K47-type klebsiella pneumoniae capsular polysaccharide monoclonal antibody, hybridoma cell lines therefor and use thereof

Abstract

The present application discloses a K47-type klebsiella pneumoniae capsular polysaccharide monoclonal antibody, hybridoma cell lines therefor and a use thereof. The sequence information of the K47-type klebsiella pneumoniae capsular polysaccharide monoclonal antibody is shown in SEQ NO: 1 to SEQ NO: 48; and the preservation numbers of the hybridoma cell lines are CCTCC:C2023296, CCTCC:C2023311, CCTCC:C2023312, CCTCC:C2023313, CCTCC:C2023314 and CCTCC:C2024339, respectively. The K47-type klebsiella pneumoniae capsular polysaccharide monoclonal antibody provided by the present application can specifically bind to capsular polysaccharide of klebsiella pneumoniae, is beneficial to typing of klebsiella pneumoniae in clinic settings, and can be proved in both animal in-vivo and in-vitro experiments to have excellent efficacy in preventing and treating K47-type klebsiella pneumoniae infections; thus, the K47-type klebsiella pneumoniae capsular polysaccharide monoclonal antibody provides a way to solve the problems of klebsiella pneumoniae infections and multi-drug resistance, is of remarkable significance for the prevention, diagnosis and treatment of klebsiella pneumoniae infections, and has important value for the development of new-generation drugs against klebsiella pneumoniae.

Description

K47 Klebsiella pneumoniae capsular polysaccharide monoclonal antibody, hybridoma cell line thereof and application thereof Technical Field

[0001] The present application belongs to the field of medical technology, and in particular relates to a monoclonal antibody against Klebsiella pneumoniae capsular polysaccharide of type K47, a hybridoma cell line thereof, and applications thereof. Background Art

[0002] Klebsiella pneumoniae is a Gram-negative (G-) bacterium with an encapsulated bacterium. It primarily colonizes the digestive and respiratory tracts. When the body's immune system is weakened, it can cause lung infections, urinary tract infections, and even bloodstream infections. It is also one of the most common bacterial causes of respiratory tract infections.

[0003] Infections caused by Klebsiella pneumoniae have become a serious global public health problem. Statistics show that Klebsiella pneumoniae infections have an extremely high mortality rate, ranging from 20% to 30% for bloodstream infections and over 50% for bacteremia combined with pneumonia. Treatment of Klebsiella pneumoniae primarily relies on antibiotics. However, with the widespread use of various antimicrobial agents in recent years, Klebsiella pneumoniae that produce extended-spectrum β-lactamases and carbapenemases have become resistant to nearly all available β-lactams, including carbapenems. Over the past decade, the incidence of carbapenem-resistant Klebsiella pneumoniae (CRKP) has increased dramatically worldwide. Capsular polysaccharides are the primary virulence factor of Klebsiella pneumoniae and serve as important protective antigens for the treatment or prevention of Klebsiella pneumoniae infections. Based on the structure of the capsular polysaccharide, Klebsiella pneumoniae can be divided into at least 77 serotypes, also known as K antigens or K types. Different K types exhibit distinct resistance profiles and differ in geographic and clinical distribution, potentially leading to different epidemic patterns in hospital-acquired infections, thus directly impacting clinical treatment options. Among them, K47 type Klebsiella pneumoniae is often associated with more severe infections in clinical practice, particularly in immunosuppressed patients.

[0004] Therefore, there is a need in this field to develop a K47 type Klebsiella pneumoniae capsular polysaccharide monoclonal antibody with high specificity, good affinity and strong binding ability, so as to facilitate 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 K47 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 complementarity determining regions of the light chain variable region comprise three sequences as shown in SEQ ID: 3, SEQ ID: 4 and SEQ ID: 5, 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: 3, SEQ ID: 4 and SEQ ID: 5; the complementarity determining regions of the heavy chain variable region comprise three sequences as shown in SEQ ID: 6, SEQ ID: 7 and SEQ ID: 8, 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: 6, SEQ ID: 7 and SEQ ID: 8.

