CN108929382B

CN108929382B - The antibody of anti-SERINC5 a kind of and its application

Info

Publication number: CN108929382B
Application number: CN201810800332.6A
Authority: CN
Inventors: 程林; 唐娴; 王辉; 陈志伟; 何云
Original assignee: Third Peoples Hospital of Shenzhen
Current assignee: Third Peoples Hospital of Shenzhen
Priority date: 2018-07-20
Filing date: 2018-07-20
Publication date: 2019-10-15
Anticipated expiration: 2038-07-20
Also published as: CN108929382A

Abstract

The present invention relates to the antibody of anti-SERINC5 a kind of and its applications, specifically, the present invention provides the antibody of anti-SERINC5 a kind of and functional fragment, nucleotide, carrier, monoclonal antibody, kit and its applications in the polypeptide sequence shown in detection SERINC5 albumen or SEQ ID NO:9, meanwhile the present invention provides a kind of methods for preparing monoclonal antibody, the method for preparing monoclonal antibody hybridoma cell line.Using the antibody and functional fragment of the method preparation in the present invention can be effectively used for in the reacting of SERINC5 albumen, the SERINC5 such as expressed in the SERINC5 molecule of Flow cytometry cell surface, combination cell, study virus amplification mechanism etc., this will provide effective research tool for the antiviral-mechanism for studying SERINC5, it will be helpful to accelerate the progress of this field, the novel AIDS-treating medicine for further exploitation targeting HIV albumen Nef provides new thinking.

Description

The antibody of anti-SERINC5 a kind of and its application

Technical field

The present invention relates to molecular biology fields, and in particular to the antibody of anti-SERINC5 a kind of and its application.

Background technique

Serine integration factor (SERINC) is a kind of memebrane protein, and family shares 5 members, and wherein SERINC5 is mesh The innate immune molecule of the AIDS virus resisting of preceding latest find, can blocking virus infect new cell, to significantly reduce people The infection ability of para-immunity defective virus (HIV).Influence of the SERINC5 albumen to viral infectivity is very big, can be by inhibition of HIV grain The contagiousness of son reduces by more than 100 times.HIV encodes a kind of regulatory protein Nef to resist SERINC5.HIV inhibits SERINC albumen Ability have a strong influence for cell of its infection, destroy and this mechanism of SERINC inhibited to may make SERINC5 egg It is white can activated virus, be treat HIV and express Nef albumen similar virus a kind of very powerful strategy.

Other than being able to suppress HIV, recent studies have found that, the antiretroviral effect of SERINC5 albumen seems horizontal Across all retrovirus, it may be possible to generally existing.SERINC5 albumen inhibit all retrovirus, even with The contagiousness of the farthest retrovirus of HIV-1 affiliation.The function and its mechanism for furtheing investigate SERINC5 albumen are possible to The treatment of all envelope virus can be had an impact.

However, the antiviral activity of each member of SERINC family, internal expression and antiviral-mechanism, scientific research personnel also know It is few.It it is not immediately clear that SERINC5 albumen prevents virion from downloading to its genome and goes in new host cell Precise mechanism.One kind may be that SERINC5 physically prevents viral genome to enter by peplos and cell membrane In new host cell.For this process and SERINC operating mechanism we still there are many thing is unclear.And SERINC5 Monoclonal antibody specific can be especially useful for the shortage of the monoclonal antibody of Flow cytometry, hinder researcher Disclose the research progress of SERINC5 antiviral functions.

Summary of the invention

The present invention is directed to solve in the related technology at least to a certain extent, for this purpose, the purpose of the present invention It is to provide a kind of SERINC5 monoclonal antibody specific or functional fragment, the antibody or functional fragment can be with SERINC5 It effectively combines, so as to the mechanism of action for studying SERINC5, this will be provided with for the antiviral-mechanism for studying SERINC5 The research tool of effect, it will help accelerate the progress of this field, for the novel Chinese mugwort of further exploitation targeting HIV albumen Nef It grows medicine and new thinking is provided.

