin of mannose) or other polysaccharide known Other system, medical herbs, small molecule or toxin, then it is described to detect that candidate cause autoimmune disease, anaphylaxis, toxicity phase The biological injury or target tissue or the Other diseases of organ of pass；Or the detection candidate can be used for associated conditions The possibility candidate of diagnosis, prevention, treatment, medicine and drug delivery machinery.For example, specific recognition N- acetyl-D-Glucose The wheat germ agglutinin (WGA) of amine can cause the disease of the tissue or organ of expression N-ACETYL-D-GLUCOSAMINE.
D. at least one pathogen, the health of at least one organism and/or diseased tissue are incorporated into and at least one is more The candidate of sugar：The polysaccharide is the possible epi-position of the therapeutic target that the pathogen and the tissue are shared.The polysaccharide The disease related to the therapeutic target may be caused to its derivant, or the diagnosis as associated conditions, prevented, controlled The possibility candidate for the treatment of, medicine and drug delivery machinery.
IV-VIII cake cores
IV types, V-type, IV types, VII types and VIII cake cores are related to for diagnosing, preventing and treating for rapid screening The useful tool of the cause of disease, medicine and drug delivery machinery of the disease of therapeutic target and/or pathogen.Example includes：
A. combine the target candidate of the IV-VIII cake cores of polysaccharide：The target candidate may cause using polysaccharide as table The related disease of the therapeutic target of position；And the target candidate can be used for examining for the therapeutic target associated conditions Disconnected, prevention, treatment, medicine and drug delivery machinery.
B. it is incorporated into the target candidate of the IV-VIII cake cores of pathogen (i.e. virus)：The target candidate is for institute State the possibility candidate of diagnosis, prevention, treatment, medicine and the drug delivery machinery of the related disease of pathogen.
C. combine with the extract or homogenate or solute of the health or disease cells or tissue or organ of organism The target candidate of IV-VIII cake cores：The target candidate is for described biological cell or tissue or organ correlation The possibility candidate of the diagnosis of disease, prevention, treatment, medicine and drug delivery machinery.
D. (that is, anti-rsv antibodies and health influenza virus are anti-to be incorporated into the antibody of at least two different anti-different pathogens Body) polysaccharide：The pathogen is the candidate of binding site vaccine or multiple pathogens vaccine.
E. combine the pathogen of antibody (i.e. anti-rsv antibodies and antibodies against influenza virus) of various anti-different pathogens (i.e. RSV)：The pathogen is the candidate of binding site vaccine or receptor/ligand vaccine or multiple pathogens vaccine.
Present invention additionally comprises other chips of the combination containing other the target candidates depending on according to need and detection candidate.
The purification of therapeutic target and identification
Biological targets in one embodiment are biological ployose, such as protein or nucleic acid, and their activity can Regulate and control with by outside stimuluss.This definition depends on context, can refer to active pharmacological agent compound biological targets, Or the receptor target of hormone (such as insulin).One molecule is by signal hit and thus its behavior changes for hint.It is biological It is modal protein to learn target, such as enzyme, ion channel and receptor.Term " biological targets " is generally used for study of pharmacy, uses To describe the native protein of body, their activity is regulated and controled by medicine, with the therapeutic effect needed for obtaining.Herein In, the biological targets are often referred to drug target.Biomarker is used as the material of biological condition indicator.It is characterized in that Normal biological process, pathogenic course or anti-to the pharmacology of interventional are objectively measured and are evaluated as indicator Should.In the present disclosure of the invention, the therapeutic target for being related to organism disease is included but is not limited to：Biological targets, medicine Target, biomarker and pathogen binding site.
In another embodiment, include using choosing for identifying the straightforward procedure of functionally important therapeutic target The candidate (that is, antibody, preferably antiviral monoclonal antibody) of selecting property.The method can be saved in protein purification field Numerous and diverse work of conventional use of purpose antigen screening operation.Disclosure of the invention, the mankind as above, animal Or the available relevant cell of plant, the serosity of tissue and/or organ, solute or extract, can be used for general with this area Purification of the various ways known to logical technical staff to therapeutic target.
Also include with various ways well known to those of ordinary skill in the art, to therapeutic in the disclosure of invention The combination of the sequence or structure of target spot and therapeutic target and given candidate related main molecules, therapeutic target spread out Biology is identified.
Can be but be not restricted to according to the therapeutic target of the disclosure of invention：Glycoprotein；Polysaccharide (glycan)；It is many Peptide；Polysaccharide (polysaccharides)；Oligosaccharide；Lipid, glycolipid；Saccharide；Agglutinin, selection albumen；Mucin；Hemagglmination Collection element, collagen, Keratin, receptor (including virus receptor, toll sample receptors)；Cell component；Oncoprotein；Mammal is thin The fragment of born of the same parents' (including tumor cell), or any other material.
It is characterized in that according to the therapeutic target of an embodiment of the disclosure of invention, is depended on described herein The feature of target, therapeutic target can be preventative or pathogenic.
Cell or tissue culture assays and zoopery
In one embodiment, operations described below can be carried out to the candidate that positive combination is shown in primary screener.
Cell or tissue culture assays
According to an embodiment of the disclosure of invention, cell or tissue culture assays may be used to determine selected time Select the functional character and pathogenetic feature of the combination of thing or at least two selected candidates.I.e. target candidate or detection candidate are The no binding site for pathogen, or target candidate or detection candidate whether can induce significant biological conditions.Cell Or tissue culture's analysis can be also used for determining selected candidate or at least two times determined by the primary screener of chip Select pharmacology, kinetics and the toxic action of the combination of thing.Example includes：
A. the cell line sensitive to infectious pathogen known in the art, and primary healthy cell is organized or device Official's (i.e. target of disease of viral infection), can select the combination of candidate with the selected candidate of various dosage or at least two Together culture a period of time, the time make the candidate enough be present in the cell or tissue or organ on the candidate The targeting of thing combines, and should wash away the free candidate of uncombined target, processes described thin with the pathogen (i.e. Strain) For a period of time, the time makes the pathogen enter the target cell enough for born of the same parents or tissue or organ.Can be general by this area Various methods known to logical technical staff (determine the Strain drop in culture supernatant come the infection for detecting the pathogen Degree).
Selectively, same cell or tissue culture systems can be cultivated together with the infectious pathogen of given dose For a period of time, the time make the pathogen enter the target cell enough, remaining free cause of disease in culture fluid should be washed away Body, and the selected candidate of the cell or tissue or organ and various dosage or at least two are selected the combination of candidates Cultivate together.The infection of the pathogen is detected as described above can.
