CN109251246A - HIV-1 wide spectrum neutralizing antibody and application thereof - Google Patents
HIV-1 wide spectrum neutralizing antibody and application thereof Download PDFInfo
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- CN109251246A CN109251246A CN201811074636.5A CN201811074636A CN109251246A CN 109251246 A CN109251246 A CN 109251246A CN 201811074636 A CN201811074636 A CN 201811074636A CN 109251246 A CN109251246 A CN 109251246A
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
- C07K16/10—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA viruses
- C07K16/1036—Retroviridae, e.g. leukemia viruses
- C07K16/1045—Lentiviridae, e.g. HIV, FIV, SIV
- C07K16/1063—Lentiviridae, e.g. HIV, FIV, SIV env, e.g. gp41, gp110/120, gp160, V3, PND, CD4 binding site
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
- A61P31/18—Antivirals for RNA viruses for HIV
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
- C07K2317/565—Complementarity determining region [CDR]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Virology (AREA)
- Chemical & Material Sciences (AREA)
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- Medicinal Chemistry (AREA)
- AIDS & HIV (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Oncology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Veterinary Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Communicable Diseases (AREA)
- Public Health (AREA)
- General Chemical & Material Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Hematology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
HIV-1 wide spectrum neutralizing antibody and application thereof.The present invention relates to HIV-1 wide spectrum neutralizing antibodies, include heavy chain variable region and light chain variable region；The antibody specificity combination HIV-1 gp120.The invention further relates to the preparation method of the antibody and purposes.
Description
Technical field
The present invention relates to HIV-1 wide spectrum neutralizing antibody, the antibody specificity combination HIV-1gp120.The invention further relates to
The preparation method and purposes of the antibody.
Background technique
HIV-1 neutralizing antibody can effectively prevent HIV-1 from tying the CD4+T cell for being combined into people.Therefore, as right
A kind for the treatment of means of HIV-1 the infected, AIDS research field are being dedicated to developing always in potent HIV-1 wide spectrum and are resisting
Body.
Monoclonal antibody b12 obtained by phage display library technology earliest, but in being only capable of and about 40%
Know HIV-1 virus[1].Since 2010, have benefited from the progress of the technologies such as single B cell sorting and deep sequencing,
A variety of human monoclonal antibodies with anti-HIV-1 wide spectrum neutralization activity are isolated from HIV-1 the infected, are such as directed to CD4
The VRC01 of binding site[2], for the PG9/PG16 and PGT121 of variable region V1/V2 and V3[3,4], for membrane-proximal region (MPER,
Membrane proximal external region) 10E8 antibody[5]With the 35O22 for being directed to gp120-gp41 intersection[6]Deng.
In view of HIV-1, there are a variety of clade and worldwide there are different Major Epidemic strains, groups for different regions
Closing using two or more wide spectrum neutralizing antibodies is clearly a kind of therapeutic strategy for having more potentiality.Therefore, AIDS research is led
There is still a need for develop new strong effect wide-spectrum reactivity HIV-1 neutralizing antibody in domain.
Summary of the invention
Present invention aim to address HIV-1 strain degree of variation is big, traditional monoclonal antibody neutralization width is limited
Problem provides one plant of HIV-1 wide spectrum neutralizing antibody and application thereof.
Technical solution of the present invention
Present invention firstly provides a kind of isolated human monoclonal antibodies or functional antibody fragment, wherein antibody packet
Containing heavy chain variable region and light chain variable region:
Wherein the heavy chain variable region includes and is respectively corresponding to contained in amino acid sequence shown in SEQ ID NO:2
VHCDR1, VHCDR2 and VHCDR3 of VHCDR1, VHCDR2 and VHCDR3, and
Wherein the light chain variable region includes and is respectively corresponding to contained in amino acid sequence shown in SEQ ID NO:4
VLCDR1, VLCDR2 and VLCDR3 of VLCDR1, VLCDR2 and VLCDR3, and
Wherein the antibody is the neutralizing antibody for specifically binding HIV-1gp120.The antibody can block HIV-1 to tie
It is combined into target cell.
The heavy chain variable region includes amino acid 26-33,51-57 and the 96-113 for being respectively corresponding to SEQ ID NO:2
VHCDR1, VHCDR2 and VHCDR3.
And the heavy chain variable region includes amino acid sequence shown in SEQ ID NO:2 or has extremely with SEQ ID NO:2
The amino acid sequence of few 85% phase same sex.
The light chain variable region includes amino acid 27-37,55-57 and the 94-102 for being respectively corresponding to SEQ ID NO:4
VLCDR1, VLCDR2 and VLCDR3.
And the light chain variable region includes amino acid sequence shown in SEQ ID NO:4 or has extremely with SEQ ID NO:4
The amino acid sequence of few 85% phase same sex.
Present invention colleague provides a kind of antibody fragment, selected from Fab segment, Fab ' segment, ' 2 F (ab) segment, single-stranded
Fv albumen (scFv) and the stable Fv albumen (dsFv) of disulfide bond.
It the present invention also provides the nucleic acid molecules of encoding such antibodies or antibody fragment and include at least one above-mentioned core
The expression vector of acid molecule.
The present invention also provides the host cells through at least one above-mentioned nucleic acid molecules or expression vector conversion.
