Anti-HSF5 monoclonal antibody and application thereof

Abstract

The invention belongs to the technical field of biology, and relates to an anti-HSF5 monoclonal antibody and application thereof, the monoclonal antibody can recognize HSF5 protein, the monoclonal antibody comprises a heavy chain variable region and a light chain variable region, the light chain variable region has three complementary determining regions of amino acid sequences as shown in SEQ ID NO: 1-SEQ ID NO: 3, and the light chain variable region has three complementary determining regions of amino acid sequences as shown in SEQ ID NO: 2-SEQ ID NO: 3. The heavy chain variable region has three complementary determining regions of amino acid sequences as shown in SEQ ID NO: 4-SEQ ID NO: 6. The anti-HSF5 monoclonal antibody provided by the invention has good recognition specificity on in-vitro purified human source HSF5 protein; compared with a polyclonal antibody, the monoclonal antibody has higher recognition specificity.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology and relates to an anti-HSF5 monoclonal antibody and application thereof. Background Art

[0002] Heat shock transcription factors (HSFs) are regulators of heat shock protein (HSPs) expression, mediate responses to various forms of physiological and environmental stimuli, and play an important role in differentiation and development. HSFs in the human genome include HSF1, HSF2, HSF4, HSF5, HSFX, and HSFY. Current research mainly focuses on HSF1 and HSF2 family members. So far, only a few studies have focused on HSF5. Saju JM et al. found that knocking out Hsf5 in zebrafish causes severe infertility in male zebrafish, but its function and mechanism of action in mammalian spermatogenesis are still unclear. Currently, the commercially available antibodies against human HSF5 protein are mainly rabbit polyclonal antibodies, such as HSF5 Polyclonal antibody (17765-1-AP, proteintech), but the commercially available antibodies have poor recognition specificity for human HSF5 protein. Therefore, it is necessary to develop a monoclonal antibody against HSF5. Summary of the invention

[0003] In order to solve the technical problem, the present invention provides an anti-HSF5 monoclonal antibody and its application, which can specifically recognize HSF5 protein, has good antibody specificity and high titer, and compared with polyclonal antibodies, the monoclonal antibody has stronger recognition specificity.

[0004] In one aspect, the present invention provides a monoclonal antibody against HSF5, wherein the monoclonal antibody can recognize HSF5 protein.

[0005] The monoclonal antibody comprises a heavy chain variable region and a light chain variable region:

[0006] The light chain variable region has three complementarity determining regions of the amino acid sequences shown in SEQ ID NO: 1 to SEQ ID NO: 3;

[0007] The heavy chain variable region has three complementarity determining regions of the amino acid sequences shown in SEQ ID NO: 4 to SEQ ID NO: 6.

[0008] Furthermore, the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 19, and the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 20.

[0009] Furthermore, the light chain framework sequence is shown in SEQ ID NOs: 11 to 14; the heavy chain framework sequence is shown in SEQ ID NOs: 15 to 18.

[0010] Furthermore, the monoclonal antibody also includes:

[0011] An antibody with the same function obtained by replacing, deleting and / or adding one or more amino acids to the amino acid sequence of the monoclonal antibody;

[0012] or comprising a heavy chain variable region having an amino acid sequence that is at least 80% homologous to the heavy chain variable region; and a light chain variable region having an amino acid sequence that is at least 80% homologous to the light chain variable region;

[0013] Or an antibody obtained by connecting a tag to the N-terminus and / or C-terminus of the monoclonal antibody.

[0014] In a second aspect, the present invention provides a nucleic acid molecule encoding the monoclonal antibody, wherein the nucleic acid molecule comprises a nucleic acid molecule encoding the heavy chain variable region and a nucleic acid molecule encoding the light chain variable region.

[0015] In a third aspect, the present invention provides an expression vector comprising the nucleic acid, wherein the expression vector is capable of expressing the nucleic acid in a prokaryotic or eukaryotic host cell.

[0016] In a fourth aspect, the present invention provides an engineered bacterium or eukaryotic host cell comprising the expression vector.

[0017] The fifth aspect of the present invention provides a use of the monoclonal antibody of HSF5 protein in preparing a HSF5 protein detection reagent or kit.

[0018] In a sixth aspect, the present invention provides a use of the monoclonal antibody of the HSF5 protein in preparing a quality control antibody for a HSF5 protein colloidal gold detection kit.

