K64 type klebsiella pneumoniae capsular polysaccharide monoclonal antibody, and hybridoma cell strain and application thereof

Abstract

The invention discloses a K64 type klebsiella pneumoniae capsular polysaccharide monoclonal antibody, a hybridoma cell strain and application thereof, the sequence information of the klebsiella pneumoniae capsular polysaccharide monoclonal antibody is as shown in SEQ NO: 1-SEQ NO: 56, the preservation numbers of the hybridoma cell strain are respectively CCTCC: C2023365, CCTCC: C2023366, CCTCC: C2023367, CCTCC: C2024340, CCTCC: C2024341, CCTCC: C2024342 and CCTCC: C2024343, and the preservation numbers of the hybridoma cell strain are as shown in SEQ NO: 1-SEQ NO: 56. The klebsiella pneumoniae capsular polysaccharide monoclonal antibody provided by the invention can be specifically combined with capsular polysaccharide of klebsiella pneumoniae, is beneficial to typing of the klebsiella pneumoniae in clinic, and can be proved to have an excellent effect on preventing and treating infection of K64 type klebsiella pneumoniae in animal in-vivo and in-vitro tests; the invention provides a way for solving the problems of klebsiella pneumoniae infection and multiple drug resistance, has obvious significance for preventing, diagnosing and treating klebsiella pneumoniae infection, and has important value for developing a new generation of klebsiella pneumoniae medicine.

Description

Technical Field

[0001] The present application belongs to the field of medical technology, and in particular relates to a monoclonal antibody against K64 type Klebsiella pneumoniae capsular polysaccharide, a hybridoma cell line thereof, and applications thereof. Background Art

[0002] Klebsiella pneumoniae is a Gram-negative bacterium (G-) with a capsule. Klebsiella pneumoniae mainly colonizes the digestive tract and respiratory tract. When the body's resistance is reduced, it can cause lung infection, urinary tract infection, and even bloodstream infection. It is also one of the most common bacteria causing respiratory tract infections.

[0003] Infections caused by Klebsiella pneumoniae have become one of the most serious public health problems in the world. According to statistics, the mortality rate of Klebsiella pneumoniae infection is extremely high. The mortality rate of bloodstream infection caused by Klebsiella pneumoniae is 20% to 30%, while the mortality rate of Klebsiella pneumoniae bacteremia combined with pneumonia can reach more than 50%. The treatment of Klebsiella pneumoniae mainly relies on antibiotics, but in recent years, due to the widespread use of various antimicrobial drugs, Klebsiella pneumoniae that produces extended-spectrum β-lactamases and carbapenemases are resistant to almost all available β-lactams (including carbapenems). In the past 10 years, the rate of carbapenem-resistant Klebsiella pneumoniae (CRKP) has increased dramatically worldwide. Capsular polysaccharides are the main virulence factors of Klebsiella pneumoniae and are important protective antigens for the treatment or prevention of Klebsiella pneumoniae infections. According to the different structures of capsular polysaccharides, Klebsiella pneumoniae can be divided into at least 77 serotypes, which are also called K antigens or K types. Different K types show different resistance profiles, have different distributions in geographical and clinical settings, and may have different epidemic patterns in hospital infections, thus directly affecting the choice of clinical treatment options. Among them, K64 type Klebsiella pneumoniae, although less reported, may still cause serious infections, especially in immunocompromised individuals. Compared with other serotypes, K64 type may show unique pathogenic properties in some cases. Compared with other types, K64 type may have a different antibiotic resistance spectrum and may also have special pathogenic manifestations in certain special environments or populations. However, there are few clinical studies on K64 type Klebsiella pneumoniae.

[0004] Therefore, there is a need in the art to develop a K64-type Klebsiella pneumoniae capsular polysaccharide monoclonal antibody with high specificity, good affinity and strong binding ability, which is beneficial to the prevention and control of drug-resistant Klebsiella pneumoniae and its clinical application. Summary of the invention

[0005] To solve the above technical problems, the present application provides a Klebsiella pneumoniae capsular polysaccharide monoclonal antibody, wherein the Klebsiella pneumoniae is K64 type Klebsiella pneumoniae, and the sequence of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody comprises a light chain variable region and a heavy chain variable region, the amino acid sequence of the light chain variable region is as shown in SEQ NO:1 or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto, and the amino acid sequence of the heavy chain variable region is as shown in SEQ NO:2 or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto.

[0006] Specifically, the complementary determining region of the light chain variable region comprises three sequences as shown in SEQ ID:3, SEQ ID:4 and SEQ ID:5, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:3, SEQ ID:4 and SEQ ID:5; the complementary determining region of the heavy chain variable region comprises three sequences as shown in SEQ ID:6, SEQ ID:7 and SEQ ID:8, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:6, SEQ ID:7 and SEQ ID:8.

[0007] In a second aspect, the present application provides a hybridoma cell strain, the hybridoma cell strain is named K64-1, the deposit number of the hybridoma cell strain is: CCTCC NO: C2023365, and the hybridoma cell strain is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody as described in the first aspect.

[0008] In a third aspect, the present application provides a Klebsiella pneumoniae capsular polysaccharide monoclonal antibody, wherein the Klebsiella pneumoniae is K64 type Klebsiella pneumoniae, and the sequence of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody comprises a light chain variable region and a heavy chain variable region, the amino acid sequence of the light chain variable region is as shown in SEQ NO:9 or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto, and the amino acid sequence of the heavy chain variable region is as shown in SEQ NO:10 or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto.

[0009] Specifically, the complementarity determining region of the light chain variable region comprises three sequences as shown in SEQ ID:11, SEQ ID:12 and SEQ ID:13, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:11, SEQ ID:12 and SEQ ID:13, and the complementarity determining region of the heavy chain variable region comprises three sequences as shown in SEQ ID:14, SEQ ID:15 and SEQ ID:16, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:14, SEQ ID:15 and SEQ ID:16.

[0010] In a fourth aspect, the present application provides a hybridoma cell strain, the hybridoma cell strain is named K64-23, the deposit number of the hybridoma cell strain is: CCTCC NO: C2023366, and the hybridoma cell strain is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody as described in the third aspect.

