CN108276491B - Monoclonal antibody capable of specifically recognizing HPV18L1 protein and application thereof - Google Patents
Monoclonal antibody capable of specifically recognizing HPV18L1 protein and application thereof Download PDFInfo
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Abstract
The invention relates to a monoclonal antibody capable of specifically recognizing HPV18L1 protein, which comprises a heavy chain region and a light chain region, wherein the sequence of the heavy chain region is SEQ ID NO 1 or 3 or a sequence with 80% of identity with SEQ ID NO 1 or 3; the sequence of the light chain region is SEQ ID NO 2 or 4, or a sequence with 80% identity to SEQ ID NO 1 or 3; also relates to the application of the monoclonal antibody in preparing HPV18 detection reagent; also relates to an HPV18 detection kit containing the monoclonal antibody. By using the monoclonal antibody, the reagent and the kit prepared by the monoclonal antibody, a patient in the active infection stage of HPV can be detected, the detection is convenient, and the specificity is high.
Description
Technical Field
The invention relates to the field of HPV infection detection, in particular to a monoclonal antibody capable of specifically recognizing HPV18L1 protein, application thereof and a diagnostic kit prepared from the same.
Background
Human Papilloma Virus (HPV) is a widespread sexually transmitted pathogen that causes condylomata. HPV is a papilloma vacuolato virus A belonging to papovaviridae, a spherical DNA virus, is an epitheliophilic virus, has high specificity, and can cause squamous epithelial proliferation of human skin mucosa. HPV subtypes are very numerous, and over 130 species have been isolated so far, with different types causing different clinical manifestations. According to the clinical consequences of infection, HPV is classified into high-risk types (HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 73, 82, etc.) and low-risk types (HPV 6,11,40,42-44,54,61,70,72,81, etc.). Most HPV infections can be cured within 1-2 years, and the low-risk HPV types mainly cause exogenous condyloma lesions, flat condyloma lesions and low-grade Cervical Intraepithelial Neoplasia (CINI) of the perianal skin of the genital tract and the lower part of the vagina, are transient and can be naturally reversed. The high-risk HPV mainly causes cervical intraepithelial neoplasia (CINI-III level) and cervical cancer, and the CINI level of continuous high-risk HPV infection is easy to progress to CINI II-III level, which causes female cervical cancer. Cervical cancer is the second most serious cancer of women, and the clinical application proves that 95 to 99.7 percent of cervical cancer cases are related to high-risk HPV virus infection, namely HPV infection is a prerequisite trigger for cervical cancer. Scientific research finds that at least 20 high-risk types of HPV are related to 95% of early-stage invasive and invasive cervical cancer, particularly two subtypes of HPV16 and HPV18 are main causes of cervical cancer, and more than 70% of cervical cancer is related to the two subtypes.
Cervical cancer is one of the most common malignant tumors threatening the health of women worldwide, and the infection rate of HPV in the female population is around 13.5%. About 52.8 ten thousand new cases of cervical cancer exist in the world every year, the number of deaths related to the cervical cancer reaches 26.6 ten thousand, and research data in 2017 show that about 13 ten thousand new cases of cervical cancer exist in China every year, and the number of deaths is 3 ten thousand. In HPV DNA typing of 253 different cervical lesions by using Polymerase Chain Reaction (PCR), the HPV positive rates are in a gradually increasing trend according to the sequence of chronic cervicitis, pseudocondyloma, wart-like lesions, condyloma acuminatum, CIN and cervical cancer, and are respectively 16.7%, 29.8%, 37.5%, 61.7%, 79.7% and 90.9%. After HPV16DNA and HPV18DNA in 44 cervical cancer tissues, 18 paracarcinoma tissues, 30 chronic cervicitis and 15 normal cervical tissues are detected by using an In Situ PCR (ISPCR) technology, the HPV16DNA in the 44 cervical cancer tissues is positive 79.5 percent, and the HPV18DNA is positive 11.3 percent; 15 cases of paracancerous tissues with grade CINI-III, HPV16DNA positive 80%, and 15 cases of normal cervical tissues HPV DNA negative. These results show that the precancerous lesion and cervical cancer are closely related to high-risk HPV16/18 infection, especially HPV16/18 infection, and the detection rate of HPV16/18 is increased with the progressive lesion degree, which shows that the detection of HPV16/18 is an important index for predicting cervical cancer. However, in most cases internationally, the detection of high-risk HPV is only carried out when there is an apparent lesion in the cervix, and the optimal intervention and treatment opportunity is missed. Therefore, the early HPV typing detection is necessary to detect the high-risk HPV subtype (especially HPV 16/18).