[0007] In a second aspect, the present application provides a hybridoma cell line, which is named K47-5 B9-F8 and has a deposit number of CCTCC NO: C2023314. The hybridoma cell line is used to prepare the aforementioned Klebsiella pneumoniae capsular polysaccharide monoclonal antibody.

[0008] In a third aspect, the present application provides a Klebsiella pneumoniae capsular polysaccharide monoclonal antibody, wherein the Klebsiella pneumoniae is K47 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 regions of the light chain variable region comprise 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 regions of the heavy chain variable region comprise 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.

[0010] In a fourth aspect, the present application provides a hybridoma cell line, which is named K47-15D7-B10, and the deposit number of the hybridoma cell line is: CCTCC NO: C2023312. The hybridoma cell line is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody 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 K47 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 regions of the light chain variable region comprise 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 regions of the heavy chain variable region comprise 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.

[0013] In a sixth aspect, the present application provides a hybridoma cell line, which is named K47-18G6-B2, and the deposit number of the hybridoma cell line is: CCTCC NO: C2023311. The hybridoma cell line 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 K47 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 regions of the light chain variable region comprise 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 regions of the heavy chain variable region comprise 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.

[0016] In an eighth aspect, the present application provides a hybridoma cell line, which is named K47-6E2 and has a deposit number of CCTCC NO: C2023296. The hybridoma cell line is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody as described in the seventh aspect.

[0017] Ninth aspect, the present application provides a Klebsiella pneumoniae capsular polysaccharide monoclonal antibody, wherein the Klebsiella pneumoniae is K47 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 regions of the light chain variable region comprise three sequences as shown in SEQ ID: 35, SEQ ID: 36 and SEQ ID: 37, 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: 35, SEQ ID: 36 and SEQ ID: 37, and the complementarity determining regions of the heavy chain variable region comprise three sequences as shown in SEQ ID: 38, SEQ ID: 39 and SEQ ID: 40, 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: 38, SEQ ID: 39 and SEQ ID: 40.

[0019] In the tenth aspect, the present application provides a hybridoma cell line, the hybridoma cell line is named K47-20C10-B8, the deposit number of the hybridoma cell line is: CCTCC NO: C2023313, and the hybridoma cell line is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody as described in the ninth aspect.

[0020] In an eleventh aspect, the present application provides a Klebsiella pneumoniae capsular polysaccharide monoclonal antibody, wherein the Klebsiella pneumoniae is K47 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 regions of the light chain variable region comprise 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 regions of the heavy chain variable region comprise 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.

[0022] In the twelfth aspect, the present application provides a hybridoma cell line, which is named K47-4H8, and the deposit number of the hybridoma cell line is: CCTCC: C2024339. The hybridoma cell line 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 pharmaceutical composition, comprising the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody described in any one 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.

[0024] In a fourteenth aspect, the present application provides a kit for detecting K47 type Klebsiella pneumoniae, wherein the kit for detecting K47 type Klebsiella pneumoniae comprises the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody described in any one of the aforementioned methods, and the kit for detecting K47 type Klebsiella pneumoniae further comprises a positive control, a negative control, an antibody diluent, a color developing solution, a stopping solution, a blocking solution or a washing solution, in combination with any one or more of the following.

[0025] In a fifteenth aspect, the present application provides the use of any of the aforementioned Klebsiella pneumoniae capsular polysaccharide monoclonal antibodies in the preparation of therapeutic drugs and / or detection products for Klebsiella pneumoniae infection diseases, 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.

[0026] Capsular polysaccharide is an important toxic factor of Klebsiella pneumoniae. This application uses capsular polysaccharide to couple with carrier protein to improve immunogenicity and prepares K47 type Klebsiella pneumoniae capsular polysaccharide-specific monoclonal antibodies using hybridoma technology. The antibodies have good specificity, high affinity, and high binding ability to Klebsiella pneumoniae capsular polysaccharide.