[0019] In a seventh aspect, the present invention provides a colloidal gold rapid detection test strip for HSF5 protein, comprising:

[0020] A bottom plate, a sample absorption pad, a conjugation pad, a chromatography matrix and a water absorption pad bonded to the bottom plate and overlapped in sequence; wherein the conjugation pad is coated with a colloidal gold complex coated with a monoclonal antibody of the HSF5 protein; a quality control line C is provided on the side of the chromatography matrix close to the conjugation pad, and a detection line T is provided on the side of the chromatography matrix close to the water absorption pad; the quality control line C is coated with an anti-mouse IgG secondary antibody; and the detection line T is coated with the monoclonal antibody of the HSF5 protein.

[0021] The present invention has the following advantages and beneficial effects:

[0022] The anti-HSF5 monoclonal antibody provided by the invention has good recognition specificity for both in vitro purified human HSF5 protein and exogenously expressed human HSF5 protein in cells; compared with polyclonal antibodies, the monoclonal antibody has stronger recognition specificity.

[0023] The HSF5 heavy chain antibody of the present invention forms a stable bond with the HSF5 purified protein, proving that the HSF5 heavy chain antibody has good affinity and specificity for the HSF5 purified protein. The HSF5 light chain antibody forms a stable bond with the HSF5 purified protein, proving that the HSF5 light chain antibody has good affinity and specificity for the HSF5 purified protein. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a diagram of the purification of the human HSF5 truncated protein used to immunize mice in Example 1;

[0025] Figure 2 This is a graph showing the results of the heavy chain antibody membrane interference experiment in Example 2, where the horizontal axis is time and the vertical axis is the thickness of the biosensor surface film layer; Note: Steps ① balance; ② solidification; ③ cleaning; ④ binding; ⑤ dissociation; ⑥ renaturation; ⑦ cleaning;

[0026] Figure 3 This is a graph showing the results of the light chain antibody membrane interference experiment in Example 2, where the horizontal axis is time and the vertical axis is the thickness of the biosensor surface film layer; Note: Steps ① balance; ② solidification; ③ cleaning; ④ binding; ⑤ dissociation; ⑥ renaturation; ⑦ cleaning;

[0027] Figure 4 The Western Blot experiment in Example 3 was used to identify the specific recognition between the anti-human HSF5 protein polyclonal antibody and monoclonal antibody and the HSF5 protein, respectively. The sample was purified human HSF5 truncated protein; the left figure shows the recognition of the sample by the anti-human HSF5 protein polyclonal antibody, and the right figure shows the recognition of the sample by the anti-human HSF5 protein monoclonal antibody, respectively. DETAILED DESCRIPTION

[0028] For a better understanding of the present invention, the following examples are provided to further illustrate the present invention, but the present invention is not limited to the following examples.

[0029] Throughout the specification, unless otherwise specifically stated, the terms used herein should be understood as meanings commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which the present invention belongs. In the event of a conflict, the present specification takes precedence.

[0030] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0031] The monoclonal antibodies of the present application and their preparation methods and application effects are described in detail below in conjunction with the examples and experimental data. The specific experimental conditions and methods not specified in the following examples are usually in accordance with conventional conditions such as those described in J. Sambrook et al., ed., Molecular Cloning Experiment Guide (3rd edition), Science Press, 1992; DL Spector et al., Cell Experiment Guide, Science Press, 2001, or the conditions recommended by the manufacturer.

[0032] Example 1. Monoclonal antibody and preparation method thereof

[0033] 1. Construction of recombinant protein vector and prokaryotic expression of protein

[0034] (1) The human HSF5 protein truncation (543-571 amino acid sequence) was selected and its CDS sequence was cloned into the Pet42b vector to obtain the recombinant vector Pet42b-human HSF5.

[0035] Using human cDNA as template, the primer pair sequences used are:

[0036] Forward Primer:

[0037] 5'-tttaagaaggagatatacatatgggcccggcgagcaaaccgag-3';

[0038] 5'-gcaaaggaggaggtggaagcggcccggcgagcaaaccgag-3';

[0039] Reverse Primer:

[0040] 5'-gcttccacctcctcctttgccctgctggctgttgct-3';

[0041] 5'-tggtggtggtggtgctcgagtttgccctgctggctgttgctgc-3';

[0042] The gene was cloned into the NdeI-XhoⅠ restriction site of Pet42b vector.

[0043] (2) Protein expression and purification:

[0044] ①Plasmid transformation:

[0045] The constructed human HSF5 prokaryotic expression plasmid was transformed into DE3 expression competent cells, and sterile glass beads were used to slide on the LB solid plate containing kanamycin resistance to evenly spread the bacterial solution, and then placed in a constant temperature incubator at 37°C overnight.