In the case where the target is the binding site of the pathogen, or the target is to be related to entering for the pathogen In the case of the factor for entering or infecting, the selected candidate or at least two combinations for selecting candidates can block described Target, and prevent the pathogen from entering or infecting the cell or tissue or organ.Thus, before and after infection, with institute State selected candidate or at least two select candidates combined treatment the cell or tissue or organ, will not infect or Pathogen described in mild infection.The method can be also used for the quick exploitation medicine related to infectious disease and drug delivery system System.
B. same cell as above or culture systems are prevented, can individually with the selected candidate of various dosage or A period of time is cultivated at least two combinations for selecting candidate together.The combination of candidate and at least two candidates increases to cell Grow, signal transduction, apoptosis, the effect of other functions of necrosis and cell or tissue or organ, can be common by this area Various methods known to technical staff are detecting.
The method can be used for the functional study and pathogenesis of at least two combinations for selecting candidate, and The cause of disease for autoimmune disease, anaphylaxis, inflammation, toxicity, cancer, obesity and other diseases is screened；For selecting The toxicity of candidate, pharmacology and medicinal study；For the quick exploitation medicine based on target spot related to those diseases and medicine Thing delivery system.
C. disease cells system (i.e. the cell line derived from tumor tissues) known in the art, and primary disease cells Or tissue or organ (i.e. tumor cell or tissue), candidate can be selected with the selected candidate of various dosage or at least two Combination cultivate together a period of time.The combination cell proliferation of candidate and at least two candidates, signal transduction, apoptosis, Necrosis, and disease cells or tissue or organ other functions effect, can by those of ordinary skill in the art it is ripe The various methods known are detecting.
The method can be used for the pathogenesis of the combination for selecting candidate or at least two selected candidates, and The cause of disease for autoimmune disease, anaphylaxis, cancer, inflammation, obesity and other diseases is screened；For selecting candidate Or at least two toxicity of combination, pharmacology and medicinal studies for selecting candidate；It is related to those diseases for quick exploitation The medicine based on target spot and drug delivery system.
Zoopery
Another theme of the disclosure of invention is zoopery it is determined that selecting target or at least two selected candidates Combination functional characteristic and pathogenic features in application.I.e. whether therapeutic target is the binding site of pathogen, or Whether therapeutic target induces significant biological conditions in organism.Animal (i.e. embryo, the tire at various ages can be used Youngster, new life, childhood and grow up) assessing in embryo or period of fetus strongly expressed as growth table is up to the therapeutic target for reducing Feature.
The zoopery can be also used for determining candidate or at least two candidates determined by the primary screener The pharmacology of the combination of thing and toxic action, and for quick exploitation and infectious disease, autoimmune disease, allergy Disease, cancer, inflammation, toxicity, the obesity medicine based on target spot related to other diseases and drug delivery system.
The symptoms such as disease, the death of related to animal are checked daily.It is determined that time point (i.e. process after 1 day, 3 My god, 5 days, 7 days, 14 days, 21 days, 28 days), collect the blood and target of the animal, and control organ or tissue (in primary sieve Choose determination).By various methods (i.e. Histological change) well known to those of ordinary skill in the art, with the sample of the collection Product to inflammation, pathogenic, toxicity, cell propagation, signal transduction, wither assessing the combination of candidate or at least two candidates Die, necrosis, and the animal target tissue or organ other functions effect.The animal that will can also be processed with candidate It is compared with the control animal of unused candidate process, to detect pathogen and/or candidate with target tissue and/or target organ Combination.Example includes：
A. selected candidate or at least two can be selected the combined administration of candidates when one section of the animal (i.e. mice) Between, the time makes the candidate or the combination of at least two candidates combine with the target of the animal or phase interaction enough With.Then the animal is made to be subject to the attack of pathogen (i.e. virus).It is related to entering for the pathogen in the target In the case of binding site or the factor, the combination of the candidate or at least two candidates can be blocked or affect the combination Site, and the pathogen is prevented from into the target cell of the animal.Thus, the animal will not infect or slightly feel Contaminate the pathogen.
Identical animal model can be used, by selecting candidate or at least with rear administration to animal in following pathogen challenge The combination of two kinds of candidates, determines therapeutic of the combination of the selected candidate or at least two candidates to relevant disease Effect.Those of ordinary skill in the art can be dynamic to process with the combination of the candidate of low dosage or at least two candidates Thing (such as mice), the scope of the low dosage can be produced and be effectively blocked and not excessive；Connecing for virus can be carried out at second day Kind.Apply the combination for selecting candidate or at least two candidates in can also be after virus inoculation one day.The common skill in this area Art personnel can carry out remaining experiment, including symptom, the measure virus titer of assessment infection.
B. selectively, the combined treatment phase of candidate can be selected with the selected candidate of various dosage or at least two Same animal.Selected candidate or at least two combinations for selecting candidate can be applied by intravenouss administration, intraperitoneal, muscle Interior administration, subcutaneous administration, intracavity administration, applied dermally, suction are applied, or are otherwise applied.Selected candidate or at least The combination of two kinds of selected candidates can be used to animal pregnancy, and is detected in the newborn or growing animal that the mother gives birth to.
If selecting being administered alone for the combination of candidate or at least two selected candidates, can induce in animal significantly Biological conditions, then be pathogen by the target and the candidate identification, the animal can serve as the examination of the disease Test model.
C. can use and be collected from, with the tissue of the animal of candidate individual processing, detecting candidate in vivo and cell Or the combination of tissue or organ.The available combination candidate in the disclosure of invention, it is also possible to for in-vivo imaging, its In, for example, the mankind or animal will be applied to the selected candidate of detectable part labelling, be preferably applied in blood flow, and examine Survey the presence of the candidate of labelling and positioning described in the host.The candidate be able to can be examined in being used in the mankind and/or animal The arbitrary portion of survey carrys out labelling, can be examined by nuclear magnetic resonance, NMR, lonizing radiation, fluorescence or other detection meanss known in the art Survey.
D. further preferably the combination with the internal and external method of detection method is combined using candidate in animal.For example, Selected candidate with mark part can be applied to animal, the candidate can be combined with its target or mutual in vivo Then the animal is cut by effect, collects tissue or organ samples, and in vitro with detection meanss known in the art examining Survey the candidate for combining.
E. system embryo culture