The present invention also provides use at least one above-mentioned nucleic acid molecules or expression vector or host cell to produce antibody
Method, this method comprises:
(i) host cell is converted with nucleic acid molecules as claimed in claim 7 or expression vector according to any one of claims 8,
(ii) in the case where being suitble to the nucleic acid molecules or expression vector to express incubation step (i) described conversion host
Cell, and
(iii) it separates and is purified by the nucleic acid molecules or expression vector institute table from the host cell that step (ii) is cultivated
The antibody or antibody fragment reached.
The present invention also provides the pharmaceutical compositions comprising at least one above-mentioned antibody or antibody fragment.
In some embodiments, the heavy chain variable region of antibody of the invention includes to be respectively corresponding to SEQ ID NO:2's
VHCDR1, VHCDR2 and VHCDR3 of amino acid 26-33,51-57 and 96-113, and the light chain variable region packet of antibody of the invention
VLCDR1, VLCDR2 and VLCDR3 containing amino acid 27-37,55-57 and the 94-102 for being respectively corresponding to SEQ ID NO:4.This
The antibody of invention is the wide spectrum neutralizing antibody for specifically binding HIV-1gp120.
The present invention also provides the methods of the HIV-1 infection of detection human subjects, comprising:
(i), the biological sample from the object is made to be in contact with antibody of the present invention or antibody fragment；
(ii), it determines and whether there is the immune complex formed by the antibody or the antibody fragment in the sample,
Wherein there is the immune complex and shows that the object has HIV-1 infection.
In some embodiments, the sample is immobilized onto solid substance before carrying out the contact.Another
In a little embodiments, the antibody or antibody fragment are immobilized onto solid substance before carrying out the contact.In some realities
It applies in mode, fluorescent marker, enzyme label or radioactive label is marked in the antibody or antibody fragment.In some embodiments
In, immune complex is detected using the secondary antibody for specifically binding the antibody or antibody fragment.In other embodiments
In, immune complex is detected using the secondary antibody of the antigen of specific binding HIV-1.The present invention also provides for detecting the mankind
The kit of the HIV-1 infection of object, it includes antibody of the invention or antibody fragments.
The present invention further provides the methods prevented or the HIV-1 for the treatment of human subjects infects, including apply to the object
With a effective amount of at least one antibody or antibody fragment of the invention or pharmaceutical composition of the invention.In some embodiments
In, the object suffers from acquired immunodeficiency syndrome (AIDS).It in some embodiments, further include being applied to the object
With the antiviral drugs of at least one anti-HIV-1.
It is infected in preparation for detecting the HIV-1 of human subjects the present invention also relates to antibody of the invention or antibody fragment
Purposes in kit or the pharmaceutical composition of the HIV-1 infection for preventing or treating human subjects.
The advantages of the present invention: the present invention provides one plant of HIV-1 wide spectrum neutralizing antibodies, can neutralize extensively
The HIV-1 virus of each hypotype/reform patterns, neutralization width have reached 57%.
Detailed description of the invention
Fig. 1 show the comparison analysis result of F6 heavy chain of antibody and light-chain amino acid sequence and corresponding family gene.
Fig. 2 is antibody F6 and HIV-1gp120 and gp140 combination schematic diagram.
Sequence explanation
SEQ ID NO:1 is the coded sequence of antibody F6 heavy chain variable region.
SEQ ID NO:2 is the amino acid sequence of antibody F6 heavy chain variable region.
SEQ ID NO:3 is the coded sequence of antibody F6 light chain variable region.
SEQ ID NO:4 is the amino acid sequence of antibody F6 light chain variable region.
Preservation information
Carry comprising antibody F6 heavy chain gene expression vector escherichia coli (Escherichia coli) in
In September, 2018 is preserved in CGMCC, and deposit number is CGMCC No.16431.
Carry comprising antibody F6 light chain gene expression vector escherichia coli (Escherichia coli) in
In September, 2018 is preserved in CGMCC, and deposit number is CGMCC No.16432.
Specific embodiment
The present inventor's isolated one plant of monoclonal antibody out of HIV-1 Chinese epidemic strain the infected's body, concurrently
Existing its has wide spectrum neutralization activity to a variety of HIV-1, which is named as F6.
On the one hand, the present invention provides isolated human monoclonal antibodies, it includes heavy chain variable region and light chain variable region,
Described in heavy chain variable region include be respectively corresponding to VHCDR1, VHCDR2 contained in amino acid sequence shown in SEQ ID NO:2
With VHCDR1, VHCDR2 and VHCDR3 of VHCDR3, and
Wherein the light chain variable region includes and is respectively corresponding to contained in amino acid sequence shown in SEQ ID NO:4
VLCDR1, VLCDR2 and VLCDR3 of VLCDR1, VLCDR2 and VLCDR3, and
Wherein the antibody is the neutralizing antibody for specifically binding HIV-1gp120.
In some embodiments, the heavy chain variable region includes the amino acid 26- for being respectively corresponding to SEQ ID NO:2
33, VHCDR1, VHCDR2 and VHCDR3 of 51-57 and 96-113, and
In some embodiments, the light chain variable region includes the amino acid 27- for being respectively corresponding to SEQ ID NO:4
37, VLCDR1, VLCDR2 and VLCDR3 of 55-57 and 94-102.