[0046] ②Select single clone bacteria & protein expression:

[0047] After a single clone of bacteria grows on the LB plate, pick the single clone and transfer it to 1L of liquid culture medium containing kanamycin and shake it. Shake the bacteria overnight at 37℃ until the OD reaches 0. 600 =0.4-0.8, add 400L isopropyl-β-D-thiogalactoside (IPTG) for induction, and induce expression at 16°C for 16 hours.

[0048] ③Collecting and splitting bacteria:

[0049] The induced bacterial solution was centrifuged at 8000 rpm for 10 min, the supernatant was discarded, the bacteria were resuspended in 60 ml lysis buffer, lysozyme and phenylmethylsulfonyl fluoride (PMSF) (1:100) were added, and ultrasonically disrupted on ice for 1 hour. Then centrifuged twice at 9900 rpm for 10 min each time, and the supernatant was retained.

[0050] ④ Combined with Ni NTA beads:

[0051] Pipette 3 ml Ni NTA beads and wash twice with lysis buffer. After washing, add to the supernatant and incubate at 4°C for 12 hours.

[0052] ⑤ Elution protein:

[0053] After centrifugation, remove the supernatant and wash the Ni NTA beads with washing buffer for 3 times, 10 minutes each time. Then, elute the protein with elution buffer for 10 minutes each time, for a total of 3 times.

[0054] ⑥Concentrated protein:

[0055] Concentrate the protein using a concentrator tube and replace the PBS buffer while concentrating.

[0056] ⑦ Protein gel running:

[0057] After protein purification, run the gel, such as Figure 1 shown.

[0058] ⑧Protein storage:

[0059] The concentrated protein was stored at -80°C.

[0060] (3) Protein purification related reagent formula

[0061] Lysis buffer: NaH2PO4: 50 mM, NaCl: 300 mM, Imidazole: 10 mM, pH = 8.0;

[0062] Washing buffer: NaH2PO4: 50mM, NaCl: 300mM, Imidazole: 50mM, pH = 8.0;

[0063] Elution buffer: NaH2PO4: 50 mM, NaCl: 300 mM, Imidazole: 250 mM, pH = 8.0.

[0064] 2. Animal immunization

[0065] Purified human HSF5 protein was used as an antigen to immunize mice. Three 6-8 week old Balb / C mice were selected. Freund's complete adjuvant was used for the first main injection, and Freund's incomplete adjuvant was used for subsequent booster injections. Both were fully mixed with an equal volume of antigen before injection. The immunization method used multiple abdominal injections. The immunization dose was 100 μg antigen / experimental mouse for the main injection and 50 μg antigen / experimental mouse for the booster injection.

[0066] The immunization cycle is shown in Table 1.

[0067] deal with Immunization time Immunization dose Primary immunization 2022-10-24 0.10mg / piece Strengthen immunity 2022-11-7 0.05mg / piece Strengthen immunity 2022-11-21 0.05mg / piece Strengthen immunity 2022-12-5 0.05mg / piece

[0068] Table 1

[0069] 3. Antibody light chain and heavy chain plasmid construction and sequencing

[0070] (1) Extraction of RNA

[0071] Kill the immunized mice by cervical dislocation, and grind the spleen. Add 1mL Trizol reagent to the grinding tissue solution, centrifuge at 12000rpm for 5min, and discard the precipitate. Add 200mL chloroform, shake and mix, and let stand at room temperature for 15min. Centrifuge at 12000rpm for 15min, draw the upper aqueous layer into a new EP tube, add 0.5mL isopropanol, mix, and let stand at room temperature for 10min. Centrifuge at 12000rpm for 10min, and discard the supernatant. Add 1mL 75% ethanol, shake and centrifuge at 7500rpm for 5min, discard the supernatant, dry at room temperature, and add 50μL RNase-free double distilled water. Store at -80℃ for later use.

[0072] (2) Reverse transcription to prepare cDNA

[0073] ① 8 μL RNA, heat at 65°C for 5 min, quickly cool on ice, and leave on ice for 2 min;

[0074] ② Heat the mixture from the previous step, 5×gDNA wiper Mix 2μL, at 42℃ for 2min;

[0075] ③The mixture from the previous step, 10×RT Mix, HiScript III Enzyme Mix 2μL, Oligo(dT) 20 VN 1 μL, RNase-free ddH2O 5 μL, heating at 25°C for 5 min, heating at 37°C for 45 min, heating at 85°C for 5 sec.