Typing detection of HPV16/18 may provide the following benefits: 1) can provide diagnosis early warning in the early stage of virus infection; 2) can be used as an evaluation index of the treatment effect of doctors, shows the infection of a single type of high-risk subtype by HPV typing detection twice continuously, shows that the possibility of cervical cancer occurrence is increased, and is expected to be paid great attention; 3) HPV infection is different in different regions, the predominant types are different, and typing detection is favorable for research and vaccine prevention and control of HPV infection in different regions.
Commonly used HPV detection methods are liquid-based thin-layer cell detection (TCT) and HPV nucleic acid detection. TCT is mainly diagnosed by observing the morphological characteristics of cells, and the method is noninvasive and simple, so that the TCT can be clinically used as a primary screen in cervical cancer screening and has a certain guiding effect on subsequent examination and treatment. However, TCT detection is subjective and the stage of HPV infection and the subtype of HPV infection cannot be known with certainty.
HPV nucleic acid detection can determine the infected HPV subtypes, and is generally realized by a real-time fluorescent PCR method or a hybridization capture method. The two methods both require expensive equipment, are complicated to diagnose and operate for different genotypes of HPV, and are not suitable for large-scale screening of cervical cancer. Commercial hybridization capture method kit can detect HPV DNA in clinical samples without PCR amplification, and can distinguish high-risk type and low-risk type, but the kit can not identify specific genotype of infecting HPV. In addition, the detection result only indicates the existence of HPV viral genes, the existence state of the HPV virus, whether the virus is actively replicated or killed, and whether the virus is recovered or not and the prognosis cannot be judged.
Therefore, a new HPV infection detection kit is needed.
Disclosure of Invention
The HPV L1 protein is the viral capsid protein of HPV, and different HPV subtypes encode different capsid proteins, and the existence of which can reflect that the HPV virus is in an active replication stage. The research result shows that the positive expression rate of the high-risk HPV L1 capsid protein shows a descending trend along with the aggravation of cervical lesion degree. The clinical detection of positive high-risk HPV L1 capsid protein indicates that the body is infected by high-risk HPV and the virus is in the replication stage. Therefore, the HPV infection subtype and infection status can be detected by specifically detecting the HPV L1 capsid protein. In the research process, the inventor selects the unique amino acid sequence of HPV18 to synthesize antigen, and induces a high specificity monoclonal antibody aiming at HPV18L1 protein.
Based on the above research, the invention provides a monoclonal antibody capable of specifically recognizing HPV18L1 protein, which is characterized by comprising a heavy chain region and a light chain region, wherein the sequence of the heavy chain region is SEQ ID NO. 1 or 3, or a sequence with 80% identity to SEQ ID NO. 1 or 3; the sequence of the light chain region is SEQ ID NO 2 or 4, or a sequence with 80% identity with SEQ ID NO 1 or 3.
In a preferred embodiment, SEQ ID NO 1 has three CDR regions, respectively, segments 26-33, 52-58 and 97-106.
In a preferred embodiment, there are three CDR regions in SEQ ID NO. 2, the (27 th to 37 th segments, the 55 th to 57 th segments and the 95 th to 103 th segments, respectively.
In one embodiment, the sequence of the heavy chain region is selected from SEQ ID NOs 1, 3, 5-7.
In one embodiment, the sequence of the light chain region is selected from SEQ ID NOs 2, 4, 8-20.
In a preferred embodiment, the heavy chain region sequence of the monoclonal antibody is SEQ ID NO 1 and the light chain sequence is SEQ ID NO 19.
In a preferred embodiment, the heavy chain region sequence of the monoclonal antibody is SEQ ID NO 6 and the light chain sequence is SEQ ID NO 2.
The invention also discloses application of the monoclonal antibody in preparation of an HPV18 detection reagent.
The invention also discloses a kit for detecting HPV18 infection, which comprises the monoclonal antibody.