For this purpose, the first aspect of the present invention provides a kind of antibody or functional fragment, it is selected from:

A) antibody or functional fragment, comprising amino acid sequence shown in SEQ ID NO:1-SEQ ID NO:3, and/or Amino acid sequence shown in SEQ ID NO:4-SEQ ID NO:6；

B) antibody or functional fragment, containing respectively with SEQ ID NO:1-SEQ ID NO:3 and/or SEQ ID NO: Amino acid sequence homology sequence shown in 4-SEQ ID NO:6 wherein at least has a sequence to have and SEQ ID NO: 1-SEQ ID NO:6 is not less than 95%, 96%, 97%, 98% or 99% homology；

C) a)-b) described in the modification through amino acid residue of antibody or functional fragment, replace, miss or add and formed Amino acid derived sequence.

Antibody or functional fragment recited above can be in conjunction with SERINC5.

Wherein amino acid sequence shown in SEQ ID NO:1-SEQ ID NO:3 is respectively complementary determining region of heavy chain CDR1- Amino acid sequence shown in CDR3, SEQ ID NO:4-SEQ ID NO:6 is respectively complementary determining region of light chain CDR1-CDR3.

In a preferred example, the antibody or functional fragment contain SEQ ID NO:10 and/or SEQ ID NO:11 Shown in amino acid sequence.

In a preferred example, the antibody or functional fragment are source of mouse antibody or functional fragment.

The second aspect of the present invention provides a kind of nucleotide, and the nucleotide sequence contains encoding such antibodies or function The sequence of property segment；

In a preferred example, the nucleotide sequence includes sequence shown in SEQ ID NO:7 and/or SEQ ID NO:8 Column.

The third aspect of the present invention provides a kind of carrier, and the carrier contains above-mentioned nucleotide sequence.

The fourth aspect of the present invention provides a kind of monoclonal antibody, and the monoclonal antibody is by hybridoma cell strain 4D4.8, deposit number are CCTCC NO:C2018140 expression.

The fifth aspect of the present invention provide it is a kind of prepare hybridoma cell strain 4D4.8, deposit number is CCTCC NO: The method of C2018140, described method includes following steps:

Mouse is immunized in reagent containing polypeptide sequence shown in SEQ ID NO:9；

Take the spleen cell and SP2/0 cell fusion of immunized mice；

The fused cell that can be reacted with SERINC5 albumen by immune detection reaction screening.

The sixth aspect of the present invention provides a kind of method for preparing monoclonal antibody, i.e., cultivates in suitable culture medium Hybridoma cell strain 4D4.8, deposit number are CCTCC NO:C2018140.

The seventh aspect of the present invention provides a kind of kit, and the kit includes above-mentioned antibody or functional piece Section or nucleotide or carrier or monoclonal antibody.

The seventh aspect of the present invention provides antibody or functional fragment or nucleotide or carrier or monoclonal antibody exists Detect the application in polypeptide sequence shown in SERINC5 albumen or SEQ ID NO:9.

The detection, which is not limited to polypeptide sequence shown in detection SERINC5 albumen or SEQ ID NO:9, whether there is, also Including screening, target protein or polypeptide sequence specific bond is made to carry out other researchs etc., it is e.g., and more shown in SEQ ID NO:9 Peptide is combined derived from the immunogene polypeptid specificity of SERINC5ECL4；Western blot detects the expression of SERINC5；Using In the SERINC5 molecule of Flow cytometry cell surface；The SERINC5 of combination cell expression, studies virus amplification mechanism Application in etc..

Beneficial effect obtained by the present invention

The present invention provides a kind of antibody and functional fragments, can effectively be detected using the antibody and functional fragment The peptide molecule with immunogenicity of SERINC5 albumen and source SERINC5 albumen, such antibody can be effectively used for In the reaction of SERINC5 albumen, as being expressed the SERINC5 molecule of Flow cytometry cell surface, combination cell SERINC5, research virus amplification mechanism etc., this will provide effective research tool for the antiviral-mechanism for studying SERINC5, will The progress of this field is helped speed up, is provided for the novel AIDS-treating medicine that further exploitation targets HIV albumen Nef new Thinking.