Candidate can be detected to pathogen (i.e. virus) using organism embryo (i.e. chicken or Brachydanio rerio) culture systems Inhibitory action.Those of ordinary skill in the art can be selected using the selected candidate of at least one of low dosage or at least two The combination of candidate processing mid-stage embryos, the dosage produce effectively blocking or act on and in not excessive scope；Can be Carry out pathogen inoculation within second day.Candidate can also be applied within one day after pathogen inoculation.Ordinary skill people Member can carry out remaining experiment, including collect the liquid containing pathogen, detection pathogen titre.With candidate or at least two A kind of combined treatment of candidate embryo culture thing with not with candidate process matched group compared with, with relatively low cause of disease Body titre, represents with effectively prevention or therapeutical effect.
Present invention additionally comprises other cell or tissue cultures on demand and zoopery.
The characteristic or feature of therapeutic target
Based on above-mentioned cell or tissue culture and zooperal result, the characteristic or feature and application bag of therapeutic target Include but be not limited to：
A types：Therapeutic target is the binding site of pathogen, rather than (therapeutic target will not in pathogenic site Notable biological conditions are induced in organism).
The therapeutic target can be by resisting that effective vaccine and the effective passive immunity without notable side effect are induced The receptor of body, and should be the target of vaccine development and antibody prevention and treatment.The therapeutic target or binding site The good immunogen candidate of vaccine.The accelerator of the therapeutic target and other derivants are for related to the target Disease (i.e. virus infection) prevention, treatment and drug delivery machinery good candidate.
The candidate that can be combined with therapeutic target as above, can be directly used as the pre- of associated conditions Anti-, treatment and the medicine of drug delivery machinery.
Type B：Therapeutic target is that (therapeutic target is in vivo for the binding site of pathogen and pathogenic site Induce significant biological conditions).
Such therapeutic target can be induced by effective passive immunity of effective vaccine or no notable side effect The receptor of antibody.The vaccine and antibody prevention and treatment safely should be applied.That is, when seeking having for pathogen binding site During effect blocking, it is intended to make any extra free antibodies reach minimum.The antagonist of the therapeutic target and other derivants It is the good candidate of the prevention, treatment and drug delivery machinery for the disease (i.e. virus infection) related to antigen.Can The candidate combined with therapeutic target as above, can cause inflammation, autoimmune disease, anaphylaxis, cancer, fertilizer Fat disease and other diseases.These candidates can serve as the medicine delivery of diagnosis, treatment and associated conditions for these diseases Instrument, can be applied with low dosage or be combined administration with the antagonist or inhibitor of the therapeutic target.These candidates are also Can be used for developing the animal model of associated conditions.
C-type：Therapeutic target is not the binding site of pathogen, but pathogenic site (the significant side effect of induction, poison Property or disease).
The therapeutic target using the type should be avoided in vaccine development.
The candidate that can be combined with the type therapeutic target can be the inducement of bio-toxicity, and have high mortality Disease (i.e. bird flu and ebola virus infection), autoimmune disease, anaphylaxis, cancer, inflammation, obesity and other The pathogenesis of disease.These candidates can be used for the diagnosis of these diseases, should avoid directly being treated with these candidates These diseases.However, these candidates can be used in combination with the antagonist of the therapeutic target or inhibitor, and with effect In the medicine and drug delivery machinery of associated conditions.On the other hand, these candidates can be used for developing associated conditions Animal model.
D types：Target spot neither pathogen binding site, nor pathogenic sites.
The therapeutic target of the type can cause the invalid of vaccine, should avoid for vaccine development.Can be with this type All applications that the candidate that therapeutic target is combined should avoid for organism, but if such combination be organ and/ Or it is tissue-specific, then the candidate can serve as drug delivery machinery.
For preventing and treating the realization of the new treatment based on binding site of infection, following mechanism can be based on：1) Compete with the pathogen binding site.2) block the pathogen binding site.3) modify the pathogen binding site Chemical characteristic.
For preventing and treating infection, cancer, autoimmune disease, anaphylaxis, inflammation, obesity and other diseases Based on the realization of the new treatment of pathogenic sites, based on following mechanism：1) pathogenic sites are suppressed；2) with the pathogenic induction In thing and/or compete；And 3) modify the chemical characteristic of the pathogenic sites.
Detailed description is easier to understand can various other characteristics or feature of therapeutic target.
Purposes based on the chip of polysaccharide
According to the disclosure of invention, chip based on polysaccharide and by the therapeutic target determined based on the chip of polysaccharide, With several purposes described herein, including the mankind, animal, plant and other biological purposes are suitable for, for example：1) it is quick to reflect Determine therapeutic target；2) infectious disease, autoimmune disease, anaphylaxis, toxicity, cancer, obesity and other diseases Pathogenesis and cause of disease screening；3) autoimmune disease, anaphylaxis, toxicity, cancer, obesity and other diseases are developed Animal model；4) develop the novel vaccine (vaccine i.e. based on binding site) of high-quality；5) effective control of the disease that is very popular (prevention and treatment i.e. based on binding site)；6) functional study of phytohemagglutinin, medical herbs, toxin and small molecule, toxicity are ground Study carefully, pharmaceutical research and pharmacy are studied；7) the quick exploitation of the medicine related to therapeutic target and drug delivery system；8) control Treat property target spot, their derivant and the therapeutic target any other form, with least one therapeutic target phase The diagnosis of the infectious disease of pass, autoimmune disease, anaphylaxis, cancer, obesity and other diseases, prevention, treatment and Application in medicine delivery；9) research of epidemiology and biology (especially Evolutionary Biology)；And 10) it is disclosed this Bright detailed description and PCT/US2007/018258, are easier to understand can many other purposes of molecular mimicry chip.
The related therapeutic target of polysaccharide and infectious disease
In an embodiment of the disclosure of invention, there is provided for understanding institute in PCT/US2007/018258 The etiology of the infectious disease stated, pathogenesis, the useful tool for treating and preventing.
Furthermore it is possible to develop the small animal model of HIV based on the new organ tropism (i.e. mice intestinal) of HIV. Toy includes but is not limited to mice, rat, Cavia porcelluss, rabbit etc..With great-hearted or inactivation HIV animal, Ran Houjian Survey the HIV tended in organ (i.e. mice intestinal).
The related therapeutic target of polysaccharide and inflammation, autoimmune disease and anaphylaxis
In another embodiment of the disclosure of invention, there is provided for understanding in PCT/US2007/018258 The etiology of described autoimmune disease, pathogenesis, the useful tool for treating and preventing.
In addition, with the similar approach as described in PCT/US2007/018258, antibody described in the disclosure of invention and its Its candidate can be used for inflammation, cancer, autoimmune disease and hypersensitive diagnosis, prevention and treatment.
The related therapeutic target of polysaccharide and cancer, obesity and other diseases
In the further embodiment of the disclosure of invention, there is provided for understanding in PCT/US2007/018258 Described cancer, the etiology of obesity and other diseases, pathogenesis, the useful tool for treating and preventing.
In addition, with the similar approach as described in PCT/US2007/018258, antibody described in the disclosure of invention and its Its candidate can be used for the diagnosis of inflammation, autoimmune disease, anaphylaxis and other illnesss, prevention and treatment.