Herein, when being related to antibody of the invention, " heavy chain variable region includes to correspond to SEQ ID NO:2 institute
Show this statement of the VHCDR1 " of VHCDR1 contained in amino acid sequence refer to the VHCDR1 in the antibody heavy chain variable region with
VHCDR1 amino acid sequence having the same contained in amino acid sequence shown in the SEQ ID NO:2.For example, according to IMGT
Database analysis, the 26-33 amino acids of the VHCDR1 of antibody F6 as the heavy chain variable region as shown in SEQ ID NO:2
(GVSLRGYY) it forms, then above-mentioned statement refers to the VHCDR1 of the heavy chain variable region of antibody of the invention also by GVSLRGYY group
At.
Herein, when being related to antibody of the invention, " light chain variable region includes to correspond to SEQ ID NO:4 institute
Show this statement of the VLCDR1 " of VLCDR1 contained in amino acid sequence refer to the VLCDR1 in the antibody's light chain variable region with
VLCDR1 amino acid sequence having the same contained in amino acid sequence shown in the SEQ ID NO:4.For example, according to IMGT
Database analysis, the 27-37 amino acids of the VLCDR1 of antibody F6 as the light chain variable region as shown in SEQ ID NO:4
(QSLLNRNGDNY) form, then above-mentioned statement refer to the VLCDR1 of the light chain variable region of antibody of the invention by
QSLLNRNGDNY composition.
Herein, VHCDR, HCDR and CDRH have same meaning, refer to that the complementation of antibody heavy chain variable region is determined
Determine cluster, is used interchangeably.VLCDR, LCDR and CDRL have same meaning, refer to that the complementary of antibody's light chain variable region determines
Cluster is used interchangeably.
In some embodiments, the heavy chain variable region includes amino acid sequence or and SEQ shown in SEQ ID NO:2
ID NO:2 has the amino acid sequence or described heavy of at least 85%, at least 90%, at least 95% or the higher order column phase same sex
Chain variable region amino acid sequence as shown in SEQ ID NO:2 or with SEQ ID NO:2 have at least 85%, at least 90%, at least
95% or the higher order column phase same sex amino acid sequence composition.In some embodiments, the heavy chain variable region includes and SEQ
ID NO:2 have about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or
The amino acid sequence of about 99% sequence identity.In some preferred embodiments, the heavy chain variable region includes institute in table 1
VHCDR1, VHCDR2 and the VHCDR3 shown.
In some embodiments, the light chain variable region includes amino acid sequence or and SEQ shown in SEQ ID NO:4
ID NO:4 has the amino acid sequence of at least 85%, at least 90%, at least 95% or the higher order column phase same sex.In some implementations
In mode, the light chain variable region include with SEQ ID NO:4 have about 90%, about 91%, about 92%, about 93%, about 94%,
The amino acid sequence of about 95%, about 96%, about 97%, about 98% or about 99% sequence identity.In some preferred embodiment party
In formula, the light chain variable region includes VLCDR1, VLCDR2 and VLCDR3 shown in table 1.
In some embodiments, antibody of the invention is IgG.In other embodiments, antibody of the invention is
IgM.In some other embodiment, antibody of the invention is IgA.
The present invention further provides isolated antibody fragments, are the functional fragment of aforementioned antibody of the invention, energy
Enough specifically bind HIV-1gp120.In some embodiments, antibody fragment of the invention be selected from Fab segment, Fab ' segment,
' 2 F (ab) segment, single chain Fv protein (scFv) and the stable Fv albumen (dsFv) of disulfide bond.Preferably, antibody piece of the invention
Section has wide spectrum neutralization activity to a variety of HIV-1.
The present invention also provides isolated polypeptide, the polypeptide is immunoglobulin heavy chain variable area, can be used for constructing spy
The opposite sex combines HIV-1gp120 and has the antibody of wide spectrum neutralization activity to HIV-1.In some embodiments, the polypeptide packet
VHCDR1, VHCDR2 and VHCDR3 containing amino acid 26-33,51-57 and the 96-113 for being respectively corresponding to SEQ ID NO:2.?
In other embodiments, the polypeptide include with SEQ ID NO:2 have about 90%, about 91%, about 92%, about 93%, about
94%, the amino acid sequence of about 95%, about 96%, about 97%, about 98% or about 99% sequence identity.In some embodiment party
In formula, the VLCDR1 of the polypeptide amino acid 27-37,55-57 and 94-102 comprising being respectively corresponding to SEQ ID NO:4,
VLCDR2 and VLCDR3.In other embodiments, the polypeptide include with SEQ ID NO:4 have about 90%, about 91%,
The amino acid sequence of about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% sequence identity
Column.
On the other hand, the present invention provides isolated nucleic acid molecules, encodes aforementioned antibody or antibody fragment of the invention
Or polypeptide.In some embodiments, nucleic acid molecules of the invention are operably coupled to promoter.
The present invention also provides expression vectors, and it includes at least one aforementioned nucleic acid molecules of the invention.
The present invention also provides isolated host cells, by least one aforementioned nucleic acid molecules or expression vector of the invention
Conversion.