Preferably, the kit is an ELISA kit comprising a capture antibody which is the monoclonal antibody of any one of claims 1-7 and a detection antibody which is a polyclonal antibody prepared using HPV18L1 coat protein as an antigen.
The invention also discloses a method for detecting HPV18, which comprises the step of detecting pathological samples from patients by using the monoclonal antibody or the reagent or the kit prepared by the monoclonal antibody.
By using the monoclonal antibody, the reagent and the kit prepared by the monoclonal antibody, a patient in the active infection stage of HPV can be detected, the detection is convenient, and the specificity is high.
Drawings
FIG. 1 is a statistical graph of the ELISA detection of serum titers of mouse sera against the immunogen naked peptide L1M186 after different immunizations;
FIG. 2 is a statistical chart of ELISA for detecting the affinity of 4 monoclonal cell supernatants for L1M186 polypeptide;
FIG. 3 is a statistical chart of the affinity of the 10C9 or 10D8 antibodies for different polypeptides in which the QSPVP amino acid is mutated to alanine (A) one by one at a fixed point based on the QSPVP amino acid in the L1M186 polypeptide, the underlined amino acid is the amino acid at the corresponding position of QSPVP;
FIG. 4 is a statistical plot of the LISA detection kit for determining antibody subtypes of 10C9 and 10D 8;
FIG. 5 is a statistical plot of the affinity of 16 mutant clones for HPV18L1 coat protein;
FIG. 6 is a graph comparing the protein sequences of wild type and 15 mutant clones with higher affinity to HPV18L1 coat protein;
FIG. 7 photograph of fluorescent staining of clinical samples of different HPV subtypes with monoclonal antibody;
FIG. 8 is a statistical chart of HPV18 ELISA detection kit for detecting different subtype HPV infectors;
FIG. 9 is a statistical curve of the sensitivity of the HPV18 ELISA detection kit for detecting HPV18L1 protein;
FIG. 10 is a statistical curve of the sensitivity of HPV18 ELISA detection kit for detecting HPV18L1 particles.
Detailed Description
The principles and features of this invention are described below in conjunction with examples, which are set forth to illustrate, but are not to be construed to limit the scope of the invention.
1. Immunogen design and monoclonal antibody preparation
We synthesized a modified polypeptide containing the specific epitope QSPVP from HPV18L1 protein (L1M 186); and coupling L1M186 to hemocyanin (KLH) to form L1M186-KLH as an immunogen.
The immunogen was dissolved in phosphate buffered saline (PBS, pH7.4) at a concentration of 1 mg/ml. At week 0, the mice were primed by 100. mu.l of 1mg/ml immunogen solution per mouse and 100. mu.l of Freund's complete adjuvant, thoroughly emulsified to form a water-in-oil emulsion, and immunized by multiple abdominal subcutaneous injections to immunize 6 mice. 4 booster immunizations are carried out at 2 weeks, 5 weeks, 8 weeks and 11 weeks, respectively, wherein each mouse is immunized with 50 ul of 1mg/ml polypeptide solution and 50 ul of emulsion of Freund's incomplete adjuvant; blood was collected at 6 weeks, 9 weeks, and 12 weeks.
Fusing mouse myeloma cells (Sp2/0) and spleen cells of an immunized mouse by a conventional PEG method within 1 week after the last immunization, cloning cells by a limiting dilution method, screening specific antibodies by indirect ELISA, continuously subcloning positive cells for 2 times, and preserving the cloned cell strain in liquid nitrogen after the cell strain is determined to be a stable cell strain through passage. The obtained monoclonal cell strain was injected into the abdominal cavity to prepare ascites.
The ELISA results of mouse serum antibodies are shown in figure 1, with the mean titer of 6 mice after 3 immunizations being 10600-fold dilution; the mean titer of serum from 6 mice after 4 immunizations was 46900-fold dilution, and the mean titer of serum from 6 mice after 5 immunizations was 72500-fold dilution. The above experimental results show that the serum titer presents an increasing trend with the increase of the number of immunizations, indicating that the immunization with L1M186-KLH is successful and that specific antibodies with high affinity are induced. After 5 immunization, the serum titer of the No. 5 mouse (marked by a triangle) is highest, and the dilution is 102400 times; therefore, the 5# mouse was selected to be subjected to cell fusion and subcloning after the shock immunization, and four monoclonal cells 1B3, 10a8, 10C9 and 10D8 were obtained.