[0076] (3) PCR amplification of cDNA

[0077] Primers were designed based on information in the UCSC Genome Browser Gateway database.

[0078] Light chain forward primer: 5′-attctagagctagcgaattcatggattttcaggtgcagattttcagcttcctg-3′;

[0079] Light chain reverse primer: 5′-atccttgcggccgcggatccctaacactcattcctgttgaagctcttgacaat-3′;

[0080] Heavy chain forward primer: 5′-ttctagagctagcgaattcatggctgtcctggcactgctcctc-3′;

[0081] Heavy chain reverse primer: 5′-tccttcgcggccgcggatcctcatttacccggagtccgggagaag-3′;

[0082] The heavy chain and light chain cDNAs were amplified separately: cDNA 5 μL, upstream primer 2 μL, downstream primer 2 μL, 2×PhantaFlash Master Mix 20 μL, RNase-free ddH2O 11 μL; PCR reaction was performed according to the following reaction conditions: incubation at 98°C for 5 min, denaturation at 98°C for 25 s, annealing at 63°C for 20 s, extension at 72°C for 25 s, and extension at 72°C for 5 min after 35 cycles.

[0083] (4) Agarose gel electrophoresis and gel recovery

[0084] The PCR products were subjected to agarose gel electrophoresis, and the electrophoresis results were observed. The amplified products with molecular weights of 700-800 bp and 1400-1600 bp were recovered from the gel to obtain the light chain and heavy chain DNA.

[0085] (5) Connection

[0086] Ligate the light chain and heavy chain DNA into the EcoRI-BamHI restriction site of the PB513B vector: 5 μL of vector, 2 μL of DNA, and NovoRec R Plus recombinase 1μL, 5× reaction buffer 4μL, RNase-free ddH2O 8μL, heat at 50℃ for 30min.

[0087] (6) Ligation product transformation

[0088] The ligation product was transformed into DH5α expression competent cells, and the bacterial solution was evenly spread on an ampicillin-resistant LB solid plate using sterile glass beads, and incubated in a 37°C constant temperature incubator overnight.

[0089] (7) Bacteria selection & plasmid sequencing

[0090] After the colonies grow on the LB plate, the colonies were picked up and placed in 30 mL of liquid culture medium containing ampicillin antibiotics and shaken at 37°C for 6 hours, and the plasmids were extracted and sent for sequencing. The results are shown in Table 2. The protein sequence encoding the complete light chain (SEQ ID NO: 7) and the protein sequence encoding the complete heavy chain (SEQ ID NO: 8) are shown in Table 3. The nucleic acid molecule sequence encoding the complete light chain (SEQ ID NO: 9) and the nucleic acid molecule sequence encoding the complete heavy chain (SEQ ID NO: 10) are shown in Table 4.

[0091] Sequence Light chain CDR1 SASSSIRSSYLH (SEQ ID NO: 1) Light chain CDR2 GTSILAS (SEQ ID NO: 2) Light chain CDR3 QQWSGYPFT (SEQ ID NO: 3) Heavy chain CDR1 SYGVH (SEQ ID NO:4) Heavy chain CDR2 VIWSDGSTTYNSALKS (SEQ ID NO:5) Heavy chain CDR3 QKIYYGYDGYAMDY (SEQ ID NO: 6)

[0092] Table 2

[0093]

[0094]

[0095] Table 3

[0096]

[0097]

[0098]

[0099] Table 4

[0100] 4. Transfection & Cell Culture

[0101] The antibody light chain and heavy chain plasmids with the correct sequence were selected and transfected into U2OS cells respectively. The cells were cultured and the culture medium was collected. The light chain and heavy chain antibodies were in the culture medium.

[0102] 5. Membrane interference experiment (BLI) to detect antibody specificity (see Example 2 for details)

[0103] 6. Co-transfection & cell culture

[0104] The plasmids of light chain and heavy chain antibodies that specifically bind to the HSF5 protein were selected and co-transfected into U2OS cells. Stable cell lines were constructed through puromycin selection, and the culture medium was collected.

[0105] 7. Recover antibodies using Protein A / G magnetic beads

[0106] (1) Magnetic bead pretreatment

[0107] Mix the magnetic beads by inversion, take 50μL of magnetic beads to a 1.5mL EP tube, add 500μL of binding buffer to resuspend and rinse the magnetic beads, centrifuge briefly, place the EP tube on a magnetic rack and let it stand for 30s, discard the supernatant, repeat the rinsing step once, add 100μL of binding buffer to resuspend the magnetic beads for later use.