Word paraphrase:

Functional fragment: structure is in a kind of compound between amino acid and protein, has bioactivity.

Detailed description of the invention

Fig. 1 is strong and weak for the amino acid sequence and its hydrophily of SERINC5 albumen extracellular ring 4, immunogenicity and surface probability, And the polypeptide sequence (box mark) for Antibody preparation selected.

Fig. 2 is that ELISA filters out positive colony 4D4 and 5H12.

Fig. 3 is the result figure for the combination that ELISA detects 4D4.6,4D4.8,4D4.11,5H4.1 and polypeptide.

Fig. 4 is that flow cytometry monoclonal antibody 4D4.6,4D4.8,4D4.11,5H4.1 and 293T-SERINC5 are thin The result figure of the combination of born of the same parents, grey peak is hybridoma culture supernatant and 293T cell effect as a result, the peak of white is training in figure Support the result of supernatant and 293T-SERINC5 cell effect.

Fig. 5 is the table that Flow cytometry monoclonal antibody 4D4.8 and 5H4.1 detects PBMC cell surface SERINC5 The result figure reached.

Fig. 6 is the result figure for the expression that Western blot monoclonal antibody 4D4.8 detects SERINC5.

Fig. 7 is monoclonal antibody 4D4.8 Subtype result.

Fig. 8 is 4D4.8 weight chain variabl area sequence structure chart.

Fig. 9 is 4D4.8 light-chain variable sequence structure chart.

Biomaterial preservation:

Hybridoma cell strain send China typical culture collection center preservation in the present invention, and deposit number is CCTCC NO: C2018140, address are located at Wuhan, China university, and preservation date is on June 28th, 2018, and classification naming is hybridoma Strain 4D4.8.

Specific embodiment

The present invention provides the monoclonal antibody specific that can resist SERINC5 and its functional fragments, encoding antibody Nucleotide sequence, carrier and its application.

Under the premise of not substantial effect antibody activity, those skilled in the art can to sequence of the invention replace, One or more (such as 1,2,3,4,5,6,7,8,9 or 10 or more) amino acid are added and/or lack, to obtain State the variant of antibody or the sequence of its functional fragment.They are considered as including within the protection scope of the present invention.Such as can Become area to be replaced the amino acid with similarity.The sequence of variant of the present invention can have at least with its derived sequences There is 95%, 96%, 97%, 98% or 99% consistency.It is soft that sequence analysis can be used in sequence identity of the present invention Part measurement.Such as the computer program BLAST using default parameter, especially BLASTP or TBLASTN.

Antibody of the invention can be (for example, IgM, IgG1 or IgG4 antibody) of overall length or can only include antigen-binding portion Divide (for example, Fab, F (ab ') 2 or scFv segment), or can be modified to influence function.The present invention includes the sugar with modification The anti-SERINC5 antibody of base mode.In some applications, it is modified to remove undesirable glycosylation site and can be , or on oligonucleotide chain there is no fucose moieties for example to enhance the anti-of antibody-dependent cytotoxicity (ADCC) function Body.In other applications, galactosylation modification can be carried out to change complement-dependent cytotoxicity (CDC).

Term as used herein " functional fragment " is referred to combining active segment with SERINC5, be referred in particular to Antibody fragment such as Fv, scFv (sc refers to single-stranded), Fab, F (ab ') 2, Fab ', scFv-Fc segment or double antibody (diabody), Or by chemical modification or any segment by that should be able to increase half life in incorporation liposome, the chemical modification is for example Add poly- (alkylidene) glycol such as polyethylene glycol (" Pegylation, PEGylated ") (referred to as Fv-PEG, scFv-PEG, Fab- The pegylated fragments of PEG, F (ab ') 2-PEG or Fab '-PEG) (" PEG " is polyethylene glycol).Preferably, the functional sheet Section will be made of the partial sequence of the weight of its derived antibodies or light variable chains or comprising them, the partial sequence is enough to retain Binding specificity identical with its derived antibodies and sufficient affinity, for SERINC5, preferably at least equal to its derived antibodies The 1/100 of affinity, at least equal to 1/10 in more preferable mode.This function fragment will include minimum 5 amino acid, preferably 10,15,25,50 and 100 continuous amino acids of the antibody sequence in its source.

