Multivalent streptococcus pneumoniae capsular polysaccharide-protein conjugate composition, preparation method therefor, and use thereof

Abstract

Disclosed are a multivalent Streptococcus pneumoniae capsular polysaccharide-protein conjugate composition and a preparation method therefor. The Streptococcus pneumoniae serotype is selected from 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19F, 19A, 20, 22F, 23F, and 33F. The composition can effectively stimulate the production of helper T cells in the body, promote the activation of B cells, and result in immune memory, so as to enable the antibody class to switch from IgM to IgG and enhance immunogenicity, thereby enabling the infants under 2 years of age to produce effective immune responses against corresponding diseases.

Description

A multivalent Streptococcus pneumoniae capsular polysaccharide-protein conjugate composition and its preparation method and application Technical Field

[0001] The present invention relates to a multivalent Streptococcus pneumoniae capsular polysaccharide-protein conjugate composition, a preparation method and an application thereof, and particularly to a 24-valent Streptococcus pneumoniae capsular polysaccharide-protein conjugate composition. Background Art

[0002] Pneumococcal disease (PD) is a serious global public health problem. Pneumococcus (Streptococcus pneumoniae, Spn) is the main pathogen causing serious diseases such as pneumonia, meningitis, and bacteremia in children. It is also a common cause of acute otitis media and sinusitis.

[0003] Currently available vaccines include the following: Pneumococcal vaccines include pneumococcal polysaccharide vaccine (PPV) and pneumococcal conjugate vaccine (PCV). The former primarily includes PPV23, while the latter primarily includes PCV7, PCV10, PCV13, PCV15, and PCV20.

[0004] Pneumococcal conjugate vaccines are currently the mainstream products on the market, including Pfizer, GlaxoSmithKline, etc., in which protein carriers are used in two ways: single and mixed carriers. For example, Pfizer's PCV7 vaccine (PREVNAR) contains capsular polysaccharides from the seven most prevalent serotypes: 4, 6B, 9V, 14, 18C, 19F and 23F. Different capsular polysaccharides and CRMs 197 PCV13 vaccine also uses a single carrier, CRM 197 In mixed vectors, two or more protein carriers are used, with certain specific serotypes being conjugated to specific protein carriers. For example, GlaxoSmithKline's SYNFLORIX conjugates serotypes 1, 4, 5, 6B, 7F, 9V, 14, and 23F to Haemophilus influenzae (H. influenzae) protein D, serotype 18C to tetanus toxoid, and serotype 19F to diphtheria toxoid.

[0005] With the launch and promotion of the above-mentioned 13-valent products, the number of diseases caused by the serotypes included in the existing 13-valent vaccines has generally decreased, while other non-vaccine serotypes, which were previously relatively less common, have gradually received attention. Therefore, there is an urgent need to develop a new pneumonia vaccine to cope with environmental changes. In the development of new vaccines, it is faced with the challenge of adding more serotypes to improve protection. At the same time, it is necessary to consider the selection preferences of different serotype capsular polysaccharides for carrier proteins, and consider the impact of combining multiple polysaccharide-protein conjugates on the immune effects of different serotypes of pneumococci. Based on this, the present invention conducts further research on the above situation.

[0006] Summary of the Invention

[0007] The present invention provides a multivalent Streptococcus pneumoniae capsular polysaccharide-protein conjugate composition, a preparation method and an application thereof.

[0008] Specifically, the present application provides a multivalent Streptococcus pneumoniae capsular polysaccharide-protein conjugate composition, which comprises 24 different Streptococcus pneumoniae capsular polysaccharide-carrier protein conjugates, wherein the Streptococcus pneumoniae serotype is selected from 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19F, 19A, 20, 22F, 23F and 33F, and the carrier protein comprises a first carrier protein and a second carrier protein, wherein the first carrier protein and the second carrier protein are different, and the serotype 12F can be conjugated to the first carrier protein or the second carrier protein.

