Anti-mannan-binding lectin monoclonal antibodies and kit

A monoclonal antibody and mannose technology, which is applied in the direction of anti-animal/human immunoglobulin, biological testing, and microbial-based methods, can solve the problems of unreliable detection results, insufficient sensitivity, and narrow detection range. Accurate and reliable detection results, wide detection range and high sensitivity

Inactive Publication Date: 2014-03-05
杨旸
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current kits for detecting the content of mannose-binding clusterin (MBL) in human serum or plasma have poor specificity, insufficient sensitivity, and narrow detection range, resulting in unreliable detection results.

Method used

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  • Anti-mannan-binding lectin monoclonal antibodies and kit
  • Anti-mannan-binding lectin monoclonal antibodies and kit
  • Anti-mannan-binding lectin monoclonal antibodies and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0122] Establishment of anti-MBL hybridoma cells

[0123] Immunization: Female BALB / c mice aged 8-12 weeks were routinely immunized with 20ug MBL (purified from healthy Chinese plasma) in the tripod and subcutaneously, repeated immunization after 3-4 weeks, and 2 weeks later Tail vein booster once.

[0124] Fusion: Take splenocytes from the mice 3 days after the last booster immunization, mix them with myeloma cells (provided by the Cell Department of China National Institute of Pharmaceutical and Biological Products) at a ratio of 5:1, and fuse with 50% PEG solution. In HAT medium at 37°C 5% CO 2 to cultivate.

[0125] Detection and selective culture of hybridoma cells: After 3-4 days of culture, replace half of the medium with HAT medium until the cells grow to 1 / 3-1 / 2 wells, then detect the cells, coat the antigen MBL, and take the hybridoma supernatant ( Primary antibody) was incubated with the antigen, washed to remove unbound antibody, enzyme-labeled goat anti-mouse s...

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Abstract

Provided are anti-mannan-binding lectin monoclonal antibodies and a kit. Hybridoma cell strains comprise hybridoma cell strains used for coating the anti-mannan-binding lectin monoclonal antibodies, and with the culture number being CGMCC No. 0001, and also comprise hybridoma cell strains used for labeling the anti-mannan-binding lectin monoclonal antibodies, and the culture number being CGMCC No. 0002. The hybridoma cell strains used for coating the anti-mannan-binding lectin monoclonal antibodies and the hybridoma cell strains used for labeling the anti-mannan-binding lectin monoclonal antibodies have different binding epitopes. The invention aims to provide anti-mannan-binding lectin monoclonal antibodies and a kit with strong specificity, high sensitivity, wide detection range and accurate and reliable detection results.

Description

technical field [0001] The invention relates to an anti-mannose-binding clusterin monoclonal antibody and a kit. Background technique [0002] Mannan (poly) sugar-binding lectin, English name: mannan-binding lectin, abbreviated as MBL. Mannose-binding lectin is a serum C-type lectin produced by the liver. It is an acute-phase protein that can bind to mannose. It can regulate pathogens with mannose, activate the complement system, participate in innate immune responses, activate complement, Regulates inflammation, promotes opsonophagocytosis, and clears apoptotic cells. When the adaptive immune system is immature (infancy) or has been suppressed (e.g. after organ transplantation, or during cancer chemotherapy) often the absence of MBL may lead to increased susceptibility to infection. MBL deficiency is associated with autoimmune diseases such as systemic lupus erythematosus, rheumatoid arthritis, and cystic fibrosis with poor prognosis. MBL constitutes the first line of de...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/18G01N33/68C12R1/91
Inventor 杨明非张庶民
Owner 杨旸
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