The results of ELISA testing on monoclonal cell supernatants are shown in FIG. 2, and compared with cell supernatants of 1B3 and 10A8, cell supernatants of 10C9 and 10D8 showed better binding with the immunogenic polypeptide L1M186, and both read values were greater than 2.0; meanwhile, 10C9 and 10D8 do not react with HPV16 subtype polypeptide L1M161 and other polypeptide L1M185 and L1M188 of HPV 18. These results indicate that cell supernatants of 10C9 and 10D8 specifically recognized the L1M186 polypeptide of HPV18 without cross-reactivity with other unrelated polypeptides.
2. Monoclonal antibody identification
2.1 epitope identification
QSPVP 5 amino acids in the L1M186 polypeptide are coated with different polypeptides subjected to alanine (A) site-directed mutagenesis one by one to carry out ELISA detection. As shown in fig. 3, the binding value of the antibodies 10C9 and 10D8 to the original L1M186 polypeptide was 1.6, and when the aspartic acid (Q) at the first position, the proline (P) at the third position, or the valine (V) at the fourth position of the L1M186 polypeptide was mutated to alanine (a), the binding value of ELISA was reduced to blank control level, which indicates that the three amino acids Q, P, V play a crucial role in the binding of the 10C9 and 10D8 to the L1M186 polypeptide and are important epitopes for antibody binding. In addition, when the second serine (S) and the fifth proline (P) in the L1M186 polypeptide are mutated into alanine (A), the ELISA binding value is also reduced by half, thereby indicating that the two amino acids play important roles in the binding of 10C9 and 10D8 with the L1M186 polypeptide, but are not as critical as QPV. The above experimental results indicate that the antibody binding epitope of 10C9 and 10D8 is QSPVP, where QPV plays a crucial role. The detection of the ELISA typing detection kit shows that 10C9 and 10D8 are IgG1 subtype monoclonal antibodies (FIG. 4).
2.2 monoclonal antibody affinity identification
To examine the affinity of 10C9 and 10D8 to L1 protein, we utilized surface plasmon resonanceThe technology detects that the affinity of 10C9 and 10D8 to L1-HPV18 protein is 6.6x10 respectively-8And 3.5x10-9The results show that both 10C9 and 10D8 have strong affinity for HPV18L1 protein.
2.3 sequencing of monoclonal antibodies
After obtaining monoclonal cell strains of 10C9 and 10D8, RNA is extracted from the cell strains, and is simultaneously inverted into cDNA, an antibody scFv gene is amplified and sequenced, and after codon translation, a VH sequence SEQ ID NO 1 and a VL sequence SEQ ID NO 2 of 10C9 are obtained; and the VH sequence SEQ ID NO 3 and the VL sequence SEQ ID NO 4 of 10D 8. Wherein the VH sequence of 10C9 has three CDR regions, segment 26-33, segment 52-58 and segment 97-106; the VL sequence of 10C9 has three CDR regions, segments 27-37, 55-57, and 95-103.
The CDR regions of 10C9 were subjected to amino acid saturation mutagenesis and the corresponding 10C9 affinity matured phage antibody library was constructed. ELISA detection results are shown in FIG. 5, 48 phage monoclonals in a phage library of 10C9 mutant enriched with L1-HPV18 protein are selected for ELISA verification with L1-HPV18 protein, and the experimental results are shown in FIG. 5, and 16 clones (10C9-041 to 10C9-056) show binding with L1-HPV18 protein. The antibody sequence of 10C9-044 is completely consistent with that of wild-type 10C9, the ELISA binding value of the monoclonal phage is 0.35, and the experiment of the L1-HPV18 protein enrichment phage library is successful. It is noted that the binding values of the two monoclonal phages 10C9053 and 10C9055 to the L1-HPV18 protein are 1.25 and 0.73 respectively, and are significantly improved compared with the binding value of the wild-type 10C9-044 phage, which suggests that the two mutant forms of 10C9 may significantly improve the affinity of 10C9 to the L1-HPV18 protein.
