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Agglutinative Moraxella catarrhalis monoclonal antibody as well as preparation method and application thereof

A technology of Moraxella monoclonal antibody, which is applied in the field of biomedicine, can solve the problems of complex operation steps, high cost, and long detection time, achieve sensitive detection, strictly prevent missed detection and false detection, and improve sensitivity and detection rate Effect

Active Publication Date: 2020-05-15
GUANGZHOU WONDFO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003]Currently, the detection methods for Moraxella catarrhalis infection mainly include serological detection, pathogen culture detection and nucleic acid detection, but all of them have long detection time and operational difficulties. Due to the disadvantages of complicated steps and high cost, it is of great significance to prepare highly specific and highly targeted monoclonal antibodies for the rapid, simple and accurate detection of Moraxella catarrhalis

Method used

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  • Agglutinative Moraxella catarrhalis monoclonal antibody as well as preparation method and application thereof
  • Agglutinative Moraxella catarrhalis monoclonal antibody as well as preparation method and application thereof
  • Agglutinative Moraxella catarrhalis monoclonal antibody as well as preparation method and application thereof

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Embodiment 1

[0047] 1. Materials and methods

[0048] 1.1 Test materials and instruments

[0049] 1.1.1 Materials

[0050] The bacterial strain used in the specific embodiment is as follows:

[0051]M.catarrhalis(ATCC25238, ATCC8176), Streptococcus pneumoniae S.pneumoniae(ATCC25238), Mycoplasma pneumoniae M.Pneumonia(ATCC15531), Haemophilus influenzae H.influenzae(ATCC9007), Legionella pneumophila L.pneumophila(ATCC33152)

[0052] 1.1.2 Experimental animals

[0053] Female Balb / c mice: 6-8 weeks old.

[0054] 1.1.3 Main reagents and consumables

[0055] RPM1640, fetal bovine serum (FBS), HAT, HT, penicillin and streptomycin solution and polyethylene glycol (PEG, Mv4000) were purchased from Gibco, USA; SP2 / 0 myeloma cells; complete Freund's adjuvant, incomplete Freund's adjuvant, bovine serum albumin (BSA), Tween-20, and cyclophosphamide were all purchased from Sigma Company in the United States; the enzyme-labeled secondary antibody kit for mouse monoclonal antibody Ig class / subclass ...

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Abstract

The invention relates to a preparation method of an agglutinative Moraxella catarrhalis monoclonal antibody, which comprises the following steps: (1) obtaining non-agglutinative Moraxella catarrhalis,carrying out conventional culture to obtain a bacterial solution, carrying out ultrasonic disruption on the bacterial solution in an ice bath, and collecting the supernatant to obtain a tolerant; (2)obtaining agglutinative Moraxella catarrhalis, carrying out conventional culture to obtain a bacterial solution, carrying out ultrasonic crushing on the bacterial solution in an ice bath, and collecting supernatant to obtain immunogen; (3) tolerance stage: carrying out primary immunization on the mouse by adopting the tolerance antigen, respectively injecting cyclophosphamide (Cy) into the abdominal cavities of the mouse within 10 + / -2 minutes, 24 + / -1 hours and 48 + / -1 hours, and measuring titer; and (4) an immunization stage: after the mouse is tolerated, continuing to immunize the mouse byadopting the immunogen and the immunologic adjuvant, determining titer, and carrying out hybridoma cell fusion to obtain the agglutinative Moraxella catarrhalis monoclonal antibody. The antibody is high in purity, strong in affinity and high in specificity.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the agglutinating Moraxella catarrhalis monoclonal antibody and its preparation method and application. Background technique [0002] Moraxella catarrhalis (MC) is a Gram-negative diplococcus first discovered in 1896, when it was called Micrococcus catarrhalis, which originally belonged to the genus Neisseria, and was later renamed Branham catarrhalis. For a long time, everyone believed that MC was a normal resident bacterium of the human upper respiratory tract. Until the early 1980s, its role as a pathogen became more and more obvious. Specific clinical conditions of this organism, such as otitis media and chronic lung disease, are seen with increasing frequency in patients and have been implicated as causative agents of several infections, including sinusitis, conjunctivitis and laryngitis. In addition, sporadic cases of meningitis, endocarditis, and sepsis have been reported. The...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/531G01N33/558
CPCG01N33/577G01N33/531G01N33/558
Inventor 康业王羽吴培钿何小维黄幼珍
Owner GUANGZHOU WONDFO BIOTECH
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