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Ciprofloxacin and clenbuterol bispecific fused antibody and application thereof

A fusion antibody, bispecific technology, applied in application, specific peptides, introduction of foreign genetic material using vectors, etc., can solve problems such as poor stability, limited ScFv popularity, and short half-life.

Active Publication Date: 2014-04-02
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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  • Ciprofloxacin and clenbuterol bispecific fused antibody and application thereof
  • Ciprofloxacin and clenbuterol bispecific fused antibody and application thereof
  • Ciprofloxacin and clenbuterol bispecific fused antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Preparation of Ciprofloxacin and Clenbuterol Single Chain Antibodies