The sequence pairs of the 16 clones are shown in FIG. 6, and among the 16 clones, the VH sequences of 10C9-054, 10C9-055 and 10C9-056 are shown in SEQ ID NO:5-7, and the VH sequences of the remaining clones are shown in SEQ ID NO: 1. The VL sequences of 10C9-041, 10C9-042, 10C9-043, 10C9-044, 10C9-045, 10C9-046, 10C9-047, 10C9-048, 10C9-049, 10C9-050, 10C9-051, 10C9-052, 10C9-053 and 10C9-056 are shown in SEQ ID NO 8-21, and the VL sequences of the rest clones are shown in SEQ ID NO 2.
2.4 characterization of monoclonal antibody specificity
To verify whether 10C9 specifically binds to HPV18, we tested 10C9 binding to different patient samples by immunofluorescence. Immunofluorescence results as shown in fig. 7, 10C9 specifically recognized cervical tissue samples from HPV18 patients and showed strong green positive fluorescence signals, but no green positive fluorescence signals were seen in cervical tissue sample staining of HPV16, HPV44 and healthy persons.
HPV18 infection ELISA detection kit
One or more of the monoclonal antibodies are used as capture antibodies to prepare detection antibodies capable of specifically recognizing HPV capsid protein or HPV virus (preferably HPV18 capsid protein or HPV18 virus), and other related detection reagents known in the art are prepared to form the HPV18 infection ELISA detection kit.
We used HPV18L1 protein to immunize New Zealand white rabbits to prepare polyclonal antibodies against the HPV18L1 coat protein. The specific method comprises the following steps: at week 0, the primary immunization was performed by thoroughly emulsifying 500. mu.l of 0.2mg/ml protein solution with 500. mu.l Freund's complete adjuvant to form water-in-oil emulsion, and performing multiple subcutaneous dorsal injections to immunize 2 New Zealand white rabbits. 4 times of booster immunization at 2 weeks, 5 weeks, 8 weeks and 11 weeks respectively, wherein each immunization is carried out on 500 mu l x 0.2mg/ml polypeptide solution and 500 mu l emulsion of Freund's incomplete adjuvant; blood sampling detection is carried out at 6 weeks, 9 weeks and 12 weeks.
ELISA results show that the two New Zealand white rabbits immunized by the L1 protein can induce the immunity of up to 1.9x10 after being immunized by 57The high-titer antiserum with the dilution multiple indicates that the self-made L1 protein has good immunogenicity, and the polyclonal serum is used as a detection antibody in the subsequent detection reagent development after being purified and is named as PAb-18L 1.
After condition exploration and optimization, an ELISA kit is constructed by using 10C9 as a capture antibody and PAb-18L1 as a detection antibody. By patient sample testing, we found that our ELISA test kit did not recognize patient samples of other HPV subtypes (like HPV16, HPV35, HPV39, HPV53, HPV 56, HPV58, HPV66), but specifically recognized HPV18 (FIG. 8). Therefore, the ELISA immune rapid diagnosis kit prepared by the inventor is a patient sample for specifically detecting HPV 18. In addition, the results of the experiment show that the sensitivity of the ELISA detection kit prepared by the inventor to HPV18L1 protein is 0.2ng (FIG. 9), and the sensitivity to HPV18 pseudovirus is 2.9ng (FIG. 10).