[0011] In a fifth aspect, the present application provides a Klebsiella pneumoniae capsular polysaccharide monoclonal antibody, wherein the Klebsiella pneumoniae is K64 type Klebsiella pneumoniae, and the sequence of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody comprises a light chain variable region and a heavy chain variable region, the amino acid sequence of the light chain variable region is as shown in SEQ NO: 17 or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto, and the amino acid sequence of the heavy chain variable region is as shown in SEQ NO: 18 or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto.

[0012] Specifically, the complementarity determining region of the light chain variable region comprises three sequences as shown in SEQ ID: 19, SEQ ID: 20 and SEQ ID: 21, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID: 19, SEQ ID: 20 and SEQ ID: 21, and the complementarity determining region of the heavy chain variable region comprises three sequences as shown in SEQ ID: 22, SEQ ID: 23 and SEQ ID: 24, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID: 22, SEQ ID: 23 and SEQ ID: 24.

[0013] In a sixth aspect, the present application provides a hybridoma cell strain, which is named K64-24, and the deposit number of the hybridoma cell strain is: CCTCC NO: C2023367. The hybridoma cell strain is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody as described in the fifth aspect.

[0014] In a seventh aspect, the present application provides a Klebsiella pneumoniae capsular polysaccharide monoclonal antibody, wherein the Klebsiella pneumoniae is K64 type Klebsiella pneumoniae, and the sequence of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody comprises a light chain variable region and a heavy chain variable region, the amino acid sequence of the light chain variable region is as shown in SEQ NO:25, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto, and the amino acid sequence of the heavy chain variable region is as shown in SEQ NO:26, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto.

[0015] Specifically, the complementarity determining region of the light chain variable region comprises three sequences as shown in SEQ ID:27, SEQ ID:28 and SEQ ID:29, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:27, SEQ ID:28 and SEQ ID:29, and the complementarity determining region of the heavy chain variable region comprises three sequences as shown in SEQ ID:30, SEQ ID:31 and SEQ ID:32, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:30, SEQ ID:31 and SEQ ID:32.

[0016] In an eighth aspect, the present application provides a hybridoma cell strain, which is named K64-3, and the deposit number of the hybridoma cell strain is: CCTCC NO: C2024340. The hybridoma cell strain is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody as described in the seventh aspect.

[0017] In a ninth aspect, the present application provides a Klebsiella pneumoniae capsular polysaccharide monoclonal antibody, wherein the Klebsiella pneumoniae is K64 type Klebsiella pneumoniae, and the sequence of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody comprises a light chain variable region and a heavy chain variable region, the amino acid sequence of the light chain variable region is as shown in SEQ NO:33 or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto, and the amino acid sequence of the heavy chain variable region is as shown in SEQ NO:34 or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto.

[0018] Specifically, the complementarity determining region of the light chain variable region comprises three sequences as shown in SEQ ID:35, SEQ ID:36 and SEQ ID:37, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:35, SEQ ID:36 and SEQ ID:37, and the complementarity determining region of the heavy chain variable region comprises three sequences as shown in SEQ ID:38, SEQ ID:39 and SEQ ID:40, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:38, SEQ ID:39 and SEQ ID:40.

[0019] In the tenth aspect, the present application provides a hybridoma cell strain, which is named K64-5, and the deposit number of the hybridoma cell strain is: CCTCC NO: C2024341. The hybridoma cell strain is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody as described in the ninth aspect.

[0020] In the eleventh aspect, the present application provides a Klebsiella pneumoniae capsular polysaccharide monoclonal antibody, wherein the Klebsiella pneumoniae is K64 type Klebsiella pneumoniae, and the sequence of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody comprises a light chain variable region and a heavy chain variable region, the amino acid sequence of the light chain variable region is as shown in SEQ NO:41, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto, and the amino acid sequence of the heavy chain variable region is as shown in SEQ NO:42, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto.

[0021] Specifically, the complementarity determining region of the light chain variable region comprises three sequences as shown in SEQ ID:43, SEQ ID:44 and SEQ ID:45, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:43, SEQ ID:44 and SEQ ID:45, and the complementarity determining region of the heavy chain variable region comprises three sequences as shown in SEQ ID:46, SEQ ID:47 and SEQ ID:48, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:46, SEQ ID:47 and SEQ ID:48.

[0022] In the twelfth aspect, the present application provides a hybridoma cell strain, which is named K64-8 and has a deposit number of CCTCC NO: C2024342. The hybridoma cell strain is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody as described in the eleventh aspect.

[0023] In a thirteenth aspect, the present application provides a Klebsiella pneumoniae capsular polysaccharide monoclonal antibody, wherein the Klebsiella pneumoniae is K64 type Klebsiella pneumoniae, and the sequence of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody comprises a light chain variable region and a heavy chain variable region, the amino acid sequence of the light chain variable region is as shown in SEQ NO:49 or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto, and the amino acid sequence of the heavy chain variable region is as shown in SEQ NO:50 or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto.

[0024] Specifically, the complementarity determining region of the light chain variable region comprises three sequences as shown in SEQ ID:51, SEQ ID:52 and SEQ ID:53, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:51, SEQ ID:52 and SEQ ID:53; the complementarity determining region of the heavy chain variable region comprises three sequences as shown in SEQ ID:54, SEQ ID:55 and SEQ ID:56, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:54, SEQ ID:55 and SEQ ID:56.

[0025] In the fourteenth aspect, the present application provides a hybridoma cell strain, the hybridoma cell strain is named K64-27, the deposit number of the hybridoma cell strain is: CCTCC NO: C2024343, and the hybridoma cell strain is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody as described in the fifteenth aspect.

[0026] In a fifteenth aspect, the present application provides a pharmaceutical composition, comprising the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody described in any of the foregoing, wherein the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody can bind to the Klebsiella pneumoniae capsular polysaccharide target antigen and the non-Klebsiella pneumoniae capsular polysaccharide target antigen, and the pharmaceutical composition further comprises any one or more combinations of pharmaceutically acceptable carriers, excipients or diluents.