Vaccination and the mechanism of passive immunity and new generation vaccine
In another embodiment of the disclosure of invention, there is provided for understanding in PCT/US2007/018258 The useful tool of the mechanism and the new high-quality vaccine of exploitation of described vaccination and passive immunity.
In addition, the candidate described in the disclosure of invention can be used for developing binding site vaccine, and it is anti-various The vaccine of pathogen.Whether assessment is excellent for vaccine most important by the binding site characteristic of vaccine-induced antibody.
Prevention and treatment based on therapeutic target
The disclosure of invention also extends into the strategy of the prevention, diagnosis and the new method treated of exploitation associated conditions, can Obtained based on the therapeutic target for identifying, but do not limit and be suitable for the mankind, animal, plant and other biological methods.
As described in PCT/US2007/018258 and the disclosure, (include the mankind or dynamic with organism is present in Thing) in the related candidate (including antibody) of therapeutic target, can be used for infectious disease, autoimmune disease, mistake Diagnosis, prevention (prevention based on therapeutic target) and the treatment of quick disease, cancer, inflammation, obesity and other diseases (is based on and controls The treatment of the property treated target spot).In addition, such candidate is also used as treating infectious disease, autoimmune disease, allergy The drug delivery machinery of disease, cancer, inflammation, obesity and other diseases.
Another strategy be using the similar approach and the disclosure described in PCT/US2007/018258, By the derivant of therapeutic target be used for infectious disease, autoimmune disease, anaphylaxis, cancer, inflammation, obesity and its The diagnosis of its disease, prevention and treatment.
In addition, the derivant of therapeutic target is also used as treating infectious disease, autoimmune disease, allergy The drug delivery machinery of disease, cancer, inflammation, obesity and other diseases.
Another theme of the disclosure of invention is the inactivation granule of the pathogenic agent that pathogenic target spot is had with organism Or fragment or extract are in the infectivity such as the correlation described in PCT/US2007/018258 and in the disclosure Application in the prevention and treatment of disease, autoimmune disease, anaphylaxis, cancer, inflammation, obesity and other diseases.
In addition, the inactivation granule or fragment or extract of pathogen, are also used as treating infectious disease, autoimmune The drug delivery machinery of property disease, anaphylaxis, cancer, inflammation, obesity and other diseases.
Another theme of the disclosure of invention be cause autoimmune disease pathogenic therapeutic target and/or it The application in diagnosis of autoimmune disease, cancer and other associated conditions of correlation candidate thing and/or its derivant.Contain There is the therapeutic target of pathogen or the method acquisition by the disclosure of invention and/or resist the pathogen or therapeutic target The test kit of the antibody of point, can be prepared by various methods well known to those of ordinary skill in the art.Such test kit Can be used for detecting the presence of the antibody in biological sample for antigen.
Disclosure of the invention, can be according to known method (such as by mixing acceptable carrier in pharmacy) Compositionss useful in pharmacy are prepared, said composition contains the therapeutic target and/or its derivant or the therapeutic Any other correlation candidate thing of target spot.The antigen or the other forms of antigen that such composition can be containing effective dose, And/or any other correlation candidate thing of its correlation candidate thing or the therapeutic target, it is acceptable in pharmacy to be formed It is suitable for the compositionss of effective administration.According to the therapeutic target and/or its derivant of the disclosure of invention or described control The using dosage scheme of any other correlation candidate thing of the property treated target spot, can select according to many factors, including described anti- Former positioning and density, the type of patient, species, age, body weight, sex and medical condition；The seriousness of the state of an illness to be treated； Route of administration；The hepatic and renal function of patient；And the particular matter for thus using.Realize material described in the disclosure of invention Concentration is needed available to target site based on used material in the optimal degree of accuracy told on but in the range of no toxicity Dynamic (dynamical) scheme of property.Distribution, balance and exclusion that this includes the material for considering that the disclosure of invention is used.
One embodiment of the disclosure of invention be additionally provided in prevention and treatment new method used in be suitable for office Absorption and the pharmaceutical preparation of parenteral in portion's administration, oral cavity.Said composition contains the therapeutic target and/or its phase Candidate and/or its derivant or any other form of therapeutic target for being accredited as active component are closed, said composition can To be administered in the carrier being administered is traditionally used for extensively changeable therapeutic dosage forms.For example, therapeutic target and/or it Correlation candidate thing and/or its derivant or any other form of therapeutic target, can be applied with peroral dosage form, for example Tablet, capsule (respectively including time controlled released and extended release preparation), pill, powder, granule, elixir (elixirs), tincture (tinctures), solution, suspending agent, syrup and Emulsion, nasal drop, injection, transfusion or the shape being coupled with nano-particle Formula.
Described pharmaceutical composition can be supplied to organism by number of ways, such as subcutaneous administration, closure or not close Local administration, oral administration, intramuscular adminstration, intravenous administration (inject or infuse), Intraperitoneal medication, intramuscular adminstration, Other known to those of ordinary skill in subcutaneous administration, intracavitary administration or percutaneous dosing, inhalation or pharmaceutical field make Use form.
Preparation containing polysaccharide, agglutinin or medical herbs and small molecule
Another theme of the disclosure be for prevention or the infectious disease of therapeutic target host, cancer, The medicine based on binding site of the related biological damage of autoimmune disease, anaphylaxis, inflammation, toxicity or Other diseases Preparation.The pharmaceutical preparation includes：Identification detection candidate or identification target candidate and with their derivant in It is at least one.The pharmaceutical preparation includes：The detection candidate of identification or the target candidate and their derivant of identification are at least One kind, the pharmaceutical preparation is：
1) polysaccharide, including sialic acid and its hydroxyl substituent, the polysaccharide pathogen binding site related to polysaccharide Or the therapeutic target of polysaccharide correlation has same or analogous three dimensional structure：With with the pathogen binding site or described control The property treated target spot competition；2) agglutinin, including but not limited to phytohemagglutinin or medical herbs or antibody, the agglutinin can be with polysaccharide The related therapeutic target of related pathogen binding site or polysaccharide is combined：To block the pathogen binding site or described Therapeutic target；And 3) small molecule, including but not limited to sulfur-containing compound or product：Modify the related pathogen of the polysaccharide The chemical property of the related therapeutic target of binding site or the polysaccharide.
The species of polysaccharide, agglutinin, medical herbs, antibody and small molecule is as described above.Sulfur-containing compound or product include：Sulfur Inorganic compound and organic compound.The inorganic compound of sulfur includes but is not limited to sulfate radical (SO4 2-), sulfate.