[0027] The beneficial effects of this application are:

[0028] The Klebsiella pneumoniae capsular polysaccharide monoclonal antibody provided in the present application can specifically bind to the capsular polysaccharide of Klebsiella pneumoniae type K47, which is beneficial for the typing of Klebsiella pneumoniae in clinical practice. It can be demonstrated in both in vivo and in vitro animal experiments that it has excellent effects in preventing and treating Klebsiella pneumoniae infection. It 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

[0029] FIG1 is a graph showing the titer of the monoclonal antibody against Klebsiella pneumoniae capsular polysaccharide of K47 provided by the present application detected by indirect ELISA in Example 2;

[0030] Figure 2 is a flow cytometric analysis of the binding of monoclonal antibody K47-20C10-B8 to the encapsulated Klebsiella pneumoniae HVKP4 and the non-encapsulated Klebsiella pneumoniae 19-249ΔwbaP (capsular polysaccharide expression gene knockout) in Example 4;

[0031] FIG3 is a flow cytometric analysis of the binding of monoclonal antibody K47-18G6-B2 to the encapsulated Klebsiella pneumoniae HVKP4 and the non-encapsulated Klebsiella pneumoniae 19-249ΔwbaP (capsular polysaccharide expression gene knockout) in Example 4;

[0032] FIG4 is a bactericidal curve of K47-5B9-F8 and K47-15D7-B10 against Klebsiella pneumoniae HVKP4 strain in vitro in Example 5;

[0033] FIG5 is a survival curve of the prevention test of the monoclonal antibody against Klebsiella pneumoniae capsular polysaccharide K47 provided by the present application in Example 6. DETAILED DESCRIPTION

[0034] The technical scheme of the present application will be clearly and completely described below. Obviously, the described embodiments are some embodiments of the present application, rather than all embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without making creative work fall within the scope of protection of the present application. The following examples relate to reagents that are commercially available in this area unless otherwise specified.

[0035] The following is the strain deposit information:

[0036] The hybridoma cell line K47-5B9-F8 (CCTCC NO: C2023314), hybridoma cell line K47-20C10-B8 (CCTCC NO: C2023313), hybridoma cell line K47-15D7-B10 (CCTCC NO: C2023312), hybridoma cell line K47-18G6-B2 (CCTCC NO: C2023311) and hybridoma cell line K47-6E2 (CCTCC NO: C2023296) provided in this application have all been deposited in the China Center for Type Culture Collection on October 17, 2023.

[0037] The hybridoma cell line K47-4H8 (CCTCC: C2024339) provided in this application has been deposited in the China Center for Type Culture Collection on September 25, 2024.

[0038] Deposit address: Wuhan University, Wuhan, Hubei Province.

[0039] Deposit name and deposit number:

[0040] Hybridoma cell line K47-5B9-F8: Hybridoma cell line K47-5B9-F8, CCTCC NO: C2023314.

[0041] Hybridoma cell line K47-20C10-B8: Hybridoma cell line K47-20C10-B8, CCTCC NO: C2023313.

[0042] Hybridoma cell line K47-15D7-B10: Hybridoma cell line K47-15D7-B10, CCTCC NO: C2023312.

[0043] Hybridoma cell line K47-18G6-B2: Hybridoma cell line K47-18G6-B2, CCTCC NO: C2023311.

[0044] Hybridoma cell line K47-6E2: Hybridoma cell line K47-6E2, CCTCC NO: C2023296.

[0045] Hybridoma cell line K47-4H8: Hybridoma cell line K47-6E2, CCTCC: C2024339.

[0046] In Example 5 of the present application, the light chain variable region (VL) sequence of the negative control antibody used (as shown in SEQ ID NO: 49) is:

[0047]

[0048] The negative control antibody heavy chain variable region (VH) sequence used (as shown in SEQ ID NO: 50):

[0049]

[0050] Example 1: Preparation of monoclonal antibodies against Klebsiella pneumoniae K47 capsular polysaccharide

[0051] 1) Establishment of a cell line producing monoclonal antibodies against Klebsiella pneumoniae K47 capsular polysaccharide