PBS:NaCl 137mmol/L, KCl 2.7mmol/L, Na2HPO4 10mmol/L, KH2PO4 2mmol/L

Western blot sample-loading buffer: it is purchased from the green skies (article No.: P0015)

Below with reference to specific embodiments and the drawings, the present invention will be described.It should be noted that these embodiments are only It is illustrative, and is not considered as limiting the invention.Particular technique or condition are not specified in embodiment, according to ability Technology or conditions described in document in domain are carried out according to product description.Factory is not specified in agents useful for same or instrument Shang Zhe, being can be with conventional products that are commercially available.

The screening of the polypeptide immunogen of 1 SERINC5 of embodiment

SERINC5 (GENBANK ACCESSION:NM_001174072) is 9 transmembrane proteins, is not possible to prepare at present pure Change albumen, therefore use polypeptide that mouse is immunized, prepares the strategy of monoclonal antibody.

SERINC5 extracellular region includes aminoterminal and 4 extracellular rings, and wherein aminoterminal includes 35 amino acid；1st extracellular Ring includes 14 amino acid residues, and contains 1 glycosylation site；2nd extracellular ring includes 21 amino acid residues, contains 1 A glycosylation site；3rd extracellular ring only includes 7 amino acid；4th extracellular ring is made of 53 amino acid residues, and nothing Glycosylation site.The aminoterminal of SERINC5 and the 4th extracellular ring region are more comprising amino acid residue, and aglycosylated site, than It is easier to the polypeptide filtered out He Shi as immunogene.Find that the region of aminoterminal C25-R35 is exempted from by DNASTAR software analysis Epidemic focus is stronger, but due to shorter only 10 amino acid in the region, and abuts cell membrane, has space steric effect, is not reason The immunogene thought.G362-G375 peptide fragment in the 4th extracellular ring region has good immunogenicity, hydrophily and table Face frequency (Fig. 1), is suitable as immunogene, therefore synthesis polypeptide and is coupled keyhole limpet hemocyanin (KLH), mouse, preparation is immunized Monoclonal antibody.

Synthetic peptide sequence is as follows:

GGEDTEEQQPGKEG (SEQ ID NO:9),

The synthesis of aforementioned polypeptides and complete by Nanjing Genscript Biotechnology Co., Ltd. with the coupling of KLH (hemocyanin) At.

The preparation of the anti-human SERINC5 monoclonal antibody of embodiment 2

1. immune animal

It is 1mg/ml as immunogene that distilled water, which dissolves aforementioned polypeptides (the SEQ ID NO:9) compound concentration of KLH coupling, is taken Immunogene mixes concussion with each 100 μ l of Freund's complete adjuvant (Sigma-Aldrich) and forms water in oil emulsion.Initial immunity takes 6-8 week old BALB/c mouse carries out multiple spot and 200 μ l immunogenes/adjuvant mixed liquor is subcutaneously injected.After initial immunity, respectively at The booster immunization for once halving dosage is carried out within 3rd week and the 6th week, and was taken a blood sample with the 8th week, elisa assay antibody titer.

2. cell fusion

2 weeks after last time booster immunization, mouse is put to death, spleen cell is taken, according to the ratio and SP2/0 cell of 10:1 (being purchased from ATCC, article No.: CRL-1581) mixing, carries out cell with PEG1450 (Sigma-Aldrich) according to a conventional method after centrifugation Fusion.Fused cell is resuspended in containing HAT (hypoxanthine (h), aminopterin (a), thymidine (t)) and 20%FBS (tire ox blood (article No.: Gibco in 10569010), is inoculated with 96 porocyte culture plates, is placed in 5%CO DMEM screening and culturing medium clearly)₂, 37 DEG C Under the conditions of cultivate.