On the other hand, the present invention provides the method for production antibody, comprising:
(i) host cell is converted with aforementioned at least one nucleic acid molecules of the invention or expression vector,
(ii) host cell of the conversion is cultivated in the case where being suitble to the nucleic acid molecules or expression vector is expressed, and
(iii) it separates and purifies the antibody as expressed by the nucleic acid molecules or expression vector or antibody fragment.
The invention further relates to the isolated antibody or antibody fragment that the method by aforementioned present invention obtains, can be special
The opposite sex combines HIV-1 gp120.Preferably, the isolated antibody or antibody fragment pair obtained by the method for aforementioned present invention
A variety of HIV-1 have wide spectrum neutralization activity.
On the other hand, the present invention provides pharmaceutical composition, and it includes at least one aforementioned antibody or antibody of the invention
Segment and pharmaceutical acceptable carrier.
On the other hand, the present invention provides the method for the HIV-1 infection of detection human subjects, comprising:
(i) biological sample from the object is made to be in contact with aforementioned antibody or antibody fragment of the invention, and
(ii) it determines and whether there is the immune complex formed by the antibody or the antibody fragment in the sample,
Wherein there is the immune complex and shows that the object has HIV-1 infection.
In some embodiments for the method that the HIV-1 of detection human subjects of the invention infects, in step (i),
The sample is immobilized onto solid substance, and the contact include the liquid containing the antibody or antibody fragment is added to it is described
Solid substance.In some embodiments, fluorescent marker, enzyme label or radioactivity mark is marked in the antibody or antibody fragment
Note.In other embodiments, in step (ii), make the solid substance and specifically bind the antibody or antibody piece
First binding partners of section are in contact.In some embodiments, first binding partners are described in specific binding
The secondary antibody of antibody or antibody fragment.
In other embodiments for the method that the HIV-1 of detection human subjects of the invention infects, in step (i)
In, the antibody or antibody fragment are immobilized onto solid substance, and the contact includes adding to the liquid containing the sample
The solid substance.In some embodiments, in step (ii), make the solid substance and specifically bind HIV-1's
Second binding partners of antigen are in contact.In some embodiments, second binding partners are specific bindings
The secondary antibody of the antigen of HIV-1.In some embodiments, the secondary antibody specifically binds HIV-1gp120.One
In a little embodiments, the antibody or the antibody fragment HIV-1 in conjunction with the secondary antibody of the specific binding HIV-1 antigen
Different epitopes on antigen.
In some embodiments for the method that the HIV-1 of detection human subjects of the invention infects, the object is come from
Biological sample be whole blood, blood plasma, serum, haemocyte or haemocyte lysate.
In some embodiments for the method that the HIV-1 of detection human subjects of the invention infects, the object is come from
Biological sample contain haemocyte, and wherein the method further includes before step (i), in or after the process, will
The biological sample is in contact with the third binding partners for specifically binding the haemocyte.In some embodiments,
The third binding partners are the antibody for specifically binding the haemocyte.In some specific embodiments, the blood
Cell is lymphocyte, such as T cell, such as CD4+T cell.In other specific embodiments, the haemocyte is
Monocyte.In some specific embodiments, the third binding partners are on the specific binding haemocyte
The antibody of characteristic markers.
On the other hand, the present invention relates to aforementioned antibody or antibody fragment of the invention to prepare for detecting human subjects
HIV-1 infection kit in purposes.
On the other hand, the present invention also provides the kit of the HIV-1 infection for detecting human subjects, it includes aforementioned
Antibody or antibody fragment of the invention.
On the other hand, the present invention also provides the methods prevented or the HIV-1 for the treatment of human subjects infects, including to described
Object applies a effective amount of at least one aforementioned antibody or antibody fragment of the invention or pharmaceutical composition of the invention.Some
In embodiment, the object suffers from acquired immunodeficiency syndrome (AIDS).In some embodiments, side of the invention
Method further comprises that the antiviral drugs of at least one anti-HIV-1 is applied to the object.
On the other hand, the invention further relates to aforementioned antibody or antibody fragment of the invention to prepare for preventing or treating
Purposes in the pharmaceutical composition of the HIV-1 infection of human subjects.In some embodiments, the object is exempted from acquired
Epidemic disease deficit syndrome (AIDS).
Following embodiment has no intention to limit the scope of the invention in any way for illustrating the present invention.
Embodiment
Embodiment: wide spectrum neutralizing antibody F6
The isolated antibody F6 out of HIV-1 infection in Chinese body
Inventor's isolated one plant of monoclonal antibody out of only one China HIV-1 the infected's body, is named as F6.It carries
The escherichia coli of the expression vector (F6K) of the expression vector (F6H) and light chain gene of heavy chain gene comprising antibody F6
(Escherichia coli) is preserved in respectively with deposit number CGMCC No.16431 and CGMCC No.16432 Chinese common micro-
Biological inoculum preservation administrative center (China General Microbiological Culture Collection Center,
CGMCC) (Yard 1, BeiChen xi Road, Chaoyang District, Beijing City).The coded sequence of contained heavy chain variable region is SEQ ID NO:1 in F6H,
The amino acid sequence of its heavy chain variable region encoded is SEQ ID NO:2.The coded sequence of contained light chain variable region is in F6K
SEQ ID NO:3, the amino acid sequence of the light chain variable region of coding are SEQ ID NO:4.Table 1 give antibody F6 heavy chain and
The structure and CDR information of light chain variable region.