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Sequence listing
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Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Thr
85 90 95
Thr His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg
<210> 9
<211> 113
<212> PRT
<213> Artificial sequence
<400> 9
Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly
1 5 10 15
Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser
20 25 30
Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Gly Gln Thr
85 90 95
Thr His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg
<210> 10
<211> 113
<212> PRT
<213> Artificial sequence
<400> 10
Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly
1 5 10 15
Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser
20 25 30
Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Lys Leu Leu Ile Tyr Asp Val Ser Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Thr
85 90 95
Thr His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg
<210> 11
<211> 113
<212> PRT
<213> Artificial sequence
<400> 11
Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly
1 5 10 15
Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser
20 25 30
Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Thr
85 90 95
Thr His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg
<210> 12
<211> 113
<212> PRT
<213> Artificial sequence
<400> 12
Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly
1 5 10 15
Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser
20 25 30
Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Thr
85 90 95
Thr His Val Ser Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg
<210> 13
<211> 113
<212> PRT
<213> Artificial sequence
<400> 13
Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly
1 5 10 15
Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Leu
20 25 30
Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Thr
85 90 95
Thr His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg
<210> 14
<211> 113
<212> PRT
<213> Artificial sequence
<400> 14
Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly
1 5 10 15
Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Arg
20 25 30
Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Thr
85 90 95
Thr His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg
<210> 15
<211> 113
<212> PRT
<213> Artificial sequence
<400> 15
Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly
1 5 10 15
Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser
20 25 30
Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asn Leu Gly Val Tyr Phe Cys Asn Gln Thr
85 90 95
Thr His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg
<210> 16
<211> 113
<212> PRT
<213> Artificial sequence
<400> 16
Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly
1 5 10 15
Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser
20 25 30
Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Lys Leu Leu Ile Tyr Lys Val Ile Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asn Leu Gly Val Tyr Phe Cys Ser Gln Thr
85 90 95
Thr His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg
<210> 17
<211> 110
<212> PRT
<213> Artificial sequence
<400> 17
Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly
1 5 10 15
Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser
20 25 30
Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Lys Leu Leu Ile Tyr Lys Val Cys Asn Arg Phe Ser Gly Pro Gly
50 55 60
Phe Tyr Leu Leu Pro Phe Phe Ser Val Ser Gly Ser Leu Arg His Leu
65 70 75 80
Pro Ala Leu Val Pro Ala Glu Thr Gly Ser Val Ser Glu Thr Ala Asp
85 90 95
Leu Leu Cys Phe Pro Phe Leu Trp Cys Ser Gly Pro Phe Xaa
100 105 110
<210> 18
<211> 113
<212> PRT
<213> Artificial sequence
<400> 18
Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly
1 5 10 15
Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Val
20 25 30
Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asn Leu Gly Val Tyr Phe Cys Ser Gln Thr
85 90 95
Thr His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg
<210> 19
<211> 114
<212> PRT
<213> Artificial sequence
<400> 19
Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly
1 5 10 15
Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser
20 25 30
Asn Gly Asn Phe Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Lys Leu Leu Ile Tyr Lys Val Cys Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asn Leu Gly Val Tyr Phe Cys Ser Gln Thr
85 90 95
Thr Leu Val Pro Pro Thr Phe Arg Trp Trp Tyr Gln Thr Gly Asn Gln
100 105 110
Thr Xaa
<210> 20
<211> 113
<212> PRT
<213> Artificial sequence
<400> 20
Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly
1 5 10 15
Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser
20 25 30
Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asn Leu Gly Val Tyr Phe Cys Ser Gln Thr
85 90 95
Thr His Phe Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg
<210> 21
<211> 114
<212> PRT
<213> Artificial sequence
<400> 21
Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly
1 5 10 15
Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser
20 25 30
Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Thr
85 90 95
Thr His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg Xaa
Claims (4)
1. A monoclonal antibody capable of specifically recognizing HPV18L1 protein, which is characterized by comprising a heavy chain region and a light chain region, wherein the variable region sequence of the heavy chain region is SEQ ID NO 1; the variable region sequence of the light chain region is one of SEQ ID NO 2, 8-10 and 12-20.
2. The application of the monoclonal antibody in preparing the HPV18 detection reagent comprises the steps that the sequence of the variable region of the heavy chain region of the monoclonal antibody is shown as SEQ ID NO. 1, and the sequence of the variable region of the light chain region is shown as SEQ ID NO. 2.
3. A kit for detecting HPV18, which is characterized by comprising a monoclonal antibody, wherein the sequence of the variable region of the heavy chain region of the monoclonal antibody is shown as SEQ ID NO. 1, and the sequence of the variable region of the light chain region is shown as SEQ ID NO. 2.
4. The kit according to claim 3, wherein the kit is an ELISA kit comprising a capture antibody and a detection antibody, the capture antibody is a monoclonal antibody having a heavy chain region variable region sequence shown in SEQ ID NO. 1 and a light chain region variable region sequence shown in SEQ ID NO. 2, and the detection antibody is a polyclonal antibody prepared by using HPV18L1 capsid protein as an antigen.
