阅读说明：本技术 一种多肽以及具有强adcc效应的抗体和应用 (Polypeptide, antibody with strong ADCC effect and application ) 是由 张科 于 2019-11-08 设计创作，主要内容包括：本发明公开了一种多肽以及具有强ADCC效应的抗体和应用,涉及生物医药技术领域。具体地,该多肽的氨基酸序列与SEQ ID No.1-3中的任意一种序列具有至少69％的一致性,该多肽能够作为抗原,用于诱导产生具有强ADCC效应的HIV抗体,能够为有效预防、诊断或治疗HIV提供一种途径。(The invention discloses a polypeptide, an antibody with strong ADCC effect and application, and relates to the technical field of biological medicine. Specifically, the amino acid sequence of the polypeptide has at least 69% of identity with any one of the sequences in SEQ ID No.1-3, the polypeptide can be used as an antigen for inducing the generation of HIV antibodies with strong ADCC effect, and a way for effectively preventing, diagnosing or treating HIV can be provided.)

1. A polypeptide having an amino acid sequence which has at least 69% identity with any one of SEQ ID nos. 1 to 3.

2. The polypeptide of claim 1, wherein the amino acid sequence of the polypeptide is as shown in any one of (a) to (c) below:

(a) a sequence having at least 69% identity to SEQ ID No. 1;

(b) a sequence having at least 80% identity to SEQ ID No. 2;

(c) a sequence having at least 87.5% identity to SEQ ID No. 3;

preferably, the amino acid sequence of the polypeptide is as shown in (d):

(d) as shown in formula 1;

formula 1: ACVPTDP-X1-PQEVVLVNV; in formula 1, X1 is P or N;

preferably, X1 is P.

3. The polypeptide of claim 1 or 2, wherein the amino acid sequence of the polypeptide is as shown in any one of SEQ id nos. 1-3.

4. Use of a polypeptide according to any one of claims 1 to 3 as an antigen for inducing the production of HIV antibodies with a strong ADCC effect.

5. A method for producing an HIV antibody having a strong ADCC effect, comprising: immunizing an animal with the polypeptide of any one of claims 1-3 as an antigen.

6. The method according to claim 5, wherein the animal is selected from one or more of a rabbit, a mouse, an alpaca, a cow, a horse and a camel.

7. An antibody having a strong ADCC effect, which is produced by the method of claim 5 or 6, or an epitope thereof is the polypeptide of any one of claims 1 to 3.

8. An HIV-targeted CAR-T cell having the antibody of claim 7 or an scFV fragment thereof on the surface of its cell membrane.

9. A reagent or a kit for detecting HIV virus, comprising the antibody having strong ADCC effect according to claim 7 or the polypeptide according to any one of claims 1 to 3.

10. A medicament for the treatment of aids comprising an antibody of claim 7 or an HIV-targeted CAR-T cell of claim 8.

Technical Field

The invention relates to the technical field of biological medicines, in particular to a polypeptide, an antibody with strong ADCC effect and application.

Background

AIDS is a very harmful infectious disease caused by infection with the HIV virus. HIV is a virus that attacks the human immune system. It takes the most important CD4T lymphocyte in human immune system as the main target of attack, largely destroys the cell, and makes human body lose immune function.

Therefore, the human body is easy to be infected with various diseases, malignant tumors can occur, and the fatality rate is high. The incubation period of HIV in human body is 8-9 years on average, and before AIDS, the HIV can live and work for many years without any symptoms.

Currently, there is no effective HIV-1 vaccine.

In view of this, the invention is particularly proposed.

Disclosure of Invention

The invention aims to provide a polypeptide, application of the polypeptide as an antigen in inducing generation of an HIV antibody with strong ADCC effect, a method for preparing the HIV antibody with strong ADCC effect, the antibody with strong ADCC effect, a CAR-T cell targeting HIV, a reagent or a kit for detecting HIV virus and a medicine for treating AIDS. The polypeptide can be used as an antigen for inducing the generation of HIV antibodies with strong ADCC effect.

The invention is realized by the following steps:

in a first aspect, the embodiments provide a polypeptide having an amino acid sequence at least 69% identical to any one of SEQ ID nos. 1-3. The polypeptide can be used as an antigen for inducing the generation of HIV antibodies with strong ADCC effect.