[0009] Specifically, the pneumococcal capsular polysaccharide serotypes 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 9N, 10A, 11A, 14, 18C, 19A, 20, 22F, 23F and 33F are conjugated to a first carrier protein, and 7F, 15B, 17F, 19F are conjugated to a second carrier protein; or, the pneumococcal capsular polysaccharide serotypes 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 10A, 11A, 14, 15B, 17F, 18C, 19A, 20, 22F, 23F and 33F are conjugated to a first carrier protein, and the pneumococcal capsular polysaccharide 7F, 9N, 19F is conjugated to a second carrier protein; or, the pneumococcal capsular polysaccharide serotypes 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 10A, 11A, 14, 17F, 18C, 19A, 20, 22F, 23F and 33F are conjugated to a first carrier protein, and the pneumococcal capsular polysaccharide 7F, 9N, 15B, 19F is conjugated to a second carrier protein; or, the pneumococcal capsular polysaccharide serotypes 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 10A, 11A, 14, 15B, 18C, 19A, 20, 22F, 23F and 33F are conjugated to a first carrier protein, and the pneumococcal capsular polysaccharide 7F, 9N, 17F, 19F is conjugated to a second carrier protein; or, the pneumococcal capsular polysaccharide serotypes 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 10A, 11A, 14, 18C, 19A, 20, 22F, 23F and 33F are conjugated to a first carrier protein, and the pneumococcal capsular polysaccharide 7F, 9N, 15B, 17F, 19F is conjugated to a second carrier protein. The serotype 12F can be conjugated to the first carrier protein or the second carrier protein.

[0010] Preferably, the pneumococcal capsular polysaccharide 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 9N, 10A, 11A, 14, 18C, 19A, 20, 22F, 23F and 33F are conjugated to a first carrier protein, and the pneumococcal capsular polysaccharide 7F, 12F, 15B, 17F, 19F are conjugated to a second carrier protein.

[0011] Preferably, the pneumococcal capsular polysaccharide 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 10A, 11A, 14, 15B, 17F, 18C, 19A, 20, 22F, 23F and 33F are conjugated to a first carrier protein, and the pneumococcal capsular polysaccharide 7F, 9N, 12F, 19F are conjugated to a second carrier protein.

[0012] Preferably, the pneumococcal capsular polysaccharide 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 10A, 11A, 12F, 14, 17F, 18C, 19A, 20, 22F, 23F and 33F are conjugated to a first carrier protein, and the pneumococcal capsular polysaccharide 7F, 9N, 15B, 19F are conjugated to a second carrier protein.

[0013] Preferably, the pneumococcal capsular polysaccharide 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 20, 22F, 23F and 33F are conjugated to a first carrier protein, and the pneumococcal capsular polysaccharide 7F, 9N, 17F, 19F are conjugated to a second carrier protein.

[0014] Preferably, the pneumococcal capsular polysaccharide 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 10A, 11A, 12F, 14, 18C, 19A, 20, 22F, 23F and 33F are conjugated to a first carrier protein, and the pneumococcal capsular polysaccharide 7F, 9N, 15B, 17F, 19F are conjugated to a second carrier protein.

[0015] Specifically, the carrier protein should be non-toxic, capable of binding to the pneumococcal capsular polysaccharide and causing a significant increase in the immunogenicity of the polysaccharide, and should be easy to obtain in large quantities.

[0016] Specifically, the first carrier protein or the second carrier protein is selected from: CRM 197 , TT (Tetanus toxoid), PD (Haemophilus influenzae protein D), pertussis toxoid (PT), fHbp.

[0017] Specifically, the first carrier protein is: CRM 197 , the second carrier protein is TT or PD or PT.

[0018] Preferably, the first carrier protein is: CRM 197 , the second carrier protein is TT.

[0019] Specifically, the carrier protein of the present invention is obtained through steps such as bacterial fermentation, ultrafiltration, purification, and detoxification.

[0020] CRM of the present invention 197 Non-toxigenic phage β197 tox- Produced by Corynebacterium diphtheriae infection (created by nitrosoguanidine mutagenesis of toxigenic β-rostral phage). CRM 197The protein has the same molecular weight as diphtheria toxin but differs from it by a single base change in the structural gene (guanine to adenine). This single base change causes an amino acid substitution in the mature protein (glutamic acid instead of glycine) and eliminates the toxicity of diphtheria toxin.