[0052] The K47 capsular polysaccharide-protein conjugate was obtained by chemically coupling the capsular polysaccharide extracted from the fermentation of Klebsiella pneumoniae K47 (GDMCC No: 62131) with a carrier protein (diphtheria toxin-free variant CRM197) using commonly used polysaccharide activation methods, such as the cyanogen bromide method (reference US6375846B1), 1-cyano-4-dimethylamino arsenic tetrafluoroborate (CDAP) (EP0720485), and the periodate oxidation method (US4711779). The K47 capsular polysaccharide protein conjugate was added to Freund's adjuvant and immunized into SPF-grade BALB / c mice multiple times. When the immune titer was greater than 1:300,000, the spleens were harvested and hybridoma technology was used. After PEG1500 fusion, indirect ELISA screening of positive cell supernatants, and multiple subcloning, five monoclonal cell lines were obtained, namely, hybridoma cell line K47-5B9-F8 (CCTCC NO: C2023314), hybridoma cell line K47-20C10-B8 (CCTCC NO: C2023313), hybridoma cell line K47-15D7-B10 (CCTCC NO: C2023312), hybridoma cell line K47-18G6-B2 (CCTCC NO: C2023311), hybridoma cell line K47-6E2 (CCTCC NO: C2023313), and hybridoma cell line K47-15D7-B10 (CCTCC NO: C2023312). NO: C2023296) and hybridoma cell line K47-4H8 (CCTCC: C2024339), and their purity was verified by sequencing.

[0053] After sequencing, the sequence information of 6 Klebsiella pneumoniae capsular polysaccharide monoclonal antibodies are as follows (SEQ NO: 1 to SEQ NO: 48):

[0054] K47-5B9-F8-light chain variable region (VL) sequence:

[0055]

[0056] K47-5B9-F8-heavy chain variable region (VH) sequence:

[0057]

[0058] K47-5B9-F8 - light chain complementarity determining region 1 (LCDR1) sequence:

[0059]

[0060] K47-5B9-F8 - light chain complementarity determining region 2 (LCDR2) sequence:

[0061]

[0062] K47-5B9-F8 - light chain complementarity determining region 3 (LCDR3) sequence:

[0063]

[0064] K47-5B9-F8 - heavy chain complementarity determining region 1 (HCDR1) sequence:

[0065]

[0066] K47-5B9-F8 - heavy chain complementarity determining region 2 (HCDR2) sequence:

[0067]

[0068] K47-5B9-F8 - heavy chain complementarity determining region 3 (HCDR3) sequence:

[0069]

[0070] K47-15D7-B10-light chain variable region (VL) sequence:

[0071]

[0072] K47-15D7-B10-heavy chain variable region (VH) sequence:

[0073]

[0074] K47-15D7-B10 - light chain complementarity determining region 1 (LCDR1) sequence:

[0075]

[0076] K47-15D7-B10 - light chain complementarity determining region 2 (LCDR2) sequence:

[0077]

[0078] K47-15D7-B10 - light chain complementarity determining region 3 (LCDR3) sequence:

[0079]

[0080] K47-15D7-B10 - heavy chain complementarity determining region 1 (HCDR1) sequence:

[0081]

[0082] K47-15D7-B10 - heavy chain complementarity determining region 2 (HCDR2) sequence:

[0083]

[0084] K47-15D7-B10 - heavy chain complementarity determining region 3 (HCDR3) sequence:

[0085]

[0086] K47-18G6-B2-light chain variable region (VL) sequence:

[0087]

[0088] K47-18G6-B2-heavy chain variable region (VH) sequence:

[0089]

[0090] K47-18G6-B2 - light chain complementarity determining region 1 (LCDR1) sequence:

[0091]

[0092] K47-18G6-B2 - light chain complementarity determining region 2 (LCDR2) sequence:

[0093]

[0094] K47-18G6-B2 - light chain complementarity determining region 3 (LCDR3) sequence:

[0095]

[0096] K47-18G6-B2 - heavy chain complementarity determining region 1 (HCDR1) sequence:

[0097]

[0098] K47-18G6-B2 - heavy chain complementarity determining region 2 (HCDR2) sequence:

[0099]

[0100] K47-18G6-B2- heavy chain complementarity determining region 3 (HCDR3) sequence:

[0101]

[0102] K47-6E2-light chain variable region (VL) sequence:

[0103]

[0104] K47-6E2-heavy chain variable region (VH) sequence:

[0105]