The sequence of F6 antibody is analyzed
As shown in Figure 1, inventor using antibody gene analytical database IMGT V-QEST server (http: //
Www.imgt.org/IMGT_vquest/vquest? livret=0&Option=humanIg F6 antibody gene) is analyzed.
F6 heavy chain of antibody belongs to IgHV4-34*01 family, and CDRH3 is 18 amino acid.Light chain belongs to IgKV2-28*01 family,
CDRL3 is 9 amino acid.
The preparation of F6 antibody
The escherichia coli of the escherichia coli and light chain expression vector F6K of heavy chain expression vector F6H will be carried, point
It is not inoculated in 100ml to contain in the LB culture medium (Amersham Products) of 50 μ g/ml kanamycins, 37 DEG C, 200rpm vibration
Swing culture 16 hours.Expression vector plasmid is extracted using the Plasmid Midi Kit kit of Omega company.Turned using PEI
Transfection reagent (Polysciences Products) is with the heavy chain of equivalent and light chain expression vector cotransfection 293F cell, 8%CO2,
37 DEG C are cultivated 6 days.Antibody F6 is obtained using Protein A affinity column (GE health Products) purifying.It utilizes
NanoDrop2000 ultramicrospectrophotometer (Thermo Products) measures antibody concentration, and 4 DEG C of placements are to be detected.
The detection of F6 antibody binding capacity
The binding ability of antibody is determined by ELISA method.Antigen protein CN54gp120 and CN54gp140 are used
PBS is diluted to 2 μ g/ml, every 100 μ l of hole and is coated in 96 hole elisa plates (Corning Costar Products), 4 DEG C of mistakes
Night.With PBS-T solution (0.05% Tween-20) board-washing 5 times, 250 μ l confining liquids (PBS, 2% BSA+5%milk) is added in every hole
Room temperature is closed 1 hour.PBS-T board-washing 3 times, by monoclonal antibody using 10 μ g/ml as initial concentration, carrying out 5 times with confining liquid is
Column dilution.100 μ l samples are taken to be added in elisa plate respectively, 37 DEG C are incubated for 1 hour.PBS-T board-washing 5 times, every hole is added 100
Goat anti-Human IgG (H+L) (Beijing Zhong Shan Golden Bridge biology of horseradish peroxidase-labeled of the μ l after confining liquid 1:5000 dilution
Technology Co., Ltd.'s product), 37 DEG C are incubated for 1 hour.PBS-T board-washing 5 times, 100 μ l TMB chromogenic substrates (Beijing gold person of outstanding talent is added
Pharmacy stock Co., Ltd's product), room temperature is protected from light colour developing 20 minutes.Every hole is directly added into (the bold and unconstrained pharmacy of Beijing gold of 50 μ l terminate liquids
Limited liability company's product) reaction is terminated, microplate reader reads the absorbance value of 450nm wavelength.As a result such as Fig. 2 is shown, antibody F6
It can be in conjunction with gp120 and gp140 antigen protein.
The detection of F6 antibody neutralising capacity
By in TZM-bl/ pseudovirus and testing[7]Determine the neutralising capacity of antibody.With DMEM growth medium
(Hyclone Products) dilute monoclonal antibody gradient series, and the antibody and 50 μ l that 100 μ l have diluted contain 200
TCID50Pseudovirus be added 96 orifice plates in, 5%CO2, 37 DEG C are incubated for 1 hour.1 × 10 will be contained4A TZM-bl cell and 11 μ
The cell liquid of g/ml DEAE-dextran (Sigma Products) is added in 96 orifice plates, at the same be arranged cell controls (containing only
TZM-bl cell) and virus control (containing only TZM-bl cell and pseudovirus), 5%CO237 DEG C are cultivated 48 hours.It utilizes
Bright-Glo luciferase reagent kit (Promega Products) detects luciferase reaction, calculates 50%
Inhibit dosage.As shown in table 2, F6 antibody can neutralize the HIV-1 virus of different subtype, be wide spectrum neutralizing antibody.
In table .2 antibody F6 and the ability of HIV-1 pseudovirus
F6 antibody neutralization effect (geometric mean): 12.15 μ g/ml, neutralization effect by antibody half-inhibitory concentration
(IC50) it indicates.
F6 antibody neutralization width (< 50 μ g/ml): 57%, viral percentage of the neutralization width by neutralization effect less than 50 μ g/ml
Number indicates.
Protection scope of the present invention should include obviously replacement and combination for those skilled in the art.
Descriptions above is merely to illustrate the present invention, it is clear that without departing from the spirit and substance in the present invention, people from this field
Member can make a variety of modifications and changes to the present invention, thus these modifications and changes equally this application claims range
It is interior.