[0021] TT (Tetanus toxoid) and DT (diphtheria toxoid) of the present invention, the molecular weights of TT and DT are 151 kilodaltons (kD) and 58kD respectively, which are exotoxins produced by Bacillus tetani and Corynebacterium diphtheriae. They are detoxified with formaldehyde to lose their toxicity and then purified by chemical methods to remove invalid protein impurities, thereby obtaining highly purified toxins. They have been used for many years as vaccines for the prevention of tetanus and diphtheria, and have been selected as carrier molecules at the beginning of the research and development of polysaccharide-protein conjugate vaccines due to their good immunogenicity and safety. Toxoids as carrier proteins also have certain limitations, such as being more stringent in the requirements for Bacillus tetani culture conditions in vaccine production, requiring the processing of a large amount of toxin supernatants during preparation, and the detoxified toxins, in addition to the possibility of residual toxicity and toxic reversal, also undergo certain conformational changes or modifications in their protein structure, which may destroy some T cell antigen sites, thereby affecting immunogenicity.

[0022] The PD described in the present invention is a surface lipoprotein with a relative molecular mass of 42,000, which exists in all Hi strains and is highly conserved. 。

[0023] Specifically, the present invention also provides a method for preparing the multivalent Streptococcus pneumoniae capsular polysaccharide-protein conjugate composition, which comprises separating and purifying the capsular polysaccharides of 24 different serotypes (1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F) of Streptococcus pneumoniae, and then covalently linking the capsular polysaccharides of each serotype to a carrier protein to form a monovalent conjugate. Finally, the monovalent conjugates of each type are mixed to obtain a multivalent Streptococcus pneumoniae capsular polysaccharide-protein conjugate composition.

[0024] Specifically, the steps of separating and purifying Streptococcus pneumoniae capsular polysaccharide are as follows:

[0025] A pneumococcal strain library was established, and pneumococcal fermentation was performed using working seeds. An appropriate amount of sodium deoxycholate was added to the pneumococcal fermentation broth for lysis. The lysed fermentation broth was centrifuged or filtered to remove the bacteria, and the supernatant or filtrate was collected and then concentrated by ultrafiltration. The fermentation recovery obtained by centrifugation or filtration was purified by acid precipitation or alcohol precipitation and column chromatography to remove proteins and cell debris that had not yet precipitated during centrifugation. The filtrate was concentrated on a 100 kDa membrane package and dialyzed to obtain a sample. After ultrafiltration, the ultrafiltration recovery was filtered through a 0.2 μm filter. The filtrate was subjected to in-process quality control (appearance, protein, nucleic acid content, endotoxin, molecular weight, and total amount of polysaccharides). The purified polysaccharide was sterile filtered and stored at -20°C.

[0026] Specifically, the polysaccharide of the present invention can also be purified by other purification techniques well known in the art, such as column chromatography, etc. to purify the capsular polysaccharide of Streptococcus pneumoniae.

[0027] Specifically, the composition of the present invention further comprises an optional excipient and / or an aluminum-based adjuvant.

[0028] The aluminum-based adjuvants include aluminum oxide, aluminum hydroxide, aluminum phosphate, aluminum sulfate, etc. The aluminum-based adjuvants are commercially available and can also be prepared using conventional methods in the art. The excipients are preferably sodium chloride and / or succinic acid.

[0029] The present invention also provides use of the multivalent Streptococcus pneumoniae capsular polysaccharide-protein conjugate composition in the preparation of a medicament for preventing or treating diseases caused by Streptococcus pneumoniae.

[0030] Specifically, the polyvalent pneumococcal polysaccharide-protein conjugate composition can be in the form of a spray, injection, lyophilized agent, capsule, tablet or pill, etc. Immunization can be performed by the following methods: skin, injection (including subcutaneous injection, intramuscular injection, intravenous injection), oral administration, nasal drops or mucosal spray, etc.