[0108] (2) Protein A / G magnetic beads bind to antibodies

[0109] Add 500 μL of antibody-containing culture medium to the magnetic beads, mix thoroughly and incubate in a vertical rotating mixer at room temperature for 30 minutes. After instant centrifugation, place the EP tube on a magnetic stand for 30 seconds and discard the supernatant. Add 500 μL of binding buffer to resuspend and rinse the magnetic beads. After instant centrifugation, place the EP tube on a magnetic stand for 30 seconds and discard the supernatant. Repeat the rinsing step 3 times.

[0110] (3) Elution of Antibodies

[0111] Place the EP tube on the magnetic rack and let it stand for 30 seconds, discard the supernatant, add 50 μl elution buffer to resuspend the magnetic beads, and let it stand at room temperature for 10 minutes; after instant centrifugation, place the EP tube on the magnetic rack and let it stand for 30 seconds, transfer the supernatant to a new EP tube, and add 50 μl neutralization buffer.

[0112] (4) Relevant reagent formula

[0113] Binding buffer: 50 mM Tris, 150 mM NaCl, 1 mM EDTA, 1% NP-40, pH 7.4;

[0114] Elution buffer: 0.15 M glycine, pH 3.1;

[0115] Neutralization buffer: 1 M Tris-HCl, pH 8.0.

[0116] 8. Antibody Concentration & Antibody Storage

[0117] Concentrate the antibody using a concentrator tube and replace the PBS buffer while concentrating. Store the concentrated antibody at -80℃.

[0118] Example 2: Detection of antibody specificity using membrane interferometry (BLI)

[0119] (1) Balance: Place the biosensor (Sensor A1-H1) (Sartorius: NTA Biosensors, Item no. 18-5101) were immersed in PBST buffer for 3 min;

[0120] (2) Solidification: Prepare HSF5 purified protein stabilization buffer and BSA solution containing 0.1% Tween 20 in advance, immerse the biosensor in the protein stabilization buffer or BSA solution for 6 min to allow the protein to bind to the surface of the biosensor (Sensor A1 group to Sensor G1 group: HSF5 purified protein stabilization buffer; Sensor H1 group: BSA solution);

[0121] (3) Cleaning: Immerse the biosensor in PBST buffer and clean it for 8 min;

[0122] (4) Binding: The biosensor bound to the protein to be tested was immersed in antibody solutions of different concentrations for 30 min to allow the antibody to fully bind to the protein (Sensor A1 group: 1 mg / mL HSF5 purified protein stabilizing buffer; Sensor B1 group: 0.5 mg / mL HSF5 purified protein stabilizing buffer; Sensor C1 group: 0.25 mg / mL HSF5 purified protein stabilizing buffer; Sensor D1 group: 0.125 mg / mL HSF5 purified protein stabilizing buffer; Sensor E1 group: 0.0625 mg / mL HSF5 purified protein stabilizing buffer; Sensor F1 group: 0.03125 mg / mL HSF5 purified protein stabilizing buffer; Sensor G1 group: 0.015625 mg / mL HSF5 purified protein stabilizing buffer; Sensor H1 group: 1 mg / mL BSA solution);

[0123] (5) Dissociation: The biosensor was immersed in PBST buffer and washed for 8 min;

[0124] (6) Renaturation: The biosensor bound to the protein to be tested is immersed in 500 mM Imidazole elution solution for dissociation, so that the protein falls off the biosensor surface;

[0125] (7) Cleaning: Immerse the biosensor in PBST buffer and clean it for 8 min;

[0126] (8) The kinetic constants of the target protein to be tested were obtained by real-time monitoring of the thickness of the biosensor biofilm layer during the experiment. The heavy chain antibody detection results are shown in Figure 2 , light chain antibody test results are shown in Figure 3 .

[0127] Figure 2 The figure shows the affinity diagram of HSF5 heavy chain antibody and HSF5 antigen detected by membrane interference experiment. Steps ①-⑦ are: equilibrium, solidification, washing, binding, dissociation, renaturation, washing. It can be seen from the figure that in steps ④-⑤, compared with the control group (Sensor H1 group), the thickness of the biosensor surface film layer in the experimental group (Sensor A1 group-Sensor G1 group) increased, indicating that the HSF5 heavy chain antibody formed a stable combination with the HSF5 purified protein, proving that the HSF5 heavy chain antibody has good affinity and specificity for the HSF5 purified protein.