[0106] K47-6E2-light chain complementarity determining region 1 (LCDR1) sequence:

[0107]

[0108] K47-6E2-light chain complementarity determining region 2 (LCDR2) sequence:

[0109]

[0110] K47-6E2-light chain complementarity determining region 3 (LCDR3) sequence:

[0111]

[0112] K47-6E2- heavy chain complementarity determining region 1 (HCDR1) sequence:

[0113]

[0114] K47-6E2- heavy chain complementarity determining region 2 (HCDR2) sequence:

[0115]

[0116] K47-6E2- heavy chain complementarity determining region 3 (HCDR3) sequence:

[0117]

[0118] K47-20C10-B8-light chain variable region (VL) sequence:

[0119]

[0120] K47-20C10-B8-heavy chain variable region (VH) sequence:

[0121]

[0122] K47-20C10-B8 - light chain complementarity determining region 1 (LCDR1) sequence:

[0123]

[0124] K47-20C10-B8 - light chain complementarity determining region 2 (LCDR2) sequence:

[0125]

[0126] K47-20C10-B8 - light chain complementarity determining region 3 (LCDR3) sequence:

[0127]

[0128] K47-20C10-B8 - heavy chain complementarity determining region 1 (HCDR1) sequence:

[0129]

[0130] K47-20C10-B8 - heavy chain complementarity determining region 2 (HCDR2) sequence:

[0131]

[0132] K47-20C10-B8 - heavy chain complementarity determining region 3 (HCDR3) sequence:

[0133]

[0134] K47-4H8-light chain variable region (VL) sequence:

[0135]

[0136] K47-4H8-heavy chain variable region (VH) sequence:

[0137]

[0138] K47-4H8-light chain complementarity determining region 1 (LCDR1) sequence:

[0139]

[0140] K47-4H8-light chain complementarity determining region 2 (LCDR2) sequence:

[0141]

[0142] K47-4H8-light chain complementarity determining region 3 (LCDR3) sequence:

[0143]

[0144] K47-4H8- heavy chain complementarity determining region 1 (HCDR1) sequence:

[0145]

[0146] K47-4H8-Complementarity Determining Region 2 (HCDR2) sequence of the heavy chain:

[0147]

[0148] K47-4H8- heavy chain complementarity determining region 3 (HCDR3) sequence:

[0149]

[0150] As used herein, the term "CDR" or "complementarity determining region" refers to the non-contiguous antigen binding sites found within 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 (the claims of the present invention also show sequences according to the Kabat definition). However, it is well known in the art that antibody CDRs can be defined in the art using 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 These specific regions have been described in, for example, Desmond et al., Dev. Comp. J. Am. Biol., 27:55-77 (2003); and Honegger and Plitikthun, J. Mal. Biol., 309:657-670 (2001), wherein these definitions include overlap or subsets of amino acid residues when compared to one another. However, any of the definitions used to designate a CDR of an antibody or grafted antibody or variant thereof is included within the scope of the term as defined and used herein. 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 herein by reference in their entirety for use in this application and in one or more claims that may be included in this application.

[0151] As used herein, "monoclonal antibody" refers to a group of homologous antibodies that participate 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 hybridomas, 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 fragments of a full-length antibody. The monoclonal antibodies or antigen-binding fragments described in the claims of the present invention include Fc fragments in the above-mentioned categories of antibodies.

[0152] 2) Preparation and purification of monoclonal antibodies against Klebsiella pneumoniae K47 polysaccharide in ascites

[0153] The monoclonal antibody cells were expanded and cultured, and 0.5 mL of the monoclonal antibody with a concentration 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 mice were observed to have a noticeable bulge in the abdomen, and the ascites was collected for purification.

[0154] The mouse ascites was diluted with PBS, and 50% final concentration of ammonium sulfate was added to precipitate the crude product. The monoclonal antibody with a purity greater than 90% was obtained after Protein A affinity column chromatography and ion exchange chromatography. The monoclonal antibody was sterile filtered through a 0.22 μm filter membrane and stored in PBS at pH 7.0-7.4.