Bibliography
1.Dennis R.Burton,Jayashree Pyati,Raju Koduri,Stephen J.Sharp,George
B. Thornton,Paul W.H.1.Parren,Lynette S.W.Sawyer,R.Michael Hendry,Nancy
Dunlop, Peter L.Nara,Michael Lamacchia,Eileen Garratty,E.Richard Stiehm,
Yvonne J.Bryson, Yunzhen Cao,John P.Moore,David D.Ho,Carlos F.Barbas
III.Efficient neutralization of primary isolates of HIV-1by a recombinant
human monoclonal antibody.Science.1994. 266:1024-1027.
2.Xueling Wu,Zhi-Yong Yang,Yuxing Li,Carl-Magnus Hogerkorp,William
R.Schief, Michael S.Seaman,Tongqing Zhou,Stephen D.Schmidt,Lan Wu,Ling Xu,
Nancy S.Longo, Krisha McKee,Sijy O’Dell,Mark K.Louder,Diane L.Wycuff,Yu Feng,
Martha Nason, Nicole Doria-Rose,Mark Connors,Peter D.Kwong,Mario Roederer,
Richard T.Wyatt,Gary J.Nabel,John R.Mascola.Rational design of envelope
identifies broadly neutralizing human monoclonal antibodies to HIV-
1.Science.2010.329:856-861.
3.Jason S.McLellan,Marie Pancera1,Chris Carrico,Jason Gorman,Jean-
Philippe Julien,Reza Khayat,Robert Louder,Robert Pejchal,Mallika Sastry,
Kaifan Dai,Sijy O’Dell1, Nikita Patel,Syed Shahzad-ul-Hussan,Yongping Yang,
Baoshan Zhang,Tongqing Zhou, Jiang Zhu,Jeffrey C.Boyington,Gwo-Yu Chuang,
Devan Diwanji,Ivelin Georgiev,Young Do Kwon,Doyung Lee,Mark K.Louder,
Stephanie Moquin,Stephen D.Schmidt,Zhi-Yong Yang,Mattia Bonsignori,John
A.Crump,Saidi H.Kapiga,Noel E.Sam,Barton F.Haynes, Dennis R.Burton,Wayne
C.Koff,Laura M.Walker,Sanjay Phogat,Richard Wyatt,Jared Orwenyo,Lai-Xi Wang,
James Arthos,Carole A.Bewley,John R.Mascola,Gary J.Nabel, William R.Schief,
Andrew B.Ward,Ian A.Wilson,and Peter D.Kwong.Structure of HIV-1 gp120V1/V2
domain with broadly neutralizing antibody PG9.Nature.2011.480:336-343.
4.Jean-Philippe Julien,Devin Sok,Reza Khayat,Jeong Hyun Lee,Katie
J.Doores, Laura M.Walker,Alejandra Ramos,Devan C.Diwanji,Robert Pejchal,
Albert Cupo,Umesh Katpally,Rafael S.Depetris,Robyn L.Stanfield,Ryan McBride,
Andre J.Marozsan,James C. Paulson,Rogier W.Sanders,John P.Moore,Dennis
R.Burton,Pascal Poignard,Andrew B. Ward,Ian A.Wilson.Broadly Neutralizing
Antibody PGT121 Allosterically Modulates CD4 Binding via Recognition of the
HIV-1 gp120 V3 Base and Multiple Surrounding Glycans. PLoS Pathog.2013.9:
e1003342.
5.Jinghe Huang,Gilad Ofek,Leo Laub,Mark K.Louder,Nicole A.Doria-Rose,
Nancy S.Longo,Hiromi Imamichi,Robert T.Bailer,Bimal Chakrabarti,Shailendra
K.Sharma,S. Munir Alam,TaoWang,Yongping Yang,Baoshan Zhang,Stephen
A.Migueles,Richard Wyatt, Barton F.Haynes,Peter D.Kwong,John R.Mascola,Mark
Connors.Broad and potent neutralization of HIV-1 by a gp41-specific human
antibody.Nature.2012.491:406-412.
6.Jinghe Huang,Byong H.Kang,Marie Pancera,Jeong Hyun Lee,Tommy Tong,
Yu Feng,Ivelin S.Georgiev,Gwo-Yu Chuang,Aliaksandr Druz,Nicole A.Doria-Rose,
Leo Laub, Kwinten Sliepen,Marit J.van Gils,Alba Torrents de laRonald
Derking,Per-Johan Klasse,Stephen A.Migueles,Robert T.Bailer,Munir Alam,Pavel
Pugach,Barton F.Haynes, Richard T.Wyatt,Rogier W.Sanders,James M.Binley,
Andrew B.Ward,John R.Mascola, Peter D.Kwong,Mark Connors.Broad and potent
HIV-1 neutralization by a human antibody that binds the gp41-120
interface.Nature.2014.515:138-142.