[0128] Figure 3 The figure shows the affinity diagram of HSF5 light chain antibody and HSF5 antigen detected by membrane interference experiment. Steps ①-⑦ are: equilibrium, solidification, washing, binding, dissociation, renaturation, washing. It can be seen from the figure that in steps ④-⑤, compared with the control group (Sensor H1 group), the thickness of the biosensor surface film layer in the experimental group (Sensor A1 group-Sensor G1 group) increased, indicating that the HSF5 light chain antibody formed a stable combination with the HSF5 purified protein, proving that the HSF5 light chain antibody has good affinity and specificity for the HSF5 purified protein.

[0129] Example 3: Detection of Antibody Specificity Using Western Blot Experiment

[0130] The HSF5 truncated protein was purified in vitro from prokaryotes, and the recognition specificity of the monoclonal antibody was tested using Western Blot experiments. Figure 4 As shown in the figure below, the left and right figures are Western Blot experiments to detect the specificity of anti-human HSF5 protein polyclonal antibody and anti-human HSF5 protein monoclonal antibody. The results show that both anti-human HSF5 protein polyclonal antibody and anti-human HSF5 protein monoclonal antibody can recognize HSF5 protein, but anti-human HSF5 protein monoclonal antibody has better recognition specificity for HSF5.

[0131] The above is only a preferred embodiment of the present invention, which certainly cannot be used to limit the scope of rights of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and changes can be made without departing from the principle of the present invention, and these improvements and changes are also regarded as the protection scope of the present invention.

Claims

1. A monoclonal antibody against HSF5, characterized in that: The monoclonal antibody can recognize HSF5 protein, The monoclonal antibody comprises a heavy chain variable region and a light chain variable region: The light chain variable region has three complementarity determining regions of the amino acid sequences shown in SEQ ID NO: 1 to SEQ ID NO: 3; The heavy chain variable region has three complementarity determining regions of the amino acid sequences shown in SEQ ID NO: 4 to SEQ ID NO:

6.

2. The anti-HSF5 monoclonal antibody according to claim 1, characterized in that The amino acid sequence of the light chain variable region is shown in SEQ ID NO: 19, and the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO:

20.

3. The anti-HSF5 monoclonal antibody according to claim 1, characterized in that The light chain framework sequence is shown in SEQ ID NOs: 11 to 14; the heavy chain framework sequence is shown in SEQ ID NOs: 15 to 18.

4. The anti-HSF5 monoclonal antibody according to claim 1, characterized in that The monoclonal antibody also includes: An antibody with the same function obtained by replacing, deleting and / or adding one or more amino acids to the amino acid sequence of the monoclonal antibody; or comprising a heavy chain variable region having an amino acid sequence that is at least 80% homologous to the heavy chain variable region; and a light chain variable region having an amino acid sequence that is at least 80% homologous to the light chain variable region; Or an antibody obtained by connecting a tag to the N-terminus and / or C-terminus of the monoclonal antibody.

5. A nucleic acid molecule encoding the anti-HSF5 monoclonal antibody according to any one of claims 1 to 4, characterized in that: The nucleic acid molecules include a nucleic acid molecule encoding the heavy chain variable region and a nucleic acid molecule encoding the light chain variable region.

6. An expression vector comprising the nucleic acid molecule of claim 5, characterized in that: The expression vector is capable of expressing the nucleic acid in a prokaryotic or eukaryotic host cell.

7. An engineered bacterium or eukaryotic host cell comprising the expression vector according to claim 6.

8. Use of the monoclonal antibody against HSF5 protein according to any one of claims 1 to 4 in the preparation of a reagent or kit for detecting HSF5 protein.

9. Use of the monoclonal antibody against HSF5 protein according to any one of claims 1 to 4 in preparing a quality control antibody for a HSF5 protein colloidal gold detection kit.

10. A colloidal gold rapid detection test strip for HSF5 protein, characterized in that: include: A base plate, a sample absorption pad, a conjugation pad, a chromatography matrix and a water absorbent pad bonded to the base plate and overlapped in sequence; wherein the conjugation pad is coated with a colloidal gold complex coated with an anti-HSF5 monoclonal antibody as described in any one of claims 1-4; a quality control line C is provided on the side of the chromatography matrix close to the conjugation pad, and a detection line T is provided on the side of the chromatography matrix close to the water absorbent pad; the quality control line C is coated with an anti-mouse IgG secondary antibody; the detection line T is coated with an anti-HSF5 monoclonal antibody as described in any one of claims 1-4.

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