[0155] Example 2: Titer Level Detection of Monoclonal Antibodies to Klebsiella pneumoniae K47 Capsular Polysaccharide

[0156] After cell fusion, screening, and several subcloning steps, six monoclonal antibody cell lines were obtained. The results of ELISA tests for binding of their cell culture supernatants to K47 capsular polysaccharide are shown in Table 1 below.

[0157] Table 1

[0158]

[0159] As shown in FIG1 , the titer levels of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibodies provided in the present application were detected by indirect ELISA method, and it can be seen that all six monoclonal antibodies have good binding affinity to K47 capsular polysaccharide.

[0160] Example 3: ELISA detection of binding specificity of monoclonal antibodies to Klebsiella pneumoniae K47 capsular polysaccharide

[0161] The Klebsiella pneumoniae capsular polysaccharide monoclonal antibody provided in this application was analyzed by indirect ELISA to react with K47, K64, K19, K1, K2, K38 and K57 capsular polysaccharides to evaluate its specificity. Klebsiella pneumoniae capsular polysaccharides of different serotypes were coated on an enzyme-labeled plate at 37°C for 3 hours. After blocking with 1% BSA for 1 hour, an appropriate amount of K47 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 secondary antibody was diluted 1:30,000 and added to the enzyme-labeled plate, 100 μL per well, incubated for 2 hours, and then washed. Then, 1 mg / mL 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.

[0162] The results of the binding of the six Klebsiella pneumoniae capsular polysaccharide monoclonal antibodies provided in this application to Klebsiella capsular polysaccharides of different serotypes are shown in Table 2 below:

[0163] [Incorporated by reference (Rule 20.6) 24.02.2025] Table 2

[0164] [Incorporated by reference (Rule 20.6) 24.02.2025]

[0165] [Added by reference (Rule 20.6) 24.02.2025] It can be seen from Table 2 above that the six Klebsiella pneumoniae capsular polysaccharide monoclonal antibodies provided in this application are specific for K47 type capsular polysaccharide, and have no cross-reaction with other serotypes of Klebsiella capsular polysaccharides.

[0166] [Incorporated by reference (Rule 20.6) 24.02.2025] Example 4: Flow Cytometry Detection of Binding of Klebsiella pneumoniae K47 Capsular Polysaccharide Monoclonal Antibodies to Klebsiella pneumoniae K47

[0167] [Added by reference (rule 20.6) 24.02.2025] The highly virulent Klebsiella pneumoniae strain HVKP4 and 19-249ΔwbaP strain (capsular polysaccharide expression gene knockout) were inoculated into 5 mL of 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 centrifuged at 8000 g for 10 minutes, the precipitate was collected, washed twice with PBS, and resuspended in 0.2 mL of PBS; then fixed with 4% paraformaldehyde for 10 minutes, washed, and blocked with 2% BSA for 1 hour, and then incubated with K47 type capsular polysaccharide monoclonal antibody overnight. After washing, anti-mouse fluorescent antibody (brand-CST, product number-#4410S) was added and incubated in the dark for 1 hour. After washing with PBS, the cells were resuspended in PBS and detected by flow cytometry.

[0168] [Added by reference (Rule 20.6) 24.02.2025] The test results are shown in Figure 2, which is a flow cytometry analysis of the binding peaks of monoclonal antibody K47-20C10-B8 with Klebsiella pneumoniae encapsulated bacteria HVKP4 and non-encapsulated bacteria 19-249ΔwbaP (capsular polysaccharide expression gene knockout), and Figure 3 is a flow cytometry analysis of the binding peaks of monoclonal antibody K47-18G6-B2 with Klebsiella pneumoniae HVKP4 and bacteria 19-249ΔwbaP (capsular polysaccharide expression gene knockout).

[0169] [Added by reference (Rule 20.6) 24.02.2025] As shown in Figure 2, monoclonal antibody K47-20C10-B8 can bind to the highly virulent strain HVKP4 of Klebsiella pneumoniae, but does not bind to Klebsiella pneumoniae 19-249ΔwbaP (capsular polysaccharide expression gene knockout). It can be seen that monoclonal antibody K47-20C10-B8 can be used in the diagnosis and typing of clinical strains.