[0031] Specifically, the preparation method of the multivalent pneumococcal polysaccharide-protein conjugate composition includes: adsorbing each type of monovalent conjugate with an aluminum adjuvant and then mixing; or mixing each type of monovalent conjugate and then adsorbing with an aluminum adjuvant to obtain a 24-valent pneumococcal capsular polysaccharide conjugate vaccine.

[0032] The multivalent pneumococcal capsular polysaccharide conjugate vaccine of the present invention can be used to protect or treat patients with pneumonia caused by pneumococcal infection. The vaccine can be injected intramuscularly, subcutaneously, intradermally or administered through the mucosa and is suitable for vaccination of humans and animals.

[0033] The beneficial effects of the present invention include:

[0034] In the first aspect, the present invention is aimed at the prevalence of serotypes of Streptococcus pneumoniae unique to my country, and creatively selects serotype combinations of 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F, thereby developing a multivalent Streptococcus pneumoniae capsular polysaccharide-protein conjugate composition designed for the prevalence of serotypes of Streptococcus pneumoniae unique to my country.

[0035] Secondly, the present invention further studies 24 different serotypes, conducts immunogenicity studies on different serotypes and carrier protein combinations, and simultaneously studies the immunogenicity of 24 polysaccharide-carrier protein compositions to screen out the best combination of polysaccharide and carrier protein.

[0036] Thirdly, the present invention further studies 24 different serotypes and rationally controls the content of carrier protein while ensuring maximum immunogenicity.

[0037] Fourthly, the composition of the present invention has multiple immunogenicities and protective properties against the invasion of the above-mentioned 24 serotypes of Streptococcus pneumoniae, which is better than the low-priced pneumonia composition already on the market, and the immune response is higher than that of the uncombined composition.

[0038] Fifthly, the immunogenicity of the multivalent pneumococcal capsular polysaccharide conjugate vaccine of the present invention is much higher than that of similar multivalent polysaccharide vaccines. The composition of the present invention can effectively stimulate the body to produce helper T cells, promote B cell activation and produce immune memory, enable the antibody class to be converted from IgM to IgG, improve immunogenicity, and enable infants and young children under 2 years old to produce effective immune responses against corresponding diseases.

[0039] In a sixth aspect, the multivalent pneumococcal capsular polysaccharide conjugate vaccine of the present invention can prevent invasive diseases caused by the above 24 serotypes of pneumococcus, and is more convenient, multi-effective and has lower cost.

[0040] Seventh, the polyvalent pneumococcal capsular polysaccharide-protein conjugate composition exhibits complete adsorption, stable physicochemical properties, and no mutual interference. Animal experiments have demonstrated that the polyvalent pneumococcal capsular polysaccharide conjugate vaccine of the present invention can effectively induce the production of high-titer, specific antibodies against the aforementioned 24 serotype capsular polysaccharides. In vitro experiments have shown that the antibodies induced by the polyvalent conjugate vaccine exhibit significant reactivity and neutralizing activity. In particular, the immune effect of each serotype capsular polysaccharide in the composition is unaffected by the other components. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1: Immunogenicity of conjugates of type 2 capsular polysaccharide conjugated to different carrier proteins;

[0042] Figure 2: Immunogenicity of conjugates of type 8 capsular polysaccharide conjugated to different carrier proteins;

[0043] Figure 3: Immunogenicity of conjugates of 9N capsular polysaccharide conjugated to different carrier proteins;

[0044] Figure 4: Immunogenicity of conjugates of type 10A capsular polysaccharide conjugated to different carrier proteins;

[0045] Figure 5: Immunogenicity of conjugates of type 11A capsular polysaccharide conjugated to different carrier proteins;

[0046] Figure 6: Immunogenicity of conjugates of 12F capsular polysaccharide conjugated to different carrier proteins;

[0047] Figure 7: Immunogenicity of conjugates of type 15B capsular polysaccharide conjugated to different carrier proteins;

[0048] Figure 8: Immunogenicity of conjugates of 17F capsular polysaccharide conjugated to different carrier proteins;

[0049] Figure 9: Immunogenicity of conjugates of type 20 capsular polysaccharide conjugated to different carrier proteins;