7.Ming Li,Feng Gao,John R.Mascola,Leonidas Stamatatos,Victoria
R.Polonis, Marguerite Koutsoukos,Gerald Voss,Paul Goepfert,Peter Gilbert,
Kelli M.Greene, Miroslawa Bilska,Denise L.Kothe,Jesus F.Salazar-Gonzalez,
Xiping Wei,Julie M.Decker, Beatrice H.Hahn,David C.Montefiori.Human
Immunodeficiency Virus Type 1 env Clones from Acute and Early Subtype B
Infections for Standardized Assessments of Vaccine-Elicited Neutralizing
Antibodies.J Virol.2005.79:10108-10125。
Sequence table
<110>Nankai University
<120>HIV-1 wide spectrum neutralizing antibody and application thereof
<141> 2018-09-14
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 372
<212> DNA
<213>Genus Homo species home sapiens (Homo sapiens)
<400> 1
caggtgcagc tacagcagtg gggcacagga ctgttgaagc cctcggagac cctgtccctc 60
acgtgcgctg tctatggtgt gtcgttgaga ggctattact ggacgtggat ccgccagtcc 120
ccaaaaaagg gcctggagtg gattggggaa attgatgaga ttggaaggac gaaatacagt 180
cagtccctca ggagtcgggc caccctctca atagacacgt ccaagaaaca attctccctg 240
aggctgacgt ccgtgaccgc cgccgacatg gcgacctatt attgtgcgag atggcgtcta 300
atgatggtgg acgaagtgac aaggcacggg atggacgtct ggagtcaggg gaccatggtc 360
accgtctcct ca 372
<210> 2
<211> 124
<212> PRT
<213>Genus Homo species home sapiens (Homo sapiens)
<400> 2
Gln Val Gln Leu Gln Gln Trp Gly Thr Gly Leu Leu Lys Pro Ser Glu
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Val Tyr Gly Val Ser Leu Arg Gly Tyr
20 25 30
Tyr Trp Thr Trp Ile Arg Gln Ser Pro Lys Lys Gly Leu Glu Trp Ile
35 40 45
Gly Glu Ile Asp Glu Ile Gly Arg Thr Lys Tyr Ser Gln Ser Leu Arg
50 55 60
Ser Arg Ala Thr Leu Ser Ile Asp Thr Ser Lys Lys Gln Phe Ser Leu
65 70 75 80
Arg Leu Thr Ser Val Thr Ala Ala Asp Met Ala Thr Tyr Tyr Cys Ala
85 90 95
Arg Trp Arg Leu Met Met Val Asp Glu Val Thr Arg His Gly Met Asp
100 105 110
Val Trp Ser Gln Gly Thr Met Val Thr Val Ser Ser
115 120
<210> 3
<211> 336
<212> DNA
<213>Genus Homo species home sapiens (Homo sapiens)
<400> 3
gatattgtga tgactcagtc tcctttgtcc ctgtccgtcg cccctggaga ggcggcctcc 60
atctcctgca ggtctacaca gagcctcctc aataggaacg gagacaatta tttggagtgg 120
tacttacgga ggccagggcg gtctccacaa ctcctcatct atttgggttc tgaacgggcc 180
ttgggggtcc ctgacaggtt cagtggcagt gggtcaggca gagattttac actgaaaatc 240
agcagagtgg aggctcagga tgtagggacc tattactgct tgcaaactcg acaaggtgcg 300
ttcacttttg gccaggggac caagctggag atcaaa 336
<210> 4
<211> 112
<212> PRT
<213>Genus Homo species home sapiens (Homo sapiens)
<400> 4
Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Ser Val Ala Pro Gly
1 5 10 15
Glu Ala Ala Ser Ile Ser Cys Arg Ser Thr Gln Ser Leu Leu Asn Arg
20 25 30
Asn Gly Asp Asn Tyr Leu Glu Trp Tyr Leu Arg Arg Pro Gly Arg Ser
35 40 45
Pro Gln Leu Leu Ile Tyr Leu Gly Ser Glu Arg Ala Leu Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Arg Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Gln Asp Val Gly Thr Tyr Tyr Cys Leu Gln Thr
85 90 95
Arg Gln Gly Ala Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
100 105 110
Claims (15)
1. isolated human monoclonal antibodies or functional antibody fragment, wherein antibody includes heavy chain variable region and light chain variable
Area:
Wherein the heavy chain variable region include be respectively corresponding to VHCDR1 contained in amino acid sequence shown in SEQ ID NO:2,
VHCDR1, VHCDR2 and VHCDR3 of VHCDR2 and VHCDR3, and
Wherein the light chain variable region include be respectively corresponding to VLCDR1 contained in amino acid sequence shown in SEQ ID NO:4,
VLCDR1, VLCDR2 and VLCDR3 of VLCDR2 and VLCDR3, and
Wherein the antibody is the neutralizing antibody for specifically binding HIV-1gp120.
2. isolated human monoclonal antibodies according to claim 1 or functional antibody fragment, wherein the heavy chain
Variable region include be respectively corresponding to SEQ ID NO:2 amino acid 26-33,51-57 and 96-113 VHCDR1, VHCDR2 and
VHCDR3。
3. isolated human monoclonal antibodies according to claim 1 or 2 or functional antibody fragment, wherein described heavy
Chain variable region includes amino acid sequence shown in SEQ ID NO:2 or has the amino of at least 85% phase same sex with SEQ ID NO:2
Acid sequence.
4. isolated human monoclonal antibodies according to claim 1 or functional antibody fragment, wherein the light chain
Variable region include be respectively corresponding to SEQ ID NO:4 amino acid 27-37,55-57 and 94-102 VLCDR1, VLCDR2 and
VLCDR3。
5. isolated human monoclonal antibodies according to claim 4 or functional antibody fragment, wherein the light chain
Variable region includes amino acid sequence shown in SEQ ID NO:4 or has the amino acid of at least 85% phase same sex with SEQ ID NO:4
Sequence.
