CN1108819C - Preparation of dissociation-resisting epitope vaccine - Google Patents

Abstract

The invention belongs to the technical field of bioengineering. The invention first artificially synthesizes epitope polypeptides of several restricted mutant epitopes on one or more specific key epitopes on one or more antigenic proteins; or repeated forms appear on a synthetic polypeptide; the epitope polypeptide is coupled to the carrier protein or carrier polypeptide or directly synthesized into a dendritic molecule using the MAP technology system; the conjugate of the epitope polypeptide is prepared with an appropriate adjuvant into an anti-variant-epitope vaccine. The invention can induce epitope-specific immune response; can deal with vaccine failure caused by pathogen variation; and can prepare multiple vaccines.

Description

Preparation method of anti-mutation-epitope vaccine

The invention belongs to the technical field of bioengineering, and particularly relates to the preparation and production of epitope vaccines.

Conventional vaccine preparation techniques usually use the following methods:

(1) Utilize attenuated or inactivated pathogens (such as poliovirus);

(2) Subunit vaccines prepared using natural or genetically recombined components of pathogens, such as bacterial toxoids, viral envelope proteins, etc.;

(3) The nucleic acid of the pathogen is used to prepare a vaccine.

These existing vaccine preparation technologies have great difficulties in preventing infectious diseases caused by certain pathogens (such as AIDS virus and influenza virus) with high mutation frequency; in addition, existing vaccines cannot directly induce predetermined epitope-specific immune response.

The purpose of the present invention is to solve or partly solve the worldwide problem of vaccine failure caused by pathogen mutation, and design a preparation method of anti-mutation-epitope vaccine, which has the pathogen or pathogen that does not need to be attenuated or inactivated. Natural protein or genetically recombinant protein antigens and nucleic acids of pathogens; can directly induce a predetermined epitope-specific immune response; can deal with vaccine failure caused by pathogen variation; can prepare multiple vaccines against different pathogens, etc. Advantage.

The preparation method of the anti-mutation-epitope vaccine that the present invention proposes is characterized in that comprising the following steps:

1) Artificially synthesizing epitope polypeptides of several restricted mutant epitopes on one or more specific key epitopes on one or more antigenic proteins;

2) The one or more mutant epitopes appear in one or repeated forms on a synthetic polypeptide (ie, multiple mutant epitope-epitope polypeptides);

3) The epitope polypeptide is coupled to a carrier protein or carrier polypeptide or directly synthesized into a dendritic molecule using the MAP (Multiple AntigenPeptide) technology system;

4) The epitope polypeptide conjugates or dendrimers are prepared with appropriate adjuvants to prepare anti-mutation-epitope vaccine or anti-mutation-multi-epitope-epitope vaccine.

The preparation method of the epitope vaccine designed by the present invention according to the brand-new theory (Immunology Today, 20:588-589, December 1999) of the epitope vaccine firstly proposed by the inventor has the following effects:

First, it can induce a strong immune response against a specific epitope, and can significantly increase the level of antibodies against a specific epitope (10-20 times) compared with the corresponding subunit vaccine;

Second, specific epitopes on some natural proteins cannot induce a strong immune response with protective effect in the body, and the vaccine prepared by the present invention can induce high-titer epitope-specific antibodies (epitope The amount of specific antibodies is 10-240 micrograms per milliliter of serum), which can play an effective role in immune protection;

Third, the anti-mutation-multi-epitope-epitope vaccine prepared by the present invention can simultaneously induce antibody responses to multiple restricted mutant variant epitopes in the body.

Embodiment 1: Preparation of HIV-1 anti-mutation-multi-epitope-epitope vaccine

Step 1. Synthesis of two anti-mutation-epitope polypeptides containing four and three restriction mutants, respectively

CG(ELDKWA)G(ELEKWA)G(ELNKWA)G(ELDNWA)

CG(GPGRAFY)G(GPGKAFY)G(GPGQAFY)

Step 2. Utilize MBS (m-maleimidobenzoyl-N-hydroxy succinimide ester) to combine two kinds of anti-mutation epitope polypeptides

After mixing, couple with the carrier protein BSA, or couple with BSA separately and mix;

Step 3. Prepare the anti-mutation-epitope vaccine by mixing the conjugate with the adjuvant respectively.

Example 2: Preparation of HIV-1 Anti-Variation-Epitope Vaccine Using Dendritic Epitope Polypeptide Molecules

Step 1. Direct synthesis using MAP (Multiple Antigen Peptide) technology system

On the dendrimers of four defined mutants containing a specific epitope ELDKWA;

Step 2. Mix the synthetic dendritic epitope polypeptide molecules with appropriate adjuvants to prepare anti-mutation-epitope vaccines

Seedling.

Claims (2)

1, a kind of preparation method of dissociation-resisting epitope vaccine is characterized in that may further comprise the steps:

1) epitope polypeptide of the several restrictive mutation type epi-positions on one or more the specific critical epitopes on one or more antigen proteins of synthetic;

2) said or a plurality of saltant epi-positions are with one or appear at multiple form on the polypeptide of synthetic;

3) said epitope polypeptide is coupled to carrier protein or carrier polypeptide;

4) coupling matter of said epitope polypeptide is equipped with that proper adjuvant is prepared into dissociation-resisting epitope vaccine or resistance is different-multiepitope epitope vaccine.

2, a kind of preparation method of dissociation-resisting epitope vaccine is characterized in that may further comprise the steps:

1) epitope polypeptide of the several restrictive mutation type epi-positions on one or more the specific critical epitopes on one or more antigen proteins of synthetic;

2) said one or more saltant epi-positions are with one or appear at multiple form on the polypeptide of synthetic;

3) said epitope polypeptide utilizes the MAP technological system directly to synthesize on the dendrimer;

4) dendrimer of said epitope polypeptide is equipped with that proper adjuvant is prepared into dissociation-resisting epitope vaccine or resistance is different-multiepitope epitope vaccine.