3. filtering hybridoma

Multiple Hybridoma Cell Cultures 10~12 days will prepared in 2, when hybridoma is paved with 1/10 area of bottom hole, 100 μ l culture supernatants are taken, are added separately to the naked peptide of the polypeptide synthesized in embodiment 1 (SEQ ID NO:9), with the dense of 10 μ g/ml Spend coated elisa plate in, with conventional ELISA method carries out positive colony screening (HRP mark secondary antibody be purchased from the Divine Land Beijing Yi Qiao Science and Technology Ltd., article No.: SSA007-200), as a result, it has been found that the hole D4 of the 4th piece of ELISA Plate is in strong positive, it is named as The H4 of 4D4, the 5th piece of ELISA Plate are in weakly positive, are named as 5H4 (Fig. 2).When hybridoma is paved with about 20% area of bottom hole When, positive colony 4D4 is subcloned with limiting dilution assay, by above-mentioned ELISA method, filter out positive colony 4D4.6, 4D4.8,4D4.11,5H4.1 (Fig. 3).

3 Flow cytometry cell surface SERINC5 of embodiment

Each 100 μ l of the culture supernatant of hybridoma 4D4.6,4D4.8,4D4.11,5H4.1 that Example 2 filters out, Respectively with 1 × 10⁶ A 293T cell and 293T-SERINC5 cell (the 293T cell line for stablizing expression people SERINC5) (above-mentioned 2 A cell line is the preservation of The Third People's Hospital of Shenzhen's hepatopathy research institute) it carries out being incubated for 10 minutes at room temperature, add PBS centrifugation to wash It washs, adds 0.2 μ l fluorescence secondary antibody AF647-Anti mouse IgG (H+L) (Invitrogen, article No. A-21237), room temperature is protected from light It is incubated for 10 minutes, after PBS washing, Flow cytometry cell surface fluorescence signal.As a result, it has been found that 4D4.6,4D4.8, 4D4.11 can identify the SERINC5 molecule of cell surface, and 5H4.1 can only Recognition polypeptide, cannot identify cell surface (Fig. 4, grey peak is hybridoma culture supernatant and 293T cell effect as a result, the peak of white is culture to SERINC5 molecule in figure The result of supernatant and 293T-SERINC5 cell effect).

To detect cd4 t cell SERINC5 developed by molecule situation, 100 μ l of Healthy People periphery anticoagulation is taken, is separately added into 100 μ l hybridoma 4D4.8,5H4.1 (negative control) culture supernatant, 5 μ l PerCP/Cy5.5anti-human CD3 (PerCP.Cy5.5CD3) (Biolegend, article No. 317336), 5 μ l FITC anti-human CD4 (FITC.CD4) (Biolegend, article No. 357406) mixes incubation at room temperature 10 minutes, after 2ml PBS washing is added, adds 0.2 μ l fluorescence secondary antibody AF647-Anti mouse IgM (Invitrogen, article No. M31601), room temperature are protected from light incubation 10 minutes, and 2ml red blood cell is added Lysate (the green skies, article No. C3702) is incubated at room temperature 5 minutes splitting erythrocytes, centrifugation, Flow cytometry after PBS washing Cell surface fluorescence signal.Lymphocyte (Fig. 5 A) is chosen by FSC and SSC further to analyze, and passes through the expression of CD3 and CD4 Lymphocyte is divided into 3 groups by situation, choose double positive CD4T lymphocytes (Fig. 5 B) further analysis 4D4.8 and 5H4.1 with The combination situation of cell surface SERINC5.As a result the same similar to the testing result of 293T-SERINC5,4D4.8 can detecte The SERINC5 on cd4 t cell surface, and 5H4.1 then cannot.The above result shows that can be used for streaming thin by monoclonal antibody 4D4.8 Born of the same parents' art detects cell surface SERINC5 molecule (Fig. 5 C-D).