[0050] Figure 10: Immunogenicity of conjugates obtained by combining 22F capsular polysaccharide with different carrier proteins;

[0051] Figure 11: Immunogenicity of conjugates of 33F capsular polysaccharide conjugated to different carrier proteins;

[0052] Figure 12: Antibody titers of 24-valent pneumococcal capsular polysaccharide carrier protein conjugate vaccine against serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F;

[0053] Figure 13: Antibody titers of 24-valent pneumococcal capsular polysaccharide carrier protein conjugate vaccine against serotypes 2, 8, 9N, 10A, 11A, 12F, 15B, 17F, 20, 22F, and 33F, respectively. DETAILED DESCRIPTION

[0054] The following will clearly and completely describe the technical solutions of the present invention in conjunction with the accompanying drawings. Obviously, the embodiments described are only some of the embodiments of the present invention, not all of them. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making any creative efforts shall fall within the scope of protection of the present invention.

[0055] Example 1: Preparation of Streptococcus pneumoniae capsular polysaccharide stock solution

[0056] Serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F, and 33F of Streptococcus pneumoniae were obtained from the National Center for Medical Culture Collections (CMCC). These strains were passaged to establish an original seed bank, a master seed bank, and a working seed bank. The working seed batches were opened and cultured in fermentation tanks, and each generation of seeds was freeze-dried and preserved. The working seeds were streaked on the culture medium to select the strain, inoculated into 5 ml of liquid comprehensive culture medium, cultured at 35°C ± 2°C, and then transferred to 500 ml of culture fluid. The culture was then expanded to 10 L of culture fluid and 50 L of culture fluid. The culture is terminated at the end of the logarithmic growth phase or the beginning of the stationary phase, and samples are taken for bacterial purity testing. After passing the purity test, an appropriate amount of sterile sodium deoxycholate is added to the harvested culture medium to lyse the bacterial cells and release the capsular polysaccharide. The sterilized culture medium is centrifuged, and the supernatant is collected and concentrated by ultrafiltration. The supernatant is further ultrafiltered, exchanged, and purified to obtain 24 purified pneumococcal capsular polysaccharides of serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F, and 33F.

[0057] Example 2: Preparation of 24 Capsular Polysaccharide-Protein Conjugate Compositions and Immunogenicity Testing

[0058] The 24 pneumococcal capsular polysaccharide stock solutions prepared in Example 1 were conjugated with different proteins.

[0059] Capsular polysaccharides of pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 9V, 14, 18C, 19A and 23F were purified by acetic acid hydrolysis, hydrogen peroxide hydrolysis or high pressure degradation to reduce the molecular weight of the polysaccharide, and then hydrolyzed / degraded by sodium periodate or CDAP activation and combined with CRM197.

[0060] The capsular polysaccharides of serotypes 7F and 19F pneumococci were hydrolyzed with acetic acid or hydrogen peroxide or degraded under high pressure to reduce the molecular weight of the purified polysaccharides. The polysaccharides were then hydrolyzed / degraded by CDAP activation and bound to TT.

[0061] Capsular polysaccharides of pneumococcal serotypes 2, 8, 9N, 10A, 11A, 12F, 15B, 17F, 20, 22F and 33F were hydrolyzed with acetic acid or hydrogen peroxide or degraded under high pressure to reduce the molecular weight of the purified polysaccharides. The polysaccharides were then activated and hydrolyzed / degraded by sodium periodate or CDAP and combined with CRM197, TT and PD protein carriers, respectively.

[0062] The polysaccharide-protein conjugate has been subjected to quality analysis, and the free polysaccharide content is controlled within 30%, the free protein content is within 5%, the polysaccharide-protein ratio is between 0.3-2.5, and the endotoxin and other impurity contents are controlled within a safe and acceptable range.

[0063] The immunogenicity of pneumococcal capsular polysaccharides of serotypes 2, 8, 9N, 10A, 11A, 12F, 15B, 17F, 20, 22F, and 33F combined with CRM197, TT, and PD carrier proteins was tested. The results are shown in Figures 1 to 11.

