Monoclonal antibody capable of specifically identifying HPV18 L1 protein and application of monoclonal antibody
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A monoclonal antibody and specific technology, applied in the field of HPV infection detection, can solve the problems of expensive equipment, inability to judge recovery and prognosis, and not suitable for large-scale screening of cervical cancer, and achieve high specificity and convenient detection
Active Publication Date: 2018-07-13
Y CLONE MEDICAL SCI CO LTD
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The equipment required by these two methods is relatively expensive, and the diagnosis of different genotypes of HPV is cumbersome, and it is not suitable for large-scale screening of cervical cancer
Commercial hybrid capture kits can detect HPV DNA in clinical samples without PCR amplification, and can distinguish between high-risk and low-risk types, but such kits cannot identify specific genotypes infected with HPV
In addition, the test results can only show the existence of the HPV virus gene, but cannot detect the state of the HPV virus, whether it is actively replicating or dead virus, and cannot judge whether it is in recovery or prognosis
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[0031] The principles and features of the present invention are described below in conjunction with examples, which are only used to explain the present invention and are not intended to limit the scope of the present invention.
[0032] 1. Immunogen design and monoclonal antibody preparation
[0033] We synthesized a modified polypeptide (L1M186) containing the specific epitope QSPVP from HPV18 L1 protein; and L1M186-coupled hemocyanin (KLH) constituted L1M186-KLH as an immunogen.
[0034]The immunogen was dissolved in phosphate buffered saline (PBS, pH7.4) at a concentration of 1 mg / ml. The primary immunization was performed at week 0, and each mouse was fully emulsified with 100 μl of 1 mg / ml immunogen solution and 100 μl of complete Freund’s adjuvant to form a water-in-oil emulsion, and immunized by multi-point abdominal subcutaneous injection. 6 mice. In the 2nd week, 5th week, 8th week, and 11th week, the booster immunization was carried out 4 times, and each time each...
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Abstract
The invention relates to a monoclonal antibody capable of specifically identifying HPV18 L1 protein. The monoclonal antibody comprises a heavy-chain region and a light-chain region, wherein the sequence of the heavy-chain region is SEQ ID NO: 1 or SEQ ID NO: 3, and a sequence which has 80 percent consistency with the SEQ ID NO: 1 or the SEQ ID NO: 3; the sequence of the light-chain region is SEQ ID NO: 2 or SEQ ID NO: 4, and a sequence which has 80 percent consistency with the SEQ ID NO: 1 or the SEQ ID NO: 3; the invention further relates to application of the monoclonal antibody to preparation of an HPV18 detection reagent, and further relates to an HPV18 detection reagent kit comprising the monoclonal antibody. The monoclonal antibody and the reagent and the kit, which are prepared fromthe monoclonal antibody, can be used for detecting patients in an HPV (Human Papillomavirus) active infection period; the detection is convenient and the specificity is high.
Description
technical field [0001] The invention relates to the field of detection of HPV infection, and more particularly relates to a monoclonal antibody capable of specifically recognizing HPV18L1 protein and its application, and a diagnostic kit prepared therefrom. Background technique [0002] Human papillomavirus (HPV) is a widely circulated sexually transmitted pathogen that causes genital warts. HPV is a papillomavacuovirus A genus belonging to the Papovaviridae family, a spherical DNA virus, and an epitheliophilic virus with a high degree of specificity, which can cause squamous epithelial proliferation of human skin and mucous membranes. There are many subtypes of HPV, and more than 130 types have been isolated so far. Different types cause different clinical manifestations. According to the clinical consequences of infection, HPV is divided into high-risk types (HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 73, 82, etc.) and low-risk types (such as HPV6, 11, 40, 42...
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