The expression of 4 Western blot of embodiment detection cell SERINC5

Take 4 × 10⁶Healthy human peripheral blood mononuclearcell (PBMC) and 1 × 10⁶293T cell, is conventionally extracted Whole-cell protein, by 4:1 volume ratio be added 5 × Western blot sample-loading buffer, 100 DEG C processing sample 10 minutes, respectively 15 μ l samples are taken, SDS-PAGE, transferring film, closing are conventionally run, then use 5ml hybridoma 4D4.8 culture supernatant room temperature After being incubated for 1 hour, conventional method washed, the secondary antibody of HRP label (purchased from the Divine Land Beijing Yi Qiao Science and Technology Ltd., article No.: SSA007-200 it) is incubated for, the exposure of ECL reagent, the band of SERINC5 is as a result found in expected position, shows monoclonal antibody 4D4.8 can be used for the expression (Fig. 6) of Western blot experiment detection SERINC5.

5 Subtype of embodiment

Using

Rapid Isotyping Kits-Mouse (Thermo Scientific) mouse monoclonal hypotype Assay kit analyzes the hypotype of monoclonal antibody 4D4.8 according to kit specification, and 4D4.8 is anti-for IgM class as the result is shown Body, light chain are κ chain (Fig. 7).

6 antibody variable sequences of embodiment

By hybridoma 4D4.8 total quantity culture to 10⁷A cell, 1000rpm 10 minutes collection cells of centrifugation, and with Trizol kit (Invitrogen, article No. 15596026) extracts total serum IgE according to kit specification.Using the RNA as mould Plate synthesizes the first chain cDNA (TAKARA, article No. 6110A), right for subsequent template amplification hybridoma with the first chain cDNA The variable region DNA sequence dna answered.Primer sequence used in amplified reaction and the first framework region of antibody variable region and constant region are mutual Mend, see Table 1 for details for primer sequence, in 50 μ l reaction systems, be separately added into 1 μ l, 10 × PCR buffer of cDNA, 5 μ l, upstream and Each 1 μ l (25pmol) of downstream primer, 1 μ l of dNTP, Taq enzyme (Invitrogen) 1 μ l, H₂O 40 μ l, 95 DEG C of initial denaturation 5min, Into temperature cycles, PCR amplification is carried out.Reaction condition is 94 DEG C of denaturation 1min, and 55 DEG C of annealing 1min, 72 DEG C extend 30 seconds, altogether 30 recycle, then 72 DEG C of heat preservation 10min, 4 DEG C of coolings.

After amplified production is sequenced, hybridoma clone 4D4.8 weight chain variabl area sequence structure (Fig. 8) and light chain variable are obtained Region sequence structure (Fig. 9) and its corresponding nucleotide sequence, particular sequence are as shown in table 2:

Table 1: primer sequence

K, Y, S, W and R are respectively degeneracy base in table 1, and wherein K corresponds to G/T, and Y corresponds to C/T, and S corresponds to G/C, and W corresponding A/ T, R corresponding A/G.

Table 2:4D4.8 heavy chain and sequence of light chain

In the description of this explanation, reference term " one embodiment ", " some embodiments ", " example ", " specific example ", Or the description of " some examples " etc. means particular features, structures, materials, or characteristics packet described in conjunction with this embodiment or example Contained at least one embodiment of the present invention or example.In the present specification, schematic expression of the above terms need not needle Pair be identical embodiment or example.Moreover, particular features, structures, materials, or characteristics described can any one or It can be combined in any suitable manner in multiple embodiment or examples.In addition, without conflicting with each other, those skilled in the art The feature of different embodiments or examples described in this specification and different embodiments or examples can be combined and group It closes.

Although the embodiments of the present invention has been shown and described above, it is to be understood that above-described embodiment is example Property, it is not considered as limiting the invention, those skilled in the art within the scope of the invention can be to above-mentioned Embodiment is changed, modifies, replacement and variant.