[0027] In the sixteenth aspect, the present application provides a kit for detecting K64 type Klebsiella pneumoniae, wherein the kit for detecting K64 type Klebsiella pneumoniae comprises the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody described in any of the foregoing, and the kit for detecting K64 type Klebsiella pneumoniae also comprises a positive control, a negative control, an antibody diluent, a color developing solution, a stopping solution, a blocking solution or a washing solution, any one or more combinations thereof.

[0028] In the seventeenth aspect, the present application provides the use of any of the aforementioned Klebsiella pneumoniae capsular polysaccharide monoclonal antibodies in the preparation of K64 type Klebsiella pneumoniae infection disease treatment drugs and / or detection products, wherein the disease comprises one or more symptoms caused by Klebsiella infection, including pneumonia, urinary tract infection, sepsis / bacteremia / sepsis, neonatal sepsis I bacteremia / sepsis, diarrhea, soft tissue infection, post-organ transplant infection, surgical infection, wound infection, lung infection, suppurative liver abscess, lung abscess, cellulitis, necrotizing fasciitis, myositis, endophthalmitis, peritonitis, meningitis, necrotizing meningitis or spondyloarthropathy.

[0029] Capsular polysaccharide is an important toxic factor of Klebsiella pneumoniae. The present application uses capsular polysaccharide coupled to carrier protein to improve immunogenicity, and uses hybridoma technology to prepare K64 type Klebsiella pneumoniae capsular polysaccharide-specific monoclonal antibodies, which have the characteristics of good specificity, high affinity, and high binding ability with Klebsiella pneumoniae capsular polysaccharide.

[0030] The beneficial effects of this application are:

[0031] The Klebsiella pneumoniae capsular polysaccharide monoclonal antibody provided in the present application can specifically bind to the capsular polysaccharide of Klebsiella pneumoniae type K64, which is beneficial to the typing of Klebsiella pneumoniae in clinical practice. It can be proved in both in vitro and in vivo animal experiments that it has excellent effects in preventing and treating K64 type Klebsiella pneumoniae infection, provides a way to solve the problems of Klebsiella pneumoniae infection and multidrug resistance, has significant significance for the prevention, diagnosis and treatment of Klebsiella pneumoniae infection, and is of great value for the development of a new generation of drugs for Klebsiella pneumoniae. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 This is a graph showing the titer level of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody provided by the present application detected by indirect ELISA method in Example 2;

[0033] Figure 2 This is a graph showing the binding results of flow cytometry analysis of K64 monoclonal antibody, negative control and K64-type Klebsiella pneumoniae Y8 in Example 4;

[0034] Figure 3The bactericidal curves of monoclonal antibodies K64-1, K64-23 and K64-24 against a highly virulent strain of Klebsiella pneumoniae (numbered Y8) in vitro in Example 5;

[0035] Figure 4 This is the survival curve of the K64-1 antibody preventive trial in Example 6. DETAILED DESCRIPTION

[0036] The technical solution of the present application will be described clearly and completely below. Obviously, the described embodiments are part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in the field without creative work are within the scope of protection of the present application.

[0037] The following is the strain deposit information:

[0038] The hybridoma cell line K64-1 (CCTCC NO: C2023365), hybridoma cell line K64-23 (CCTCC NO: C2023366) and hybridoma cell line K64-24 (CCTCC NO: C2023367) provided in this application have all been deposited in the China Center for Type Culture Collection on November 28, 2023.

[0039] The hybridoma cell line K64-3 (CCTCC NO: C2024340), hybridoma cell line K64-5 (CCTCCNO: C2024341), hybridoma cell line K64-8 (CCTCC NO: C2024342) and hybridoma cell line K64-27 (CCTCCNO: C2024343) provided in the present application have all been deposited in the China Center for Type Culture Collection on September 25, 2024.

[0040] Deposit address: Wuhan University, Wuhan, Hubei Province.

[0041] Deposit name and deposit number:

[0042] Hybridoma cell line K64-1: Hybridoma cell line K64-1, CCTCC NO: C2023365.

[0043] Hybridoma cell line K64-23: Hybridoma cell line K64-23, CCTCC NO: C2023366.

[0044] Hybridoma cell line K64-24: Hybridoma cell line K64-24, CCTCC NO: C2023367.

[0045] Hybridoma cell line K64-3: Hybridoma cell line K64-3, CCTCC NO: C2024340.

[0046] Hybridoma cell line K64-5: Hybridoma cell line K64-5, CCTCC NO: C2024341.

[0047] Hybridoma cell line K64-8: Hybridoma cell line K64-8, CCTCC NO: C2024342.

[0048] Hybridoma cell line K64-27: Hybridoma cell line K64-27, CCTCC NO: C2024343.

[0049] In Examples 4 and 5 of the present application, the light chain variable region (VL) sequence of the negative control antibody used (as shown in SEQ ID NO: 57) is:

[0050] MESHSQVFIFLLFWIPVSRGDILLTQSPAILSVSPGERVSFSCRASQSIGRSIHWYQQRTN

[0051] GSPRLLIKYASESIYGIPSRFSGSGSGTDFTLSINSVESEDIAAYYCQQSYNWPRTFGGGT

[0052] KLEIK

[0053] The negative control antibody heavy chain variable region (VH) sequence used (as shown in SEQ ID NO: 58):

[0054] MNFGLSWVFLVALLNGVQCQVQLVETGGGLVRPGNSLNLSCITSGFTFSNYRLHWLRQ

[0055] PPGKGLEWLAVIAVKSDNFGAIYADSVKGRFTISRDDSRSSVYLQMNRLREEDTATYYC

[0056] VRAGVSFFDYWGQGTSLTVSS

[0057] Example 1: Preparation of monoclonal antibodies against Klebsiella pneumoniae K64 capsular polysaccharide

[0058] 1) Establishment of cell line for monoclonal antibody against Klebsiella pneumoniae K64 capsular polysaccharide

[0059] The K64 type capsular polysaccharide protein conjugate is obtained by chemically coupling the capsular polysaccharide extracted from Klebsiella pneumoniae K64 type (GDMCC No: 62132) after fermentation with a carrier protein (diphtheria toxin non-toxic variant CRM197). The method is a commonly used polysaccharide activation method, such as cyanogen bromide method (reference US6375846B1), 1-cyano-4-dimethylamino arsenic tetrafluoroborate (CDAP) (EP0720485), and periodic acid oxidation method (US4711779). The K64-type polysaccharide protein conjugate was added to Freund's adjuvant and immunized SPF-grade BALB / c mice multiple times. When the immune titer was greater than 1:300,000, the spleen was taken and hybridoma technology was used to obtain three monoclonal cell lines: hybridoma cell line K64-1 (CCTCC NO: C2023365), hybridoma cell line K64-23 (CCTCC NO: C2023366), hybridoma cell line K64-24 (CCTCC NO: C2023367), hybridoma cell line K64-3 (CCTCC NO: C2024340), hybridoma cell line K64-5 (CCTCC NO: C2024341), hybridoma cell line K64-8 (CCTCC NO: C2024342) and hybridoma cell line K64-27 (CCTCC NO: C2024343). NO:C2024343), and its purebred status was verified by sequencing.

