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Lincosamides universal monoclonal antibody hybridoma cell strain and application thereof

A monoclonal antibody and monoclonal technology, applied in biochemical equipment and methods, instruments, microorganisms, etc., can solve problems such as expensive instruments, complicated operations, and long time

Active Publication Date: 2018-06-15
JIANGNAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the instrument detection method is accurate, but the instrument is expensive, the operation is complicated, the time is long, and the sample pretreatment is cumbersome.

Method used

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  • Lincosamides universal monoclonal antibody hybridoma cell strain and application thereof
  • Lincosamides universal monoclonal antibody hybridoma cell strain and application thereof
  • Lincosamides universal monoclonal antibody hybridoma cell strain and application thereof

Examples

Experimental program
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Embodiment 1

[0031] Example 1: Preparation of lincosamide-type universal monoclonal antibodies

[0032] (1) Design of hapten: It is prepared by the condensation reaction of β-mercaptopropionic acid and clindamycin. See the specific synthetic route figure 1 . Dissolve 9.6g of clindamycin in 20mL dimethylsulfoxide (DMSO), add 2.3g KOH, and slowly add β-mercaptopropionic acid solution (3.2g β-mercaptopropionic acid dissolved in 10mL DMSO) while stirring. The oil bath was heated to 100°C and reacted for 2h. After the product was naturally cooled to room temperature, 50mL of water was added, and the reaction solution was adjusted to pH 3 with 6mol / L HCl, and then extracted with 50mL of dichloromethane. The extract was dried over anhydrous Na2SO4, concentrated Crystallized to obtain yellow crystals, namely the target compound, which was identified by mass spectrometry.

[0033] (2) Preparation of complete antigen: using clindamycin chloride substitution derivatives, coupled with bovine serum albumin...

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Abstract

The invention relates to a lincosamides universal monoclonal antibody hybridoma cell strain and an application thereof, which belong to the technical field of food safety immunology detection. The lincosamides universal monoclonal antibody hybridoma cell strain employs a lincosamide substituted derivative as semiantigen, semiantigen is coupled with bovine serum albumin (BSA) through an activated ester method to obtain immunization antigen, the immunization antigen and a freund's adjuvant are uniformly mixed, a mixture is used for immunization BALB / c mice through hypodermic injection, clindamycin is coupled with albumin from chicken egg white (OVA) by employing a carbonyl diimidazole (CDI) method, and the material is used for detecting mice serum and cell supernatant as coating antigen, thespleen cell of the immunization mice is subjected to fusion with mice myeloma cells through a PEG method, and through indirect ELISA and indirect competition ELISA screening and three-time subclone,the group selected hybridoma cell strain can be obtained. The cell has good inhibition effect for clindamycin, lincomycin and pirlimycin, and can satisfy the requirement on lincosamides stumpy immunodetection products on the market.

Description

Technical field [0001] The invention relates to a lincosamide universal monoclonal antibody hybridoma cell strain and its application, and belongs to the technical field of food safety immunological detection. Background technique [0002] Lincosamides have the activity against Gram-positive aerobes and Gram-positive or negative anaerobes. The commonly used veterinary clinics are lincomycin, clindamycin and pyriamycin. [0003] Lincosamide drug residues in animal-derived foods can cause renal dysfunction and increased drug resistance of gram-positive bacteria. Both my country and the European Union have made relevant regulations on the maximum residue limits in animal-derived foods. Therefore, it is urgent to establish a Fast, sensitive and accurate detection methods for lincosamide drug residues ensure the safety of animal foods. [0004] The current methods for detecting Lincosamide drug residues mainly include high performance liquid chromatography (HPLC), liquid mass spectrometry...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/44G01N33/577C12R1/91
CPCG01N33/577C07K16/44G01N2430/00G01N33/9446C07K2317/92
Inventor 匡华郭玲玲胥传来徐丽广马伟刘丽强宋珊珊吴晓玲
Owner JIANGNAN UNIV
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