Sequence table

<110>The Third People's Hospital of Shenzhen

<120>antibody of anti-SERINC5 a kind of and its application

<130> 20180702

<160> 28

<170> PatentIn version 3.5

<210> 1

<211> 8

<212> PRT

<213> Mus musculus

<400> 1

Gly Tyr Thr Phe Thr Ser Tyr Trp

1 5

<210> 2

<211> 8

<212> PRT

<213> Mus musculus

<400> 2

Ile Tyr Pro Gly Asp Gly Asp Thr

1 5

<210> 3

<211> 14

<212> PRT

<213> Mus musculus

<400> 3

Ala Arg Ser Thr Tyr Gly Ser Ser Phe Ala Trp Phe Ala Tyr

1 5 10

<210> 4

<211> 11

<212> PRT

<213> Mus musculus

<400> 4

Gln Ser Ile Val His Ser Asn Gly Asn Thr Tyr

1 5 10

<210> 5

<211> 3

<212> PRT

<213> Mus musculus

<400> 5

Lys Val Ser

1

<210> 6

<211> 9

<212> PRT

<213> Mus musculus

<400> 6

Phe Gln Gly Ser His Val Pro Trp Thr

1 5

<210> 7

<211> 363

<212> DNA

<213> Mus musculus

<400> 7

caggttcagc tccagcagtc tggggctgag ctggcaagac ctggggcttc agtgaagttg 60

tcctgcaagg cttctggcta cacctttact agctactgga tgcagtgggt aaaacagagg 120

cctggacagg gtctggaatg gattggggct atttatcctg gagatggtga tactaggtac 180

actcagaagt tcaagggcaa ggccacattg actgcagata aatcctccag cacagcctac 240

atgcaactca gcagcttggc atctgaggac tctgcggtct attactgtgc aagatcgacg 300

tacggtagta gcttcgcctg gtttgcttac tggggccaag ggactctggt cactgtctct 360

gca 363

<210> 8

<211> 342

<212> DNA

<213> Mus musculus

<400> 8

gatgttttga tgacccaaac tccactctcc ctgcctgtca gtcttggaga tcaagcctcc 60

atctcttgca gatctagtca gagcattgta catagtaatg gaaacaccta tttagaatgg 120

tacctgcaga aaccaggcca gtctccaaag ctcctgatct acaaagtttc caaccgattt 180

tctggggtcc cagacaggtt cagtggcagt ggatcaggga cagatttcac actcaagatc 240

agcagagtgg aggctgagga tctgggagtt tattactgct ttcaaggttc acatgttccg 300

tggacgttcg gtggaggcac caagctggaa atcaaagctgat 342

<210> 9

<211> 14

<212> PRT

<213>artificial sequence

<400> 9

Gly Gly Glu Asp Thr Glu Glu Gln Gln Pro Gly Lys Glu Gly

1 5 10

<210> 10

<211> 121

<212> PRT

<213> Mus musculus

<400> 10

Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met Gln Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Ala Ile Tyr Pro Gly Asp Gly Asp Thr Arg Tyr Thr Gln Lys Phe

50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ser Thr Tyr Gly Ser Ser Phe Ala Trp Phe Ala Tyr Trp Gly