The organic compound or product of sulfur is included but is not limited to：Sulfonate, sulphonyl, sulfur (sulfurate), sulfide and Sulfur-containing amino acid.The formula of sulfonate is RSO2O-, wherein R is some organic groups.They are with general formula R SO2The sulfonic acid of OH Conjugate base.Generally, to the compound containing such functional group, ion salt or similar covalent compound, esters, using phase Same title.Sulfur-containing compound or product also include garlic product, including but not limited to Bulbus Allii powder, Oleum Bulbus Allii and Bulbus Allii extract (garlicin (Allicin), Bulbus Allii (Allium sativum), Ajo dilute (Ajoene) etc.).
Sulfonyl is to remove hydroxyl and the organic group that obtains or functional groups by sulfonic acid.Sulfonyl can be write to be had General formula R-S (=O)2- R ', wherein, there are two double bonds between sulfur and oxygen.The title of sulfonyl is typically ended up with-syl, example Such as it is referred to as to tosylate chloride (toluenesulfonyl chloride, CH3CeH4SO2Cl) tosyl chloride (toluene sulphurs Acyl chlorides), or referred to as methylsufonyl chloride (methylsulfonyl chloride, CH3SO2Cl mesyl chloride (first sulphurs) Acyl chlorides).Sulfonyl can be hydrogenated lithium aluminum (LiAlH4) it is reduced into hydrocarbon.
The fabric of anti-infection property pathogen or surface
Another theme of the disclosure of invention is the fabric (fabric) or infection sexually transmitted disease (STD) of anti-infection property pathogen The surface of substance, they can be by the target candidate of the detection candidate of at least one identification or identification be merged in fiber Or the step on fiber and produce；Or following methods and produce：The detection candidate of at least one identification is provided in support material The target candidate of thing or identification, and make granule, then the granule is attached on surface (including but not limited to cloth, facial film (mask), medicated cap or protective spectacles (goggle)).
Embodiment
1. chip carrier
One example of Fig. 1 display chips.A, B, C, D, E and F represent different types of attachment target on the chip Candidate；G represents the example of histochemical stain；And H represents the example of fluorescence staining.
2. the potential therapeutic target of primary screener is carried out using phytohemagglutinin
Fig. 2 and Fig. 3 show the potential biologic treatment carried out as detection candidate using phytohemagglutinin The screening of target spot.The phytohemagglutinin for using be purchased from Vector Laboratories (California, USA), and including：It is special The wheat germ agglutinin (WGA) of opposite sex identification N-ACETYL-D-GLUCOSAMINE, the UEA I (UEA of specific recognition α-Focus I) and specific recognition N- acetyl-D-galactosamine soybean agglutinin (SBA).All of agglutinin with biotin labeling, and With fluorescence (rhodamine (Rhodamine)) the other types of WGA of labelling.The second reagent for the agglutinin of biotin labeling is It is coupled the fluorescence (texas Red (Texas Red)) of streptavidin (streptavidin).
By the tissue slice of the heart of bulb/c adult rats, lung, liver, kidney and spleen respectively with the agglutinin of biotin labeling WGA and SBA cultivates 1 hour (Fig. 2)；By the small bowel tissue sections of bulb/c Neonatal Mouses and adult rats respectively with WGA and UEAI Cultivate (Fig. 3)；And by infect or be uninfected by rhesus rotavirus (RRV) bulb/c neonatal rats liver and small bowel tissue sections (Fig. 4) is cultivated with WGA respectively.After cleaning, streptavidin-texas Red is added, and cultivates 30 minutes, then Cleaning is simultaneously detected with fluorescence microscope.The region of bright dyeing (white) shows that the positive is combined, and the dyeing (grey area) that do not become clear Region be negative combination.
As shown in Fig. 2 the N-ACETYL-D-GLUCOSAMINE represented with the combination of WGA, the heart, lung in bulb/c mices A part of region (rather than all) expression it is very strong；Moderately express in the specific cells of spleen；And do not express in liver and kidney (Fig. 2A).Moderately expressed in the spleen specific cells of bulb/c mices with N- acetyl-D-galactosamine that the combination of SBA is represented, And (Fig. 2 B) is not expressed in heart, lung, liver and kidney.As shown in figure 3, the N- acetyl-D- represented with the combination of WGA and EUA I Glucamine (rather than α-Focus) (Fig. 3 B) expresses very strong in the small intestinal of the 2nd day and the Neonatal Mouse of the 4th day, and into Express weaker in the small intestinal of year Mus.Based on the result of primary screener, N-ACETYL-D-GLUCOSAMINE and N- acetyl-D-galactosamine It is the potential biological marker of relevant disease.Whether they are the binding sites of pathogen, can be by making the pathogen Combine to detect with WGA or SBA with chip, the chip includes people, the health tissues section of animal and plant or as above institute The various kinds of cell system for stating.
The method can be easily extended to other agglutinins to other polysaccharide specificities are attached in said chip, The potential biological marker related to detect other polysaccharide being present in the mankind, animal and other biologies.
3. the identification of the related biological marker of polysaccharide
Fig. 4 shows by making WGA combine to the section of health and diseased tissue the biology mark related to identify polysaccharide The example of note.
Two groups of bulb/c neonatal rats, are administered orally 30 μ l saline (being uninfected by group) and 30 μ l concentration after birth on the 2nd day For 1 × 107The RRV (RRV infected groups) of pfu/ml is processing.The course of disease of the viral infection is 1 week interior, breast after RRV infection Mus suffer from diarrhoea greyish white feces, do not like to take food, and body weight there is no that healthy mice increases it is fast；Neonatal rats of the 30-40% with serious disease There is jaundice.By the 2nd week, all of neonatal rat all occur jaundice, do not like feed, and body weight do not increase；80% suffers from serious disease The neonatal rat of disease is dead.Virus was generally removed in 5 days, can't detect virus at the 5th day.Different number of days after treatment puts to death breast Mus, obtain the tissue sample that blood serum sample, the quick-freezing of intestinal regulating liver-QI and formaldehyde are fixed.
As shown in figure 4, the combination with WGA is represented, always screen in the liver of neonatal rats of the self-infection RRV within mono- week and small intestinal Go out the inflammatory cell or proliferative cell (Fig. 4 B) of the strongly expressed with N-ACETYL-D-GLUCOSAMINE.It is uninfected by the healthy neonatal rat of RRV Liver and the polysaccharide of small intestinal be expressed as negative (Fig. 4 A).Therefore, polysaccharide N-ACETYL-D-GLUCOSAMINE is the latent of inflammation correlation Biological marker.Because many cancers are from the beginning of inflammation, therefore polysaccharide N-ACETYL-D-GLUCOSAMINE can be used as cancer The related potential biological marker of disease.This can by compare WAG be attached on chip health and cancer tissue sections Combination detecting.
Similarly, other biological markers for being related to cancer and Other diseases can have spy by WGA and to other polysaccharide The combination of other agglutinins and chip of the opposite sex, and easily identify, the chip includes human and animal's as above Section or kinds of tumor cells system that health and cancerous tissue or Other diseases are organized.
4. the identification of potential disease inducer
Fig. 5 is shown by anti-rotavirus (RV) polyclonal antibody and agglutinin WGA and the neonatal rat for infecting and being uninfected by RRV Tissue slice combination, identify the example of potential disease inducer.The anti-RV polyclonal antibodies of biotin labeling are purchased from Meridian Life Science, Inc (Mine, USA).In short, by the WGA and biology of rhodamine (Rhodamine) labelling The anti-RV antibody of plain labelling with it is above-mentioned come self-infection and the neonatal rat being uninfected by small bowel tissue sections together with cultivate 1 hour.Cleaning Afterwards, add be coupled Radix Cochleariae officinalises peroxidase (streptavidin-conjugated horseradish peroxidase, HRP streptavidin), cultivates 30 minutes, then cleans, and adds the substrate DAB of HRP and cultivates 15 minutes, so After clean, and detected with routine and fluorescence microscope.The region for being coloured to brown shows that the positive of antibody is combined, and is coloured to The region of shiny red shows that the positive of WGA is combined, and the region (dark areas) of the dyeing that do not become clear is negative combination.