6. the antibody fragment of any one of claim 1-5 is selected from Fab segment, Fab ' segment, ' 2 F (ab) segment, scFv
Albumen (scFv) and the stable Fv albumen (dsFv) of disulfide bond.
7. isolated nucleic acid molecules encode antibody or antibody fragment of any of claims 1-6.
8. expression vector, the nucleic acid molecules as claimed in claim 7 comprising being operably coupled to promoter.
9. isolated host cell is converted by nucleic acid molecules as claimed in claim 7 or expression vector according to any one of claims 8.
10. the method for producing antibody, comprising:
(i) host cell is converted with nucleic acid molecules as claimed in claim 7 or expression vector according to any one of claims 8,
(ii) in the case where being suitble to the nucleic acid molecules or expression vector is expressed, the host of incubation step (i) described conversion is thin
Born of the same parents, and
(iii) it separates and is purified as expressed by the nucleic acid molecules or expression vector from the host cell that step (ii) is cultivated
Antibody or antibody fragment.
11. pharmaceutical composition includes antibody of any of claims 1-6 or antibody fragment and pharmaceutical acceptable carrier.
12. a kind of method for the HIV-1 infection for detecting human subjects, comprising:
(i) biological sample from the object is made to be in contact with the antibody of any one of claim 1-6 or antibody fragment,
And
(ii) it determines and whether there is the immune complex formed by the antibody or the antibody fragment in the sample,
Wherein there is the immune complex and shows that the object has HIV-1 infection.
13. the purposes of antibody of any of claims 1-6 or antibody fragment, for preventing or treating human subjects
HIV-1 infection.
14. the purposes of antibody according to claim 13 or antibody fragment, wherein the object is lacked with acquired immunity
It falls into syndrome (AIDS).
15. the purposes of antibody according to claim 14 or antibody fragment, wherein antibody or antibody fragment are used for described
The anti-viral pharmaceutical compositions of object application anti-HIV-1.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111944045A (en) * | 2020-08-25 | 2020-11-17 | 贵州医科大学 | anti-HIV antibody or antigen binding fragment thereof, and preparation method and application thereof |
| CN112250762A (en) * | 2020-10-26 | 2021-01-22 | 苏州卫生职业技术学院 | anti-HIV broad-spectrum neutralizing antibody |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130251726A1 (en) * | 2010-09-24 | 2013-09-26 | International Aids Vaccine Initiative | Novel hiv -1 broadly neutralizing antibodies |
| US20150044137A1 (en) * | 2012-03-23 | 2015-02-12 | The United States of America, as represented by the Secretary, Dep. of Health Care Human Services | Neutralizing antibodies to hiv-1 and their use |
| CN107033241A (en) * | 2016-02-03 | 2017-08-11 | 中国疾病预防控制中心性病艾滋病预防控制中心 | HIV-1 wide spectrum neutralizing antibodies and application thereof |
| US20180057570A1 (en) * | 2010-08-31 | 2018-03-01 | Theraclone Sciences, Inc. | Broadly neutralizing human immunodeficiency virus type 1 (hiv-1) gp120-specific monoclonal antibody |
| CN108137676A (en) * | 2015-03-20 | 2018-06-08 | 美国政府(由卫生和人类服务部的部长所代表) | Gp120 neutralizing antibodies and application thereof |
-
2018
- 2018-09-14 CN CN201811074636.5A patent/CN109251246B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20180057570A1 (en) * | 2010-08-31 | 2018-03-01 | Theraclone Sciences, Inc. | Broadly neutralizing human immunodeficiency virus type 1 (hiv-1) gp120-specific monoclonal antibody |
| US20130251726A1 (en) * | 2010-09-24 | 2013-09-26 | International Aids Vaccine Initiative | Novel hiv -1 broadly neutralizing antibodies |
| US20150044137A1 (en) * | 2012-03-23 | 2015-02-12 | The United States of America, as represented by the Secretary, Dep. of Health Care Human Services | Neutralizing antibodies to hiv-1 and their use |
| CN108137676A (en) * | 2015-03-20 | 2018-06-08 | 美国政府(由卫生和人类服务部的部长所代表) | Gp120 neutralizing antibodies and application thereof |
| CN107033241A (en) * | 2016-02-03 | 2017-08-11 | 中国疾病预防控制中心性病艾滋病预防控制中心 | HIV-1 wide spectrum neutralizing antibodies and application thereof |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111944045A (en) * | 2020-08-25 | 2020-11-17 | 贵州医科大学 | anti-HIV antibody or antigen binding fragment thereof, and preparation method and application thereof |
| CN111944045B (en) * | 2020-08-25 | 2021-08-24 | 贵州医科大学 | anti-HIV antibody or antigen binding fragment thereof, and preparation method and application thereof |
| CN112250762A (en) * | 2020-10-26 | 2021-01-22 | 苏州卫生职业技术学院 | anti-HIV broad-spectrum neutralizing antibody |
| CN112250762B (en) * | 2020-10-26 | 2021-06-18 | 苏州卫生职业技术学院 | anti-HIV broad-spectrum neutralizing antibody |
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