100 105 110

Gln Gly Thr Leu Val Thr Val Ser Ala

115 120

<210> 11

<211> 114

<212> PRT

<213> Mus musculus

<400> 11

Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly

1 5 10 15

Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser

20 25 30

Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly

85 90 95

Ser His Val Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

Ala Asp

<210> 12

<211> 21

<212> DNA

<213>artificial sequence

<400> 12

ggatacagtt ggtgcagcat c 21

<210> 13

<211> 21

<212> DNA

<213>artificial sequence

<400> 13

gatgttttga tgacccaaac t 21

<210> 14

<211> 21

<212> DNA

<213>artificial sequence

<400> 14

gatattgtga tgacgcaggc t 21

<210> 15

<211> 18

<212> DNA

<213>artificial sequence

<400> 15

gatattgtga taacccag 18

<210> 16

<211> 21

<212> DNA

<213>artificial sequence

<400> 16

gacattgtgc tgacccaatc t 21

<210> 17

<211> 21

<212> DNA

<213>artificial sequence

<400> 17

gacattgtga tgacccagtc t 21

<210> 18

<211> 21

<212> DNA

<213>artificial sequence

<400> 18

gatattgtgc taactcagtc t 21

<210> 19

<211> 21

<212> DNA

<213>artificial sequence

<400> 19

gatatccaga tgacacagac t 21

<210> 20

<211> 21

<212> DNA

<213>artificial sequence

<400> 20

gacatccagc tgactcagtc t 21

<210> 21

<211> 21

<212> DNA

<213>artificial sequence

<400> 21

caaattgttc tcacccagtc t 21

<210> 22

<211> 21

<212> DNA

<213>artificial sequence

<400> 22

gacattctga tgacccagtc t 21

<210> 23

<211> 21

<212> DNA

<213>artificial sequence

<400> 23

atggtgytaa ktcttytgta s 21

<210> 24

<211> 24

<212> DNA

<213>artificial sequence

<400> 24

atggctgtyy tggtgctgtt cctc 24

<210> 25

<211> 22

<212> DNA

<213>artificial sequence

<400> 25

atgggawgga gctgkrtctt yc 22

<210> 26

<211> 24

<212> DNA

<213>artificial sequence

<400> 26

atgaartkca gctgggtcat sttc 24

<210> 27

<211> 22

<212> DNA

<213>artificial sequence

<400> 27

atgaagttgt tggstgaact gg 22

<210> 28

<211> 19

<212> DNA

<213>artificial sequence

<400> 28

acatttggga aggactgac 19

Claims

1. a kind of antibody or functional fragment, which is characterized in that

It is SEQ ID NO:1, SEQ ID NO:2, SEQ ID that HCDR1, HCDR2, HCDR3 of its heavy chain variable region, which are respectively corresponded, Amino acid sequence shown in NO:3, and it is SEQ ID NO:4, SEQ that LCDR1, LCDR2, LCDR3 of its light chain variable region, which are respectively corresponded, Amino acid sequence shown in ID NO:5, SEQ ID NO:6；

The antibody or functional fragment can be in conjunction with SERINC5.

2. antibody or functional fragment according to claim 1, it is characterized in that, the heavy chain of the antibody or functional fragment Variable region is amino acid sequence shown in SEQ ID NO:10, and light chain variable region is amino acid sequence shown in SEQ ID NO:11 Column.

3. antibody or functional fragment according to claim 2, it is characterized in that, the antibody or function fragment are source of mouse antibody Or function fragment.

4. a kind of polynucleotides, it is characterized in that, the polynucleotide sequence contain coding antibody of any of claims 1 or 2 or The sequence of functional fragment.

5. polynucleotides according to claim 4, it is characterized in that, the polynucleotide sequence include SEQ ID NO:7 and Sequence shown in SEQ ID NO:8.

6. a kind of carrier, it is characterized in that, the carrier contains polynucleotide sequence as claimed in claim 4.

7. a kind of monoclonal antibody, it is characterized in that, the monoclonal antibody is the miscellaneous of CCTCC NO:C2018140 by deposit number Hand over tumor cell strain 4D4.8 expression.

8. monoclonal antibody according to claim 7, it is characterized in that, the monoclonal antibody is IgM antibody.

9. a kind of kit, it is characterized in that, the kit includes antibody of any of claims 1-3 or functionality Polynucleotides described in any one of segment or claim 4-5 or carrier as claimed in claim 6 or claim 7 institute The monoclonal antibody stated.

10. described in any one of antibody of any of claims 1-3 or functional fragment or claim 4-5 Polynucleotides or carrier as claimed in claim 6 or monoclonal antibody as claimed in claim 7 in detection SERINC5 albumen or Application in polypeptide sequence shown in SEQ ID NO:9.