[0060] After sequencing, the sequence information of 7 Klebsiella pneumoniae capsular polysaccharide monoclonal antibodies is as follows (SEQ NO: 1 to SEQ NO: 56 in sequence):

[0061] K64-1 light chain variable region (VL) sequence (starting from FR-L1):

[0062] QIVLTQSPAIMSASPGEKVTMTCSASSSISYMHWYQQKPGTSPKRWIYDTSKLASGVPA

[0063] RFSGSGSGTSYSLTISSMEAEDAATYYCHQRSSYPWTFGGGTKLEIK

[0064] K64-1 heavy chain variable region (VH) sequence (starting from FR-H1):

[0065] EVKLVESGGGLVKPGGSLKLSCAGSGFTFSSYGMSWVRQTPDKRLEWVATISGGGSYT

[0066] YYSDSVKGRFTISRDNAKNNLYLQMSSLRSEDTALYYCARNYGYDGHYYAMDYWGQ

[0067] GTSVTVSS

[0068] Complementarity determining region 1 (LCDR1) sequence of K64-1 light chain:

[0069] SASSSISYMH

[0070] Complementarity determining region 2 (LCDR2) sequence of K64-1 light chain:

[0071] DTSKLAS

[0072] Complementarity determining region 3 (LCDR3) sequence of K64-1 light chain:

[0073] HQRSSYPWT

[0074] Complementarity determining region 1 (HCDR1) sequence of K64-1 heavy chain:

[0075] Complementarity determining region 2 (HCDR2) sequence of SYGMS K64-1 heavy chain:

[0076] TISGGGSYTYYSDSVKG K64-1 heavy chain complementarity determining region 3 (HCDR3) sequence:

[0077] NYGYDGHYYAMDY K64-23 light chain variable region (VL) sequence (starting from FR-L1):

[0078] QIVLTQSPAIMSASPGEKVTMTCSATSIVNYMHWYQQKSGTSPKRWIYDTSKLASGVPA

[0079] RFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPRT K64-23 heavy chain variable region (VH) sequence (starting from FR-H1):

[0080] QVQLQQPWSELVRPGASVKLPCKASGYTFTSYWMNWVKQRPEQGLEWIGRIDPYDG

[0081] GTHYSQNFMDRAILTVDRSSNTAYMQLNSLTSEDSAVYYCARGYANGAFVYWGQGTL

[0082] VTVSA

[0083] K64-23 light chain complementarity determining region 1 (LCDR1) sequence:

[0084] Complementarity determining region 2 (LCDR2) sequence of the light chain of SATSIVNYMH K64-23:

[0085] DTSKLAS K64-23 light chain complementarity determining region 3 (LCDR3) sequence:

[0086] QQWSSNPRT K64-23 heavy chain complementarity determining region 1 (HCDR1) sequence:

[0087] Complementarity determining region 2 (HCDR2) sequence of SYWMN K64-23 heavy chain:

[0088] RIDPYDGGTHYSQNFMD K64-23 heavy chain complementarity determining region 3 (HCDR3) sequence:

[0089] GYANGAFVY K64-24 light chain variable region (VL) sequence (starting from FR-L1):

[0090] QIVLTQSPAIMSASPGEKVTISCSASSSVSYMYWYQQRPGSSPKPWIYRTSNLASGVPAR

[0091] FSGSGSGTSYSLTISSMEAEDAATYYCQQYHSYPLTFGAGTKLELK K64-24 heavy chain variable region (VH) sequence (starting from FR-H1):

[0092] EVKLEESGGGLVQPGGSMKLSCAASGFTFSDAYMDWVRQSPEKGLEWVAEIRSKANN

[0093] HATLYAESVKGRFTISRDDSKSSVYLQMNSLRAEDTGIYYCTPYYYGSSQFAYWGQGTL

[0094] VTVSA

[0095] Complementarity determining region 1 (LCDR1) sequence of K64-24 light chain:

[0096] SASSSVSYMY K64-24 light chain complementarity determining region 2 (LCDR2) sequence:

[0097] The complementarity determining region 3 (LCDR3) sequence of the RTSNLAS K64-24 light chain:

[0098] QQYHSYPLT K64-24 heavy chain complementarity determining region 1 (HCDR1) sequence:

[0099] Complementarity determining region 2 (HCDR2) sequence of DAYMD K64-24 heavy chain:

[0100] EIRSKANNHATLYAESVKG K64-24 heavy chain complementarity determining region 3 (HCDR3) sequence:

[0101] YYYGSSQFAY K64-3 light chain variable region (VL) sequence (starting from FR-L1):

[0102] QIVLTQSPAIMSASPGEKVTMTCSASSSISYMHWYQQKPGTSPKRWIYDTSKLASGVPA

[0103] RFSGSGSGTSYSLTISSMEAEDAATYYCHQRSSYPWTFGGGTKLEIK K64-3 heavy chain variable region (VH) sequence (starting from FR-H1):

[0104] EVKLVESGGGLVKPGGSLTLSCAASGSIFSTYGMSWVRQTPEKRLEWVATISGGGRYTY

[0105] YPDSVKGRFTISRDNAKNNLYLQMSSLRSEDTALYYCARNYGYDGHYYPMDYWGQG

[0106] TSVTVSS

[0107] K64-3 light chain complementarity determining region 1 (LCDR1) sequence:

[0108] SASSSISYMH K64-3 light chain complementarity determining region 2 (LCDR2) sequence:

[0109] DTSKLAS K64-3 light chain complementarity determining region 3 (LCDR3) sequence:

[0110] HQRSSYPWT K64-3 heavy chain complementarity determining region 1 (HCDR1) sequence:

[0111] Complementarity determining region 2 (HCDR2) sequence of TYGMS K64-3 heavy chain:

[0112] TISGGGRYTYYPDSVKG K64-3 heavy chain complementarity determining region 3 (HCDR3) sequence:

[0113] NYGYDGHYYPMDY K64-5 light chain variable region (VL) sequence (starting from FR-L1):

[0114] QIVLTQSPAIMSASPGEKVTMTCSASSSISYMHWYQQKPGTSPKRWIYDTSKLASGVPV

[0115] RFSGSGSGTSYSLTISSMEAEDAATYYCHQRSSYPWTFGGGTKLEIK K64-5 heavy chain variable region (VH) sequence (starting from FR-H1):

[0116] EVKLVESGGGLVKPGGSLELSCAASGFTFSTYGMSWVRQTPEKRLEWVATISGGGSYT

[0117] YYVDSVKGRFTISRDNAKNNLYLQMSSLRSEDTALYYCARNYGYDGHYYSMDYWGQ

[0118] GTSVTVSS

[0119] K64-5 light chain complementarity determining region 1 (LCDR1) sequence:

[0120] SASSSISYMH K64-5 light chain complementarity determining region 2 (LCDR2) sequence:

[0121] DTSKLAS K64-5 light chain complementarity determining region 3 (LCDR3) sequence:

[0122] HQRSSYPWT K64-5 heavy chain complementarity determining region 1 (HCDR1) sequence:

[0123] Complementarity determining region 2 (HCDR2) sequence of TYGMS K64-5 heavy chain:

[0124] TISGGGSYTYYVDSVKG K64-5 heavy chain complementarity determining region 3 (HCDR3) sequence:

[0125] NYGYDGHYYSMDY K64-8 light chain variable region (VL) sequence (starting from FR-L1):

[0126] QIVLTQSPAIMSASPGEKVTMTCSASSSISYMHWYQQKPGTSPKRWIYDTSKLASGVPA

[0127] RFSGSGSGTSYSLTISSMEAEDAATYYCHQRSNYPWTFGGGTKLEIK K64-8 heavy chain variable region (VH) sequence (starting from FR-H1):

[0128] EVKLVESGGDLVKPGGSLKLSCAASGFTFSSYGMSWVRQTPEKRLEWVATISGGGYYT

[0129] YYPDSVKGRFTISRDNAKNNLYLQMSSLRSEDTALYYCGRNYGYDGHYYAMDYWGQ

[0130] GTSVTVSS

[0131] Complementarity determining region 1 (LCDR1) sequence of K64-8 light chain:

[0132] SASSSISYMH K64-8 light chain complementarity determining region 2 (LCDR2) sequence:

[0133] DTSKLAS

[0134] Complementarity determining region 3 (LCDR3) sequence of K64-8 light chain:

[0135] HQRSNYPWT

[0136] Complementarity determining region 1 (HCDR1) sequence of K64-8 heavy chain:

[0137] SYGMS

[0138] Complementarity determining region 2 (HCDR2) sequence of K64-8 heavy chain:

[0139] TISGGGYYTYYPDSVKG

[0140] Complementarity determining region 3 (HCDR3) sequence of K64-8 heavy chain:

[0141] NYGYDGHYYAMDY

[0142] K64-27 light chain variable region (VL) sequence (starting from FR-L1):

[0143] QIVLTQSPAIMSASPGEKVTISCSASSSVSYMYWYQQKPGSSPKPWIYRTSNLASGVPAR

[0144] FSGSGSGTSYSLTISSMEAEDAATYYCQQYHSYPLTFGAGTKLELK

[0145] K64-27 heavy chain variable region (VH) sequence (starting from FR-H1):

[0146] EVKLEESGGGLVQPGGSMKLSCAASGFTFSDAYMDWVRQSPEKGLEWVAEIRSKANN

[0147] HATYYAESVKGRFTISRDDSKSSVYLQMNSLRAEDTGIYYCTPYYYGSSQFAYWGQGT

[0148] LVTVSA

[0149] K64-27 light chain complementarity determining region 1 (LCDR1) sequence:

[0150] SASSSVSYMY

[0151] K64-27 light chain complementarity determining region 2 (LCDR2) sequence:

[0152] RTSNLAS

[0153] K64-27 light chain complementarity determining region 3 (LCDR3) sequence:

[0154] QQYHSYPLT

[0155] Complementarity determining region 1 (HCDR1) sequence of K64-27 heavy chain:

[0156] DAYMD

[0157] Complementarity determining region 2 (HCDR2) sequence of K64-27 heavy chain:

[0158] EIRSKANNHATYYAESVKG

[0159] Complementarity determining region 3 (HCDR3) sequence of K64-27 heavy chain:

[0160] YYYGSSQFAY

[0161] As described in this application, the term "CDR" or "complementarity determining region" refers to the non-contiguous antigen binding sites found in the variable regions of heavy and light chain polypeptides. In this application, the amino acid sequences of the CDRs listed above are all shown according to the Kabat definition rules (the claims of the present invention are also the sequences shown according to the Kabat definition rules). However, it is well known to those skilled in the art that the CDRs of antibodies can be defined in this field by a variety of methods. In the literature Kabat et al., J. Biol. Chem. 252: 6609-6616 (1977); Kabat et al., USDept. of Health and Human Services, "Sequences of proteins of immunological interest" (1991); Chothia et al., J. Mol. Biol. 196: 901-917 (1987); Al-Lazikani B. et al. al., J.Mal.Biol., 273:927-948 (1997); MacCallum et al, J.Mal.Biol.262:732-745 (1996); Abhinandan and Martin, Mal.Immunol., 45:3832-3839 (2008); Lefranc MPet al., Dev. Comp. J. Munol., 27: 55-77 (2003); and Honegger and Plitikthun, J. Mal Biol., 309: 657-670 (2001) have described these special regions, wherein when compared to each other, these definitions include overlaps or subsets of amino acid residues. However, any of the definitions are used to indicate the CDRs of antibodies or transplanted antibodies or their variants, and are included in the scope of the terms defined and used in this application. The algorithms and binding interfaces for CDR prediction are known in the art, such as Abhinandan and Martin, Mal. Immunol., 45: 3832-3839 (2008); Ehrenmann F. et al, Nucleic Acids Res., 38: D301-D307 (2010); and Adolf-Bryfogle J. et al., Nucleic Acids Res., 43: D432-D438 (2015). The contents of the references cited in this paragraph are incorporated into the present application by reference in their entirety for use in the present application and in one or more claims that may be included in the present application.