[0170] [Added by reference (Rule 20.6) 24.02.2025] Referring to Figure 3 , monoclonal antibody K47-18G6-B2 was tested for binding to Klebsiella pneumoniae HVKP4. The results showed that monoclonal antibody K47-18G6-B2 could bind to encapsulated strains but not to Klebsiella pneumoniae 19-249ΔwbaP (capsular polysaccharide expression gene knockout). This indicates that monoclonal antibody K47-18G6-B2 can be used for the diagnosis and typing of clinical strains.

[0171] [Added by reference (Rule 20.6) 24.02.2025] Example 5: Bactericidal test of monoclonal antibodies against Klebsiella pneumoniae K47 capsular polysaccharide

[0172] [Added by reference (rule 20.6) 24.02.2025] This example shows the opsonophagocytic bactericidal test of the K47 type Klebsiella pneumoniae capsular polysaccharide monoclonal antibody provided by the present application, specifically: Carbapenem-resistant Klebsiella pneumoniae HVKP4 strain was diluted 10 5 CFU / mL, 10 μL / well was added to a 96-well cell working plate. The Klebsiella pneumoniae capsular polysaccharide monoclonal antibody provided in this application was diluted in a gradient manner and added to the above cell working plate at 20 μL / well. The bacteria and the antibody were incubated at 700 rpm / min for 30 minutes. The HL-60 cells differentiated by DMF were washed with HBSS buffer and adjusted to a concentration of 1×10 7 / mL, and then 1×10 7Mix cells (cells / mL) with diluted complement at a 1:4 volume ratio. Add 50 μL / well of the mixture to a 96-well plate. Place the 96-well plate on a mixer and incubate in a 5% CO2, 37°C CO2 incubator with shaking for 45 minutes. After opsonophagocytosis is stopped, spot the plate onto a blood agar plate and incubate overnight in a CO2 incubator. Calculate the bactericidal efficiency of each antibody dilution.

[0173] [Incorporated by reference (Rule 20.6) 24.02.2025] As shown in FIG4 , the in vitro bactericidal curves of K47-5B9-F8 and K47-15D7-B10, monoclonal antibodies against Klebsiella pneumoniae capsular polysaccharide provided by this application, against the HVKP4 strain of Klebsiella pneumoniae are shown. FIG4 shows that K47-5B9-F8 and K47-15D7-B10 have a good bactericidal effect against the highly virulent HVKP4 strain in vitro, with the highest bactericidal rates reaching 91.5% and 95.5%, respectively. This is significantly different from the negative control antibody.

[0174] [Incorporated by reference (Rule 20.6) 24.02.2025] Example 6: Prophylactic test of monoclonal antibodies to Klebsiella pneumoniae K47 capsular polysaccharide

[0175] [Added by reference (Rule 20.6) 24.02.2025] This example shows the protective test of the K47 Klebsiella pneumoniae capsular polysaccharide monoclonal antibody provided by the present application. Specifically, six Klebsiella pneumoniae capsular polysaccharide monoclonal antibodies were administered by intraperitoneal injection to five mice per group, with each mouse receiving 0.25 mg. PBS was used as a negative control. Four hours later, the K47 Klebsiella pneumoniae highly virulent strain HVKP4 was used at a concentration of 8.5×10 5 Balb / c mice (n=5) were infected intraperitoneally with the bacterial count of CFU. The survival rate of the mice was observed 10 days after infection, and a survival curve was drawn.

[0176] [Added by reference (Rule 20.6) 24.02.2025] As shown in FIG5 , which is the survival curve of the K47 type Klebsiella pneumoniae capsular polysaccharide monoclonal antibody preventive test provided by this application, it can be seen that after 4 hours of administration of the six K47 type Klebsiella pneumoniae capsular polysaccharide monoclonal antibodies and the control PBS, the amount of Klebsiella pneumoniae K47 type bacteria (strain number HVKP4) was 8.5×10 5 After CFU challenge, the survival rate of the control group was 20%, while the survival rates of the five monoclonal antibody groups were all 100%, with statistically significant differences (p < 0.0001).

