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A kind of bispecific fusion antibody of ciprofloxacin and clenbuterol and its application

A technology of fusion antibody and ciprofloxacin, applied in application, specific peptide, introduction of foreign genetic material using carrier, etc., can solve the problems of poor stability, limited popularity of ScFv, short half-life, etc.

Active Publication Date: 2016-06-29
HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the further research on ScFv, while significant progress has been made, some problems in clinical application have also been exposed, such as low affinity, poor stability, short half-life and other shortcomings, which also limit the use of ScFv in the treatment process. popularity in

Method used

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  • A kind of bispecific fusion antibody of ciprofloxacin and clenbuterol and its application
  • A kind of bispecific fusion antibody of ciprofloxacin and clenbuterol and its application
  • A kind of bispecific fusion antibody of ciprofloxacin and clenbuterol and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The preparation of embodiment 1 ciprofloxacin and clenbuterol single-chain antibody

[0021] Ciprofloxacin and clenbuterol single-chain antibodies were obtained by chemical synthesis. The nucleotide sequences encoding the ciprofloxacin single-chain antibody and the clenbuterol single-chain antibody are respectively shown in SEQ ID NO.2 and SEQ ID NO.3.

Embodiment 2

[0022] Example 2 Ciprofloxacin and clenbuterol single-chain antibody expressed in pET-22b (+) prokaryotic vector

[0023] Using the two genes in Example 1 and the pET-22b expression system, pET-22b-CPFX-ScFv and pET-22b-CL-ScFv were respectively constructed. Take the construction of pET-22b-CPFX–ScFv as an example, see figure 1 , The construction of pET-22b-CL-ScFv plasmid refers to the construction method of pET-22b-CPFX-ScFv.

[0024] The double enzyme digestion system of the target gene product and the expression vector plasmid is shown in Table 1. Purify and recover the double-digested target fragment, connect according to the system in Table 2, and centrifuge briefly to mix at the bottom of the tube. Incubate overnight at 4°C and set aside.

[0025] Table 1 pET-22b-CPFX-ScFv double enzyme digestion system

[0026]

[0027] Table 2 pET-22b-CL-ScFv double enzyme digestion system

[0028]

[0029] Table 3 Two kinds of scFv double enzyme digestion linking system ...

Embodiment 3

[0035] Example 3 Construction of Fusion Antibody

[0036] Three pairs of primers (synthesized by Shanghai Sangon) for amplifying the variable region gene of the single-chain antibody were designed and synthesized. The sequence of each primer, the starting position in the whole genome and the added enzyme cutting site are shown in Table 3.

[0037] Table 4 Amplifies the primers of each gene

[0038]

[0039] ("—" is the added protective base, the underlined part is the introduced restriction site, the restriction site is ECORⅠ and NotⅠ respectively, the italicized black box is the added connecting peptide, and the bolded is the added stop codon)

[0040]Analyze the gene sequence characteristics of the single-chain antibody series, design and synthesize 2 pairs of primers, each pair of primers introduces a second connecting peptide different from the assembled single-chain antibody gene, and obtain two recombinant bacteria prepared by PCR method. The fusion gene fragment of ...

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Abstract

The invention discloses a ciprofloxacin and clenbuterol bispecific fused antibody and application thereof and belongs to the technical field of immune rapid screening. Genes VH and VL of a ciprofloxacin single-chain antibody and a clenbuterol single-chain antibody are integrated to obtain a multi-specific antibody capable of generating reaction with two residual medicaments. A rapid immune detection method is constructed by using the multi-specific fused single-chain antibody as a probe, so that the aim of rapidly screening various residual medicaments at a time is fulfilled.

Description

technical field [0001] The invention relates to a bispecific fusion antibody of ciprofloxacin and clenbuterol and its application, in particular to an antibody capable of immunologically reacting with two residual drugs at the same time, belonging to the technical field of rapid immune screening. Background technique [0002] Traditional monoclonal antibodies due to their molecular F C The huge structural characteristics make it have many disadvantages in the diagnosis and treatment of diseases: firstly, the heterogeneity of traditional monoclonal antibodies makes the immune response generally strong; secondly, the metabolism is fast in the body and the specificity is slightly poor; thirdly, the hybridoma technology Most of the monoclonal antibodies screened can only maintain the activity of natural antibodies, and the selected antibodies have no ability to neutralize the biological activity of antigens. It is quite complicated and difficult to select neutralizing antibodies...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/44C12N15/13C12N15/70C12N1/21G01N33/53
Inventor 王颖张东杰柳增善姚笛霍方珍
Owner HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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