As shown in figure 5, in the small intestinal of the neonatal rat (infection RRV after the 5th day) of infection RRV, anti-RV antibody and expression N- second Goblet cell in the propagation of acyl-D-Glucose amine combines (Fig. 5 B).In acute infection period, virus is generally removed within 1 week, The level of antiviral antibody started to improve from 1 week, reached peak level in 2-3 weeks.Because anti-RV antibodies are in expression N- second Cell in the propagation of acyl-D-Glucose amine, therefore these antibody even can cause inflammation after virus sweep.These are scorching Disease subsequently results in the proliferative reaction of host defense system, further exposes the polysaccharide target spot.Therefore, anti-RV antibody can be with It is struvite inducer, and the autoimmune disease of the tissue or organ of expression N-ACETYL-D-GLUCOSAMINE can be caused. If proliferative inflammation is long lasting for may develop into autoimmune disease.If proliferative inflammation ultimately results in nothing The cell growth of method control, then may develop into cancer.For these reasons, anti-RV antibody can be autoimmune disease With the derivant of cancer.This can pass through the section of the anti-RV antibody of comparison and WGA and the health and diseased tissue for being attached to chip Combination, further to detect.
Similarly, other potential disease inducers can be by other antibody of anti-other pathogen and WGA or to which Its polysaccharide has other agglutinins of specificity and the combination of chip carrier, and easily identifies, the chip carrier is included as above The diseased tissue section of described human and animal, or kinds of tumor cells system.
5. it is used for preventing and treating virus infection and hypersensitive pharmaceutical preparation
Pharmaceutical preparation include it is commercially available synthesis N-acetyl-neuraminate (NeuSAc) (JunKang Biotech Co., Ltd, Guangzhou, China) and Bulbus Allii oil product (Nature ' s Bounty, INC, New York, USA), according to as described below, survey The pharmaceutical preparation is tried to the infection of prevention and treatment virus and hypersensitive effect.
5.1. the prevention of rotavirus infection
Three groups of bulb/c neonatal rats the 1st day after birth, oral administration in the following manner processing, 1) 20 μ l saline (matched group of saline treatment, n=21)；2) Oleum Bulbus Allii of the 20 μ l concentration for 1-5 μ g/g body weight；And 3) 20 μ l contain Neu5Ac The preparation (drug treating group, n=20) of (0.5-2 μ g/g body weight) and Oleum Bulbus Allii (1-5 μ g/g), then as described above, at the 2nd day Infected with RRV.Mice is supported 3 weeks after RRV infection.
Compared with it must compare neonatal rat with salt water pretreatment, with Bulbus Allii oil product or containing Neu5Ac and Bulbus Allii oil product The neonatal rat of preparation pretreatment is not infected.The representational 2 weeks big breasts with saline and the preparation pretreatment are shown in fig. 6 Mus, show the representativeness of the small intestinal of 4 -day-old of neonatal rats in Fig. 6 B (salt water pretreatment) and Fig. 6 C (magistral medicines or medical preconditioning) Histology changes.The result of statistical analysiss is summarized in table 2.
Table 2
**：Virus titer is counted as less than 1: 16 effectively.
The prevention and treatment that 5.2 Avian pneumo-encephalitis virus (NDV) infect
Two groups of SPF chickens (4 weeks ,～2kg) are by collunarium and oral administration 2ml saline (matched group of saline treatment, n=10) Or preparation (drug treating group, n=10) of the 2ml concentration respectively for the Neu5Ac and Oleum Bulbus Allii of 1mg/ml carries out pretreatment, Ran Hou Infected with highly pathogenic NDV within 2nd day.Added once in the 200ml drinking waters of each group at viral metainfective 3rd day respectively 20ml saline or the preparation.The course of disease of the viral disease be in infection 1 week of NDV, chicken diarrhea, do not like to take food, and 50% chicken death.Chicken is supported 8 days after NDV infection.
Infection NDV after the 8th day, 10 to there is 7 death (70%) in the chicken of saline treatment, 2 chickens suffer from abdomen in addition Rush down and do not take food；10 with the preparation process chicken in have 1 death (10%), in addition 2 it is ill.The preparation makes NDV The general mortality rate of infection, is reduced to 10% (odds ratio (Odd Ratio)=0.05,95%CI=0.02-0.96, P=from 70% 0.02).As shown in table 2, the preparation makes the total prevalence rate (dead+ill=invalid) of NDV infection be reduced to from 90% 30%.
The prevention and treatment of the influenza infection of 5.3 Embryo Gallus domesticus
Two groups of Embryo Gallus domesticus are by injecting the Neu5Ac of 100 μ l saline (matched group of saline treatment, n=12) or 100 μ l and big Bulbus Allii oil concentration is the above-mentioned preparation (drug treating group, n=12) of 0.5mg/ml and carries out pretreatment, then connects to allantois within the 2nd day Plant Influenza virus strain H1N1.After virus inoculation, the saline or the system of a 100 μ ls are injected daily in allantois respectively Agent, is collected allantoic fluid after 48 hours in virus inoculation, the virus titer in the liquid is determined by hemagglutination test.
12 virus titers for having 10 with the Embryo Gallus domesticus of saline treatment are as 1: 256, and 12 Embryo Gallus domesticus with saline treatment In have the virus titer of 9 less than 1: 16, in table 2, virus titer is counted as less than 1: 16 effectively.Virus in allantoic fluid Titre be 0 Embryo Gallus domesticus (1 of saline treatment and preparation process 2) not included in statistical analysiss within, it is potential to exclude The failure of virus inoculation.As shown in table 2, the preparation significantly suppress the H1N1 infection of Embryo Gallus domesticus.
The treatment of 5.4 Human Influenzas
Two groups of people experimenters with influenza are by daily 2 times, each collunarium and oral administration 15ml saline (saline treatment Matched group, n=15) or the Neu5Ac and garlic oil concentration of 15ml be the preparation (drug treating group, the n=of 1mg/ml 16) process 2-3 days.The symptom of influenza includes generating heat, has a headache, fatigue, cough, throat pain, rhinorrhea or nasal obstruction, body pain or abdomen Rush down.The sick course of disease is usually 1 week.The flu-like symptom of observation experimenter, observes 7 days daily.With experimenter's phase of saline treatment Relatively, significantly mitigated with the symptom of 15 experimenters of preparation process, and the course of disease shortens (3-4 days being shortened to from 5-7 days). In table 2, these experimenters are counted as effectively.
5.5 the hypersensitive treatment of the mankind
Two groups of human experimenters with seasonal allergic disorder are by each intranasal administration 1-2ml saline daily (at saline The matched group of reason, n=15) or the Neu5Ac and garlic oil concentration of 1-2ml be the preparation (drug treating group, the n of 1mg/ml =15) process 2-3 days.Hypersensitive symptom includes nasal congestion, rhinorrhea, sneeze and rhinocnesmus.The experimenter is observed daily Allergic symptom, observe 3 days.Compared with the experimenter of saline treatment, with the allergy of 14 experimenters of preparation process Symptom significantly mitigates.In table 2, these experimenters are counted as effectively.
Other embodiment than that described above can also be carried out.Therefore, the term and expression is only used for by example To describe the present invention, rather than limit the present invention.It is anticipated that the sheet described in described without departing from here and claim In the case of the spirit and scope of invention, other people can have and above-mentioned different understanding.All patents cited herein, patent go out Version thing and other list of references heres are sentenced its overall mode and are incorporated herein by reference.