In alternative embodiments, the amino acid sequence of the polypeptide is as set forth in any one of (a) - (c) below:

(a) a sequence having at least 69% identity to SEQ ID No. 1;

(b) a sequence having at least 80% identity to SEQ ID No. 2;

(c) a sequence having at least 87.5% identity to SEQ ID No. 3. Sequence conservation of SEQ ID Nos. 1-3 with Induction reference is made specifically to FIG. 1 (based on the results of HIV database analysis, NCBIblast). In FIG. 1, D1 corresponds to the sequence shown by SEQ ID No.1, D2 corresponds to the sequence shown by SEQ ID No.2, and D3 corresponds to the sequence shown by SEQ ID No. 3.

In an alternative embodiment, the amino acid sequence of the polypeptide is as shown in (d):

(d) as shown in formula 1;

formula 1: ACVPTDP-X1-PQEVVLVNV; in formula 1, X1 is P or N;

preferably, X1 is P.

In an alternative embodiment, the amino acid sequence of the polypeptide is as set forth in any one of SEQ ID Nos. 1-3.

In a second aspect, the embodiments of the present invention provide the use of a polypeptide as provided in the above embodiments as an antigen for inducing the production of HIV antibodies with a strong ADCC effect.

In a third aspect, the embodiments of the present invention provide a method for preparing an HIV antibody having a strong ADCC effect, comprising: the polypeptides provided in the above embodiments are used as antigens to immunize animals.

In alternative embodiments, the animal is selected from one or more of a rabbit, a mouse, an alpaca, a cow, a horse and a camel.

In a fourth aspect, the embodiments of the present invention provide an antibody with strong ADCC effect, which is prepared by the method or epitope thereof, which is the polypeptide provided in the above embodiments.

In a fifth aspect, embodiments of the invention provide an HIV-targeted CAR-T cell having the antibody or scFV fragment thereof provided in the above embodiments on the cell membrane surface.

In a sixth aspect, the embodiments of the present invention provide a reagent or a kit for detecting HIV virus, which contains the antibody provided in the above embodiments or the polypeptide provided in any of the above embodiments.

In a seventh aspect, the embodiments of the present invention provide a medicament for treating aids, which comprises the antibody provided in the above embodiments or the CAR-T cell provided in the above embodiments.

The invention has the following beneficial effects:

the application provides a polypeptide, the amino acid sequence of the polypeptide has at least 69% of identity with any one sequence in SEQ ID No.1-3, the polypeptide can be used as an antigen for inducing the generation of HIV antibodies with strong ADCC effect, and a way for effectively preventing, diagnosing or treating HIV is provided.

In addition, the embodiment of the invention also provides application of the polypeptide as an antigen in inducing generation of an HIV antibody with strong ADCC effect, a method for preparing the HIV antibody with strong ADCC effect, the antibody with strong ADCC effect, a CAR-T cell targeting HIV, a reagent or a kit for detecting HIV virus and a medicine for treating AIDS.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.

FIG. 1 shows the conservation of the sequences shown in SEQ ID No.1 to SEQ ID No.3 in the summary of the present invention;

FIG. 2 is a flow chart of the construction of a yeast-displayed HIV-1HXB2 fragment library as provided in example 4 of the present invention;

FIG. 3 shows the result 1 of the measurement of recombinant yeast cells by the flow cytometer provided in example 4 of the present invention;

FIG. 4 shows the result 2 of the measurement of recombinant yeast cells by the flow cytometer provided in example 4 of the present invention;

FIG. 5 shows the measurement result 3 of recombinant yeast cells by the flow cytometer provided in example 4 of the present invention;

FIG. 6 is a depiction of the IgG sequence of a positive yeast clone provided in example 4 of the present invention used to map the epitope to full length HIV-1HXB2 gp 160;

FIG. 7 shows the results of experiments for eliminating the antibody absorption of the epitope provided in test example 1 of the present invention;

FIG. 8 shows the results of screening for memory B cells using epitopes provided in test example 2 of the present invention.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.

The features and properties of the present invention are described in further detail below with reference to examples.