[0177] [Added by reference (Rule 20.6) 24.02.2025] In summary, the above embodiments are merely preferred embodiments of the present application and are not intended to limit the scope of protection of the present application. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present application shall be included in the scope of protection of the present application.

Claims

1. A monoclonal antibody against Klebsiella pneumoniae capsular polysaccharide, characterized in that: The Klebsiella pneumoniae is K47 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 complementarity determining regions of the light chain variable region comprise three sequences as shown in SEQ ID:3, SEQ ID:4 and SEQ ID:5, 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:3, SEQ ID:4 and SEQ ID:5; the complementarity determining regions of the heavy chain variable region comprise three sequences as shown in SEQ ID:6, SEQ ID:7 and SEQ ID:8, 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:6, SEQ ID:7 and SEQ ID:

8.

3. A hybridoma cell line, characterized in that The hybridoma cell line is named K47-5B9-F8, and the deposit number of the hybridoma cell line is CCTCC NO: C2023314. The hybridoma cell line is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody according to claim 1.

4. A monoclonal antibody against Klebsiella pneumoniae capsular polysaccharide, characterized in that: The Klebsiella pneumoniae is K47 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 regions of the light chain variable region comprise 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 regions of the heavy chain variable region comprise 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 K47-15D7-B10, and the deposit number of the hybridoma cell line is CCTCC NO: C2023312. The hybridoma cell line is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody according to claim 4.

7. A monoclonal antibody against Klebsiella pneumoniae capsular polysaccharide, characterized in that: The Klebsiella pneumoniae is K47 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 regions of the light chain variable region comprise 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 regions of the heavy chain variable region comprise 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 K47-18G6-B2, the deposit number of the hybridoma cell line is CCTCC NO: C2023311, and the hybridoma cell line is used to prepare the Klebsiella pneumoniae pod polysaccharide monoclonal antibody according to claim 7.

10. A monoclonal antibody against Klebsiella pneumoniae capsular polysaccharide, characterized in that: The Klebsiella pneumoniae is K47 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 regions of the light chain variable region comprise 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 regions of the heavy chain variable region comprise 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 K47-6E2, and the deposit number of the hybridoma cell line is CCTCC NO: C2023296. The hybridoma cell line is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody according to claim 10.

13. A monoclonal antibody against Klebsiella pneumoniae capsular polysaccharide, characterized in that: The Klebsiella pneumoniae is K47 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 regions of the light chain variable region comprise three sequences as shown in SEQ ID:35, SEQ ID:36 and SEQ ID:37, 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:35, SEQ ID:36 and SEQ ID:37, and the complementarity determining regions of the heavy chain variable region comprise three sequences as shown in SEQ ID:38, SEQ ID:39 and SEQ ID:40, 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:38, SEQ ID:39 and SEQ ID:

40.

15. A hybridoma cell line, characterized in that The hybridoma cell line is named K47-20 C10-B8, and the deposit number of the hybridoma cell line is CCTCC NO: C2023313. The hybridoma cell line is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody according to claim 13.

16. A monoclonal antibody against Klebsiella pneumoniae capsular polysaccharide, characterized in that: The Klebsiella pneumoniae is K47 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 regions of the light chain variable region comprise 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 regions of the heavy chain variable region comprise 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 K47-4H 8, and the deposit number of the hybridoma cell line is CCTCC: C2024339. The hybridoma cell line is used to prepare the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody according to claim 16.

19. 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 and 16, and further comprises any one or more combinations of pharmaceutically acceptable carriers, excipients or diluents.

20. A kit for detecting Klebsiella pneumoniae K47, characterized in that: The kit for detecting Klebsiella pneumoniae type K47 comprises a monoclonal antibody against the Klebsiella pneumoniae capsular polysaccharide according to any one of claims 1, 4, 7, 10, 13 and 16, and further comprises a positive control, a negative control, an antibody diluent, a color developing solution, a stop solution, a blocking solution or a washing solution, in combination with any one or more thereof.

21. Use of the Klebsiella pneumoniae capsular polysaccharide monoclonal antibody according to any one of claims 1, 4, 7, 10, 13 and 16 in the preparation of a drug for treating K47 type Klebsiella pneumoniae infection and / or a detection product.

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