[0162] As used herein, "monoclonal antibody" refers to a homologous antibody group that participates in the highly specific recognition and binding of a single antigenic determinant or epitope, covering complete and full-length monoclonal antibodies as well as antibody fragments (such as Fab, Fab', F(ab')2, Fv), single-chain (scFv) mutants, fusion proteins comprising antibody portions, and any other modified immunoglobulin molecules comprising antigen recognition sites. In addition, "monoclonal antibody" refers to such antibodies prepared in any number of ways (including but not limited to by hybridoma, phage selection, recombinant expression, and transgenic animals). In humans, this category includes IgG1, IgG2, IgG3, and IgG4. In mice, this category includes IgG1, IgG2a, IgG2b, and IgG3. It is known in the art that the antigen binding function of an antibody can be performed by a fragment of a full-length antibody. The monoclonal antibody or antigen-binding fragment described in the claims of the present invention comprises an Fc fragment in the above-mentioned category of antibodies.

[0163] 2) Preparation and purification of monoclonal antibodies against Klebsiella pneumoniae K64 capsular polysaccharide in ascites

[0164] The monoclonal antibody cells obtained were expanded and cultured, and 0.5 mL of 1×10 6 100 cells / mL were added to BALB / c mice sensitized with paraffin oil 7 days in advance. After 7-10 days, the abdomen of the mice was observed to have obvious bulges, and the ascites was collected for purification.

[0165] The mouse ascites was diluted with PBS, and 50% final concentration of ammonium sulfate was added for salt precipitation and then purified by Protein A affinity column chromatography and ion exchange chromatography to obtain a monoclonal antibody with a purity greater than 95%. After sterile filtration through a 0.22 μm filter membrane, it was stored in PBS at pH 7.0-7.4.

[0166] Example 2: Titer level detection of monoclonal antibodies to Klebsiella pneumoniae K64 capsular polysaccharide

[0167] After cell fusion, screening and several subclones, 7 monoclonal antibody cell lines were obtained. The results of ELISA test on the binding of cell culture supernatants to K64 type capsular polysaccharide are shown in Table 1 below.

[0168] Table 1

[0169] Cell line number OD value K64-1 3.816 K64-23 1.315 K64-24 2.011 K64-3 3.227 K64-5 0.707 K64-8 3.362 K64-27 1.043

[0170] like Figure 1 As shown, the titer level of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody provided in the present application was detected by indirect ELISA method, and it can be seen that the supernatants of the seven cell strains all have good binding affinity with the K64 type capsular polysaccharide.

[0171] Example 3: ELISA detection of binding specificity of monoclonal antibodies to Klebsiella pneumoniae K64 capsular polysaccharide

[0172] The monoclonal antibody of Klebsiella pneumoniae capsular polysaccharide provided in this application was analyzed by indirect ELISA method to react with K47, K64, K19, K1, K2, K38 and K57 capsular polysaccharides to evaluate its specificity. The Klebsiella pneumoniae capsular polysaccharide of different serotypes was coated in an ELISA plate at 37°C for 3 hours. After blocking with 1% BSA for 1 hour, an appropriate amount of K64 Klebsiella pneumoniae capsular polysaccharide monoclonal antibody was added to react with the coated polysaccharide, 50 μL per well, and incubated overnight. After washing the plate, the AP-labeled goat anti-mouse secondary antibody was diluted 1:30,000 and added to the ELISA plate, 100 μL per well, incubated for 2 hours, and then washed, and then 1 mg / mL of PNPP-Na colorimetric substrate was added, 100 μL per well, and after incubation for 2 hours, 3M NaOH was added to 50 μL wells to stop the reaction, and the OD405 value was read on the machine.

[0173] The results of the binding of the 7 K64 type Klebsiella pneumoniae capsular polysaccharide monoclonal antibodies provided in the present application to the Klebsiella capsular polysaccharides of different serotypes are shown in Table 2 below:

[0174] Table 2

[0175]

[0176]

[0177] It can be seen from Table 2 above that the 7 monoclonal antibodies against K64-type Klebsiella pneumoniae capsular polysaccharide provided in the present application are specific to K64-type capsular polysaccharide and have no cross-reaction with other serotypes of Klebsiella capsular polysaccharide.

[0178] Example 4: Flow cytometry detection of binding between monoclonal antibody against Klebsiella pneumoniae K64 capsular polysaccharide and Klebsiella pneumoniae K64

[0179] K64-Serotype highly virulent Klebsiella pneumoniae Y8 (Li, J.; Sheng, Y.; Ma, R.; Xu, M.; Liu, F.; Qin, R.; Zhu, M.; Zhu, X.; He, P. Identification of a Depolymerase Specific for K64-Serotype Klebsiella pneumoniae: Potential Applications in Capsular Typing and Treatment. Antibiotics 2021, 10, 144.) was inoculated into 5 ml LB medium and cultured at 37°C and 250 rpm until the OD value was 0.7. 0.2 mL of the bacterial solution was taken and centrifuged at 8000 g for 10 min. The precipitate was collected, washed twice with PBS, and resuspended in 0.2 mL PBS; then fixed with 4% paraformaldehyde for 10 minutes, blocked with 2% BSA for 1 hour after washing, and then incubated with K64 capsular polysaccharide monoclonal antibody overnight, added with anti-mouse fluorescent antibody (brand-CST, item number-#4410S) after washing, incubated in the dark for 1 hour, washed with PBS, resuspended in PBS and detected by flow cytometry.