[0064] The results showed that for serotypes 2, 8, 10A and 11A, the immunogenicity was stronger when combined with the CRM197 carrier protein than when combined with the TT carrier protein.

[0065] Example 3: Preparation of a polyvalent Streptococcus pneumoniae capsular polysaccharide-protein conjugate composition and its immunogenicity detection

[0066] Based on the previous 13-valent pneumococcal combination research composition, 1, 3, 4, 5, 6A, 6B, 9V, 14, 18C, 19A, and 23F were selected to bind to CRM197, and 7F and 19F to bind to TT. Other serotypes were further screened for specific binding carrier proteins in the composition based on the experimental results of Example 2, as shown in Table 1 below (the CRM mentioned in the table specifically refers to CRM197).

[0067] Table 1. Screening of new serum type carrier proteins in polysaccharide-protein conjugate compositions

[0068] The prepared polysaccharide-protein monovalent conjugates 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F, and 33F are diluted with a buffer solution having a pH of 5.3 to 6.8, mixed, and then added with an aluminum phosphate adjuvant and stirred to prepare a semi-finished product. The polysaccharide content of each conjugate is 2 to 8 μg per milliliter, and the aluminum phosphate adjuvant content is 0.125 to 0.5 mg per milliliter.

[0069] Immunization protocol: The polyvalent vaccine stock solution was added to an aluminum phosphate adjuvant to prepare an immune antigen. Four New Zealand white rabbits, weighing 2.0-2.5 kg, were immunized intramuscularly in the thigh. Each immunization dose was 0.25 ml. Blood was drawn 35 days later to assess the immunogenicity of the polysaccharide. The immunization results are shown in Figures 12 and 13.

[0070] The results showed that different carrier protein combinations resulted in different immune responses. Groups 2-7, PCV24 polysaccharide conjugate vaccine compositions containing some serotypes conjugated to the TT carrier protein, demonstrated superior immune efficacy compared to Group 1, where the entire carrier protein was CRM197. Overall immune efficacy was higher when serotypes 1, 2, 3, 4, 5, 6A, 6B, 8, 9N, 9V, 10A, 11A, 14, 18C, 19A, 20, 22F, 23F, and 33F were conjugated to CRM197; 7F, 15B, 17F, and 19F were conjugated to TT; and 12F was conjugated to either CRM197 or TT.

[0071] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

[0072] The aforementioned embodiments and methods described in the present invention may vary based on the abilities, experiences and preferences of those skilled in the art. The present invention does not limit the order of the method steps by simply listing the steps in a certain order.

Claims

1. A multivalent pneumococcal capsular polysaccharide-protein conjugate composition, comprising pneumococcal capsular polysaccharides from 24 different serotypes and a carrier protein, characterized in that: The pneumococcal capsular polysaccharides from different serotypes are selected from 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19F, 19A, 20, 22F, 23F and 33F, and the carrier protein includes a first carrier protein and a second carrier protein, and the first carrier protein and the second carrier protein are different; The serotype 12F polysaccharide is conjugated to the first carrier protein or the second carrier protein, the serotypes 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 9N, 10A, 11A, 14, 18C, 19A, 20, 22F, 23F and 33F polysaccharides are conjugated to the first carrier protein, and the serotypes 7F, 15B, 17F, 19F polysaccharides are conjugated to the second carrier protein; or, the serotype 12F polysaccharide is conjugated to the first carrier protein or the second carrier protein, and the serotype 1 , 2, 3, 4, 5, 6A, 6B, 8, 9V, 10A, 11A, 14, 15B, 17F, 18C, 19A, 20, 22F, 23F and 33F polysaccharides are conjugated to the first carrier protein, and the polysaccharides of serotypes 7F, 9N, 19F are conjugated to the second carrier protein; or, the polysaccharide of serotype 12F is conjugated to the first carrier protein or the second carrier protein, and the polysaccharides of serotypes 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 10A, 11A, 14, 17 The polysaccharide of serotype F, 18C, 19A, 20, 22F, 23F and 33F is conjugated to the first carrier protein, and the polysaccharide of serotype 7F, 9N, 15B, 19F is conjugated to the second carrier protein; or, the polysaccharide of serotype 12F is conjugated to the first carrier protein or the second carrier protein, and the polysaccharide of serotype 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 10A, 11A, 14, 15B, 18C, 19A, 20, 22F, 23F and 33F is conjugated to the first carrier protein. protein, the polysaccharide of serotype 7F, 9N, 17F, 19F is conjugated to a second carrier protein; or, the polysaccharide of serotype 12F is conjugated to a first carrier protein or a second carrier protein, the polysaccharide of serotypes 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 10A, 11A, 14, 18C, 19A, 20, 22F, 23F and 33F is conjugated to a first carrier protein, and the polysaccharide of serotypes 7F, 9N, 15B, 17F, 19F is conjugated to a second carrier protein.