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| CN201810110881.0A CN108276491B (en) | 2018-02-05 | 2018-02-05 | Monoclonal antibody capable of specifically recognizing HPV18L1 protein and application thereof |
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| CN110964104B (en) * | 2019-12-25 | 2021-08-13 | 南京薇熙生物医药科技有限公司 | Protein capable of binding HPV18 virus and application thereof |
| CN111793136A (en) * | 2020-05-11 | 2020-10-20 | 廊坊天光生物技术有限公司 | NMP22 antibody pair and application thereof |
| CN114230659B (en) * | 2021-11-12 | 2023-05-23 | 郑州大学 | anti-HPV 53L1 protein monoclonal antibody, preparation and application thereof |
| CN115112885B (en) * | 2022-06-18 | 2023-07-28 | 杭州爱光健康科技有限公司 | HPV detection kit and preparation method and application thereof |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1996029413A2 (en) * | 1995-03-22 | 1996-09-26 | Merck & Co., Inc. | Dna encoding human papilloma virus type 18 |
| US20090104215A1 (en) * | 2005-09-13 | 2009-04-23 | Irena Ekiel | Methods and compositions for modulating tumor cell activity |
| CN105175537A (en) * | 2012-06-08 | 2015-12-23 | 厦门大学 | Broad-spectrum monoclonal antibody resisting HPV L1 protein or antigen binding fragment thereof and application of broad-spectrum monoclonal antibody and antigen binding fragment |
| CN105524166A (en) * | 2014-09-28 | 2016-04-27 | 艾托金生物医药（苏州）有限公司 | Monoclonal antibody for identifying HPV18 positive cervical epithelial cancer cells, and applications thereof |
| CN105669859A (en) * | 2014-11-12 | 2016-06-15 | 北京康乐卫士生物技术股份有限公司 | Human papilloma virus 18 monoclonal antibody and application thereof |
| CN105753980A (en) * | 2016-03-29 | 2016-07-13 | 上海市普陀区中心医院 | HPV18 E6 monoclonal antibody and preparation method and application thereof |
| US20160368974A1 (en) * | 2013-12-12 | 2016-12-22 | Umc Utrecht Holding B.V. | Immunoglobulin-like molecules directed against fibronectin-eda |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5874089A (en) * | 1995-10-02 | 1999-02-23 | Georgetown University School Of Medicine | Protecting against canine oral papillomavirus (copy) |
-
2018
- 2018-02-05 CN CN201810110881.0A patent/CN108276491B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1996029413A2 (en) * | 1995-03-22 | 1996-09-26 | Merck & Co., Inc. | Dna encoding human papilloma virus type 18 |
| US20090104215A1 (en) * | 2005-09-13 | 2009-04-23 | Irena Ekiel | Methods and compositions for modulating tumor cell activity |
| CN105175537A (en) * | 2012-06-08 | 2015-12-23 | 厦门大学 | Broad-spectrum monoclonal antibody resisting HPV L1 protein or antigen binding fragment thereof and application of broad-spectrum monoclonal antibody and antigen binding fragment |
| US20160368974A1 (en) * | 2013-12-12 | 2016-12-22 | Umc Utrecht Holding B.V. | Immunoglobulin-like molecules directed against fibronectin-eda |
| CN105524166A (en) * | 2014-09-28 | 2016-04-27 | 艾托金生物医药（苏州）有限公司 | Monoclonal antibody for identifying HPV18 positive cervical epithelial cancer cells, and applications thereof |
| CN105669859A (en) * | 2014-11-12 | 2016-06-15 | 北京康乐卫士生物技术股份有限公司 | Human papilloma virus 18 monoclonal antibody and application thereof |
| CN105753980A (en) * | 2016-03-29 | 2016-07-13 | 上海市普陀区中心医院 | HPV18 E6 monoclonal antibody and preparation method and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| Epitope design of L1 protein for vaccine production against Human Papilloma Virus types 16 and 18;Sunanda Baidya等;《Bioinformation》;20170331;第13卷(第3期);第86-93页 * |
| HPV18L1基因重组DNA构建及其免疫小鼠体内特异抗体的测定;贾士东等;《中国免疫学杂志》;20000725;第16卷;第356-358页 * |
| Rational Design of Peptide Vaccines Against Multiple Types of Human Papillomavirus;Sumanta Dey等;《Cancer Informatics》;20160601;第15卷;第1-16页 * |
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