[0180] After culturing, the highly virulent strain Y8 of Klebsiella pneumoniae was combined with the K64 monoclonal antibody and the binding ability was analyzed by flow cytometry. The test results are as follows Figure 2 As shown in the figure, seven K64-type monoclonal antibodies were tested for binding to the K64-type highly virulent Klebsiella pneumoniae strain Y8. The results showed that all seven K64-type monoclonal antibodies could bind to Y8, while the negative control did not bind to Y8, indicating that these seven K64-type capsular polysaccharide monoclonal antibodies can be used for the diagnosis and typing of clinical strains.

[0181] Example 5: Bactericidal test of monoclonal antibody against Klebsiella pneumoniae K64 capsular polysaccharide

[0182] This example shows the opsonization and bactericidal test of the monoclonal antibody of Klebsiella pneumoniae capsular polysaccharide provided by the present application, specifically: the K64 type highly virulent Klebsiella pneumoniae Y8 strain was diluted 10 5 CFU / ml, and added to the 96-well cell working plate at 10 μL / well. K64-type capsular polysaccharide monoclonal antibody was diluted in gradients and added to the above cell working plate at 20 μL / well. The bacteria and antibodies were incubated at 700 rpm / min for 30 min. The HL-60 cells differentiated by DMF were washed with HBSS buffer and adjusted to a concentration of 1×10 71×107 cells / ml, then mix the cell solution of 1×107 cells / ml with the diluted complement in a volume ratio of 1:4, and add 50μL of the mixture to the 96-well cell plate. Place the 96-well cell plate on a mixer, put it in a CO2 incubator with 5% CO2 and a temperature of 37°C, and shake and culture for 45 minutes. After terminating the opsonized phagocytosis, spot the sample on the blood plate and culture it in a CO2 incubator overnight. Calculate the bactericidal rate of each diluted antibody.

[0183] like Figure 3 As shown, K64-type capsular polysaccharide monoclonal antibodies K64-1, K64-23, and K64-24 had good bactericidal effects on the highly virulent strain Y8 in vitro, with the highest bactericidal rates reaching 83.3% and 86.7%, respectively, which were significantly different from the negative control antibodies.

[0184] Example 6: Preventive test of monoclonal antibodies against Klebsiella pneumoniae K64 capsular polysaccharide

[0185] This example shows the protective test of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody provided by the present application, specifically: K64-1, a Klebsiella pneumoniae capsular polysaccharide monoclonal antibody, was administered by intraperitoneal injection to 6 mice in each group, with each mouse receiving 0.025 mg, and PBS was used as a negative control. After 4 hours, the highly virulent strain Y8 of Klebsiella pneumoniae was injected at 7.38×10 6 The amount of CFU was used to infect Balb / c mice (n=6) intraperitoneally. The survival rate of the mice was observed for 10 days after infection, and a survival curve was drawn.

[0186] like Figure 4 As shown in the figure, it can be seen that the monoclonal antibody K64-1 against K64 capsular polysaccharide and the control PBS administration 4 hours later reduced the amount of Klebsiella pneumoniae type 64 (strain number Y8) by 7.38×10 6 After CFU challenge, the survival rate of the control group was 0%, while that of the monoclonal antibody group was 100%, with statistically significant difference (p<0.05).

[0187] In summary, the above embodiments are only preferred embodiments of the present application and are not intended to limit the protection scope of the present application. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. A monoclonal antibody against Klebsiella pneumoniae capsular polysaccharide, characterized in that: The Klebsiella pneumoniae is K64 type Klebsiella pneumoniae, and the sequence of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody comprises a light chain variable region and a heavy chain variable region, the amino acid sequence of the light chain variable region is as shown in SEQ NO:1, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto, and the amino acid sequence of the heavy chain variable region is as shown in SEQ NO:2, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto.

2. The Klebsiella pneumoniae capsular polysaccharide monoclonal antibody according to claim 1, characterized in that: The complementary determining region of the light chain variable region comprises three sequences as shown in SEQ ID:3, SEQ ID:4 and SEQ ID:5, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:3, SEQ ID:4 and SEQ ID:5; the complementary determining region of the heavy chain variable region comprises three sequences as shown in SEQ ID:6, SEQ ID:7 and SEQ ID:8, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:6, SEQ ID:7 and SEQ ID:

8.

3. A hybridoma cell line, characterized in that: The hybridoma cell line is named K64-1, the deposit number of the hybridoma cell line is CCTCC NO: C2023365, and the hybridoma cell line is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody as claimed in claim 1.

4. A monoclonal antibody against Klebsiella pneumoniae capsular polysaccharide, characterized in that: The Klebsiella pneumoniae is K64 type Klebsiella pneumoniae, and the sequence of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody comprises a light chain variable region and a heavy chain variable region, the amino acid sequence of the light chain variable region is as shown in SEQ NO:9 or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto, and the amino acid sequence of the heavy chain variable region is as shown in SEQ NO:10 or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto.

5. The Klebsiella pneumoniae capsular polysaccharide monoclonal antibody according to claim 4, characterized in that: The complementarity determining region of the light chain variable region comprises three sequences as shown in SEQ ID:11, SEQ ID:12 and SEQ ID:13, or sequences having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:11, SEQ ID:12 and SEQ ID:13; the complementarity determining region of the heavy chain variable region comprises three sequences as shown in SEQ ID:14, SEQ ID:15 and SEQ ID:16, or sequences having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:14, SEQ ID:15 and SEQ ID:

16.

6. A hybridoma cell line, characterized in that: The hybridoma cell line is named K64-23, the deposit number of the hybridoma cell line is CCTCC NO: C2023366, and the hybridoma cell line is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody as claimed in claim 4.

7. A monoclonal antibody against Klebsiella pneumoniae capsular polysaccharide, characterized in that: The Klebsiella pneumoniae is K64 type Klebsiella pneumoniae, and the sequence of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody comprises a light chain variable region and a heavy chain variable region, the amino acid sequence of the light chain variable region is as shown in SEQ NO:17, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto, and the amino acid sequence of the heavy chain variable region is as shown in SEQ NO:18, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto.