2. The composition according to claim 1, characterized in that The polysaccharides of serotypes 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 9N, 10A, 11A, 14, 18C, 19A, 20, 22F, 23F and 33F are conjugated to a first carrier protein, and the polysaccharides of serotypes 7F, 12F, 15B, 17F and 19F are conjugated to a second carrier protein; or, the polysaccharide of serotype 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 10A, 11A, 14, 15B, 17F, 18C, 19A, 20, 22F, 23F and 33F is conjugated to a first carrier protein, and the polysaccharide of serotype 7F, 9N, 12F, 19F is conjugated to a second carrier protein; or, the polysaccharide of serotype 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 10A, 11A, 12F, 14, 17F, 18C, 19A, 20, 22F, 23F and 33F is conjugated to a first carrier protein, and the polysaccharide of serotype 7F, 9N, 15B, 19F is conjugated to a second carrier protein; or, the polysaccharide of serotype 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19A, 20, 22F, 23F and 33F is conjugated to a first carrier protein, and the polysaccharide of serotype 7F, 9N, 17F, 19F is conjugated to a second carrier protein; Or, the polysaccharides of serotypes 1, 2, 3, 4, 5, 6A, 6B, 8, 9V, 10A, 11A, 12F, 14, 18C, 19A, 20, 22F, 23F and 33F are conjugated to a first carrier protein, and the polysaccharides of serotypes 7F, 9N, 15B, 17F, 19F are conjugated to a second carrier protein.

3. The composition according to claim 1 or 2, characterized in that The first carrier protein and the second carrier protein are selected from: CRM197, TT, PD, PT, fHbp.

4. The composition according to claim 3, characterized in that The first carrier protein is CRM197, and the second carrier protein is TT, PD or PT; preferably, the first carrier protein is CRM197, and the second carrier protein is TT.

5. The composition according to any one of claims 1 to 4, characterized in that The composition further comprises an excipient and / or an aluminum-based adjuvant.

6. The composition according to any one of claims 5, characterized in that The aluminum-based adjuvant is selected from the group consisting of aluminum oxide, aluminum hydroxide, aluminum phosphate and aluminum sulfate.

7. The composition according to any one of claims 1 to 6, characterized in that The composition is prepared as spray, injection, lyophilized preparation, capsule, tablet or pill.

8. The composition according to any one of claims 1 to 7, characterized in that The composition can be immunized by the following methods: skin, injection, oral administration, nasal drops or mucosal spray; preferably, the injection is subcutaneous injection, intramuscular injection or intravenous injection.

9. A method for preparing the multivalent Streptococcus pneumoniae capsular polysaccharide-protein conjugate composition according to any one of claims 1 to 8, characterized in that: The method comprises: respectively adsorbing various types of monovalent conjugates with aluminum adjuvants and then mixing them, or mixing various types of monovalent conjugates and then adsorbing them with aluminum adjuvants to prepare a 24-valent pneumococcal capsular polysaccharide conjugate vaccine.

10. Use of any one of the multivalent pneumococcal capsular polysaccharide-protein conjugate compositions according to claims 1 to 8 in the preparation of vaccines or drugs for preventing or treating diseases, characterized in that: The disease is a disease caused by infection with Streptococcus pneumoniae.

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