Claims (14)

1. it is a kind of identification induced tissue potential disease inducer method, the tissue expression polysaccharide molecule relevant disease target Point or biological marker, the method include：
A) with least one energy and the agglutinin and at least one disease-resistant original antibody and same diseased tissue of glycan molecule specific bond Combine with the health tissues for matching, wherein, the same diseased tissue is the group of the human and animal for infecting the cause of disease Knit, the health tissues that match are the tissue of the human and animal for being uninfected by the cause of disease；
B) as at least one energy and glycan molecule specific bond agglutinin and at least one antibody simultaneously with same disease group Knit and combine, but do not combine with the health tissues for matching, then the antibody is the potential disease inducer for inducing the tissue；
Wherein, the disease is the related biological damage of infectious disease or inflammation.
2. method according to claim 1, wherein, the agglutinin is phytohemagglutinin.
3. method according to claim 2, wherein, the phytohemagglutinin is selected from specific recognition N- acetyl-glucose The Semen sojae atricolor of the wheat germ agglutinin, UEA I of specific recognition fucose and specific recognition N- acetyl-galactosamine of amine Agglutinin.
4. method according to claim 2, wherein, the disease-resistant original antibody is anti-rotavirus antibody.
5. method according to claim 1, wherein, the infectious disease is virus infection.
6. method according to claim 5, wherein, the virus infection is rotavirus, influenza infection and new city Epidemic disease poison infection.
7. the potential disease inducer of the induced tissue identified by the method according to any one in claim 1-6, Wherein, the tissue expression glycan molecule relevant disease target spot or biological marker.
8. potential disease inducer according to claim 7, wherein, the glycan molecule relevant disease target spot or biology It is labeled as：N- acetyl-glucamine and N- acetyl-galactosamine；Or N- acetyl-glucamine and N- acetyl-galactosamine Derivant.
9. potential disease inducer according to claim 8, wherein, the N- acetyl-glucamine in inflammatory cell or Overexpression on proliferative cell；Potential disease inducer of the disease-resistant original antibody for induced tissue, the tissue expression N- Acetyl-glucamine.
10. potential disease inducer according to claim 9, wherein, the inflammation is caused by virus infection；It is described anti- Pathogenic autoantibody is antiviral antibody；The virus is rotavirus, and the antiviral antibody is anti-rotavirus antibody.
The potential disease inducer identified by 11. methods according to any one in claim 1-6 is preparing diagnosis Purposes in the product of the disease related to polysaccharide related target or biological marker；Wherein, the disease is infectious disease Or the biological damage that inflammation is related.
The potential disease inducer identified by 12. methods according to any one in claim 1-6 is preparing treatment With the purposes in the product or drug delivery vehicles of polysaccharide related target or biological marker relevant disease；Wherein, the disease For the related biological damage of infectious disease or inflammation.
13. purposes according to claim 11 or 12, wherein, the polysaccharide related target or biological marker are N- second Acyl-glucamine or N- acetyl-galactosamine.
14. purposes according to claim 11 or 12, wherein, the antibody is anti-rotavirus antibody.
CN200980112434.8A 2008-04-08 2009-04-07 Glycan based array and uses thereof Active CN102016583B (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US4339608P 2008-04-08 2008-04-08
US61/043,396 2008-04-08
PCT/US2009/039810 WO2009126652A2 (en) 2008-04-08 2009-04-07 Glycan based array and uses thereof

Publications (2)

Publication Number Publication Date
CN102016583A CN102016583A (en) 2011-04-13
CN102016583B true CN102016583B (en) 2017-05-10

Family

ID=41162557

Family Applications (1)

Application Number Title Priority Date Filing Date
CN200980112434.8A Active CN102016583B (en) 2008-04-08 2009-04-07 Glycan based array and uses thereof

Country Status (2)

Country Link
CN (1) CN102016583B (en)
WO (1) WO2009126652A2 (en)

Families Citing this family (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102507920A (en) * 2011-09-30 2012-06-20 西安交通大学 Lectin chip and application thereof
CN103131208A (en) * 2013-02-01 2013-06-05 刘中路 Coloring sterilization ointment for anatomic marks in clinical laboratory
CN105051062A (en) * 2013-03-15 2015-11-11 王慧茹 Biological therapeutics for infection-relating disorders or conditions
CN103901212B (en) * 2014-03-28 2015-07-22 西北大学 Glycan microarray for identifying serial liver diseases based on saliva glycan-binding proteins, and application of glycan microarray
WO2016026143A1 (en) 2014-08-22 2016-02-25 Huiru Wang Saccharide-based biomarkers and therapeutics
CN104237448B (en) * 2014-10-09 2016-08-24 成都中医药大学 The thin layer detection method of Radix Anisodi Tangutici root
CN105255868B (en) * 2015-10-23 2018-04-20 广西壮族自治区农业科学院经济作物研究所 Identify bar horse fire fiber crops property method for distinguishing and its molecular labeling, primer pair and kit
FR3044680B1 (en) * 2015-12-02 2017-12-22 Univ Limoges METHOD OF DETECTING CANCER STEM CELLS
CN108003228A (en) * 2018-01-12 2018-05-08 渤海大学 A kind of ginger glycoprotein with ACE inhibitory activity
WO2019208114A1 (en) * 2018-04-25 2019-10-31 パナソニックＩｐマネジメント株式会社 Sensor substrate, method for manufacturing sensor substrate, and detection device
CN109580953B (en) * 2018-11-16 2021-11-05 山西省食品药品检验所(山西省药品包装材料监测中心) Detection device for spearmint colloidal gold doped in mint and mint decoction pieces, preparation method and application
US20230277634A1 (en) * 2020-08-17 2023-09-07 The Regents Of The University Of California Application of microbial glycosidase as an anti-viral therapeutic, prognostic, and diagnostic

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1356552A (en) * 2001-11-08 2002-07-03 郭占军 Bio-chemical protein chip and its preparing process
US20040259142A1 (en) * 2003-06-04 2004-12-23 Imperial College Innovations Limited Products and methods

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1356552A (en) * 2001-11-08 2002-07-03 郭占军 Bio-chemical protein chip and its preparing process
US20040259142A1 (en) * 2003-06-04 2004-12-23 Imperial College Innovations Limited Products and methods

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
凝集素在胃肠道肿瘤及非肿瘤组织病理研究中的应用;黄耀岐 等;《肿瘤》;19840930;第4卷(第5期);216-230页 *
肺肿瘤细胞膜表面糖复合物与组织类型关系的研究;王晓玫 等;《中国肺癌杂志》;20030430;第6卷(第2期);111-115页 *

Also Published As

Publication number Publication date
CN102016583A (en) 2011-04-13
WO2009126652A3 (en) 2009-12-30
WO2009126652A2 (en) 2009-10-15

Similar Documents

Publication Publication Date Title
CN102016583B (en) Glycan based array and uses thereof
US20190388447A1 (en) Glycan-based drugs, therapies and biomarkers
CN110201130A (en) A kind of preparation of Xiaochaihutang extract and mass spectrum fingerprint pattern construction method
US20170232035A1 (en) Polygalacturonan Rhamnogalacturonan1 (PGRG1) Composition
CN102240318A (en) Traditional Chinese medicine for tonifying qi and increasing breast milk
Das et al. Anthelmintic activity of Clerodendrum viscosum
CN107892725A (en) A kind of thizoma curculiginis polysaccharide and its preparation method and application
Du et al. Hepatoprotective and antihyperliposis activities of in vitro cultured Anoectochilus formosanus
CN110898102B (en) Cinnamon extract and traditional Chinese medicine compound for preventing and treating cold injury and application thereof
Ahmad et al. Inhibitory effects of Lepidium sativum polysaccharide extracts on TNF-α production in Escherichia coli-stimulated mouse
CN106706766B (en) A kind of measuring method of the newly-increased detection ingredient of Chinese medicine Tang blade that treating AIDS
Kim et al. The inhibitory effect of a Korean herbal medicine, Zedoariae rhizoma, on growth of cultured human hepatic myofibroblast cells
CN102133302B (en) Verbena extract extracted from verbena and used for curing prostatitis
CN106928376B (en) The separation method of skunk bush polysaccharide and its application
Madhavan et al. PHARMACOLOGICAL AND ANTI CANCER ACTIVITY OF Ipomoea sepiaria METHANOLIC EXTRACT AGAINST PC-3 CELL LINE
CN107320659A (en) It is a kind of to be used to treat Chinese medicine composition of ulcerative colitis and preparation method thereof
CN105358582B (en) A kind of poly- gulonate polysaccharide of Sulfation or its officinal salt and its preparation method and application
Seghatoleslam et al. The potential effects of herbal distillates with hot and cold temperament on cell metabolic activity and growth: a preliminary in vitro study
Vivekanandan et al. Evaluation of the Laxative Activity of Saponin Enriched Hydroethanolic Pericarp Extract of Sapindus emarginatus in Animal Models
SCHRÖDER et al. Cell differentiation process of Artemia sp. larvae tools for natural products testing.
Hu et al. Anti-degranulation response of herbal formula in RBL-2H3 cells
CN103610862A (en) Chinese medicinal herb oral liquid for preventing and treating chicken breathing-type infectious bronchitis
Kuang Recent Advances in Theories and Practice of Chinese Medicine
Dubey et al. Anti-Pyretic Effects Of Aqueous Extract Of Anogeissus Latifolia Roxb In Albino Wistar Rats
CN102266485A (en) Application of rhizoma polygonati aqueous extract in preparation of estrogen drugs

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
TR01 Transfer of patent right
TR01 Transfer of patent right

Effective date of registration: 20240130

Address after: Room 433, No. 42 Guangzhou Road, Zhongxing Street, Development Zone, Nantong City, Jiangsu Province

Patentee after: Jiangsu Mingjian Biopharmaceutical Co.,Ltd.

Country or region after: China

Address before: Illinois, USA

Patentee before: B&H BIOTECHNOLOGIES

Country or region before: U.S.A.