8. The Klebsiella pneumoniae capsular polysaccharide monoclonal antibody according to claim 7, characterized in that: The complementarity determining region of the light chain variable region comprises three sequences as shown in SEQ ID: 19, SEQ ID: 20 and SEQ ID: 21, or sequences having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID: 19, SEQ ID: 20 and SEQ ID: 21, and the complementarity determining region of the heavy chain variable region comprises three sequences as shown in SEQ ID: 22, SEQ ID: 23 and SEQ ID: 24, or sequences having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID: 22, SEQ ID: 23 and SEQ ID:

24.

9. A hybridoma cell line, characterized in that: The hybridoma cell line is named K64-24, the deposit number of the hybridoma cell line is CCTCC NO: C2023367, and the hybridoma cell line is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody as claimed in claim 7.

10. A monoclonal antibody against Klebsiella pneumoniae capsular polysaccharide, characterized in that: The Klebsiella pneumoniae is K64 type Klebsiella pneumoniae, and the sequence of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody comprises a light chain variable region and a heavy chain variable region, the amino acid sequence of the light chain variable region is as shown in SEQ NO:25, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto, and the amino acid sequence of the heavy chain variable region is as shown in SEQ NO:26, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto.

11. The Klebsiella pneumoniae capsular polysaccharide monoclonal antibody according to claim 10, characterized in that: The complementarity determining region of the light chain variable region comprises three sequences as shown in SEQ ID:27, SEQ ID:28 and SEQ ID:29, or sequences having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:27, SEQ ID:28 and SEQ ID:29, and the complementarity determining region of the heavy chain variable region comprises three sequences as shown in SEQ ID:30, SEQ ID:31 and SEQ ID:32, or sequences having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:30, SEQ ID:31 and SEQ ID:

32.

12. A hybridoma cell line, characterized in that: The hybridoma cell line is named K64-3, the deposit number of the hybridoma cell line is CCTCC NO: C2024340, and the hybridoma cell line is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody as claimed in claim 10.

13. A monoclonal antibody against Klebsiella pneumoniae capsular polysaccharide, characterized in that: The Klebsiella pneumoniae is K64 type Klebsiella pneumoniae, and the sequence of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody comprises a light chain variable region and a heavy chain variable region, the amino acid sequence of the light chain variable region is as shown in SEQ NO:33, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto, and the amino acid sequence of the heavy chain variable region is as shown in SEQ NO:34, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto.

14. The Klebsiella pneumoniae capsular polysaccharide monoclonal antibody according to claim 13, characterized in that: The complementarity determining region of the light chain variable region comprises three sequences as shown in SEQ ID:35, SEQ ID:36 and SEQ ID:37, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:35, SEQ ID:36 and SEQ ID:37; the complementarity determining region of the heavy chain variable region comprises three sequences as shown in SEQ ID:38, SEQ ID:39 and SEQ ID:40, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:38, SEQ ID:39 and SEQ ID:

40.

15. A hybridoma cell line, characterized in that: The hybridoma cell line is named K64-5, the deposit number of the hybridoma cell line is CCTCC NO: C2024341, and the hybridoma cell line is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody as claimed in claim 13.

16. A monoclonal antibody against Klebsiella pneumoniae capsular polysaccharide, characterized in that: The Klebsiella pneumoniae is K64 type Klebsiella pneumoniae, and the sequence of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody comprises a light chain variable region and a heavy chain variable region, the amino acid sequence of the light chain variable region is as shown in SEQ NO:41, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto, and the amino acid sequence of the heavy chain variable region is as shown in SEQ NO:42, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto.

17. The Klebsiella pneumoniae capsular polysaccharide monoclonal antibody according to claim 16, characterized in that: The complementarity determining region of the light chain variable region comprises three sequences as shown in SEQ ID:43, SEQ ID:44 and SEQ ID:45, or sequences having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:43, SEQ ID:44 and SEQ ID:45, and the complementarity determining region of the heavy chain variable region comprises three sequences as shown in SEQ ID:46, SEQ ID:47 and SEQ ID:48, or sequences having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:46, SEQ ID:47 and SEQ ID:

48.

18. A hybridoma cell line, characterized in that: The hybridoma cell line is named K64-8, the deposit number of the hybridoma cell line is CCTCC NO: C2024342, and the hybridoma cell line is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody according to claim 16.

19. A monoclonal antibody against Klebsiella pneumoniae capsular polysaccharide, characterized in that: The Klebsiella pneumoniae is K64 type Klebsiella pneumoniae, and the sequence of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody comprises a light chain variable region and a heavy chain variable region, the amino acid sequence of the light chain variable region is as shown in SEQ NO:49, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto, and the amino acid sequence of the heavy chain variable region is as shown in SEQ NO:50, or a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto.

20. The Klebsiella pneumoniae capsular polysaccharide monoclonal antibody according to claim 19, characterized in that: The complementarity determining region of the light chain variable region comprises three sequences as shown in SEQ ID:51, SEQ ID:52 and SEQ ID:53, or sequences having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:51, SEQ ID:52 and SEQ ID:53; the complementarity determining region of the heavy chain variable region comprises three sequences as shown in SEQ ID:54, SEQ ID:55 and SEQ ID:56, or sequences having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity compared to any one of SEQ ID:54, SEQ ID:55 and SEQ ID:

56.

21. A hybridoma cell line, characterized in that: The hybridoma cell line is named K64-27, the deposit number of the hybridoma cell line is CCTCC NO: C2024343, and the hybridoma cell line is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody as claimed in claim 19.

22. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody according to any one of claims 1, 4, 7, 10, 13, 16 and 19, and further comprises any one or more combinations of pharmaceutically acceptable carriers, excipients or diluents.

23. A kit for detecting K64 type Klebsiella pneumoniae, characterized in that: The kit for detecting K64 type Klebsiella pneumoniae comprises the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody described in any one of claims 1, 4, 7, 10, 13, 16 and 19, and the kit for detecting K64 type Klebsiella pneumoniae further comprises any one or more combinations of a positive control, a negative control, an antibody diluent, a color developing solution, a stop solution, a blocking solution or a washing solution.

24. Use of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody according to any one of claims 1, 4, 7, 10, 13, 16 and 19 in the preparation of a drug for treating a K64 type Klebsiella pneumoniae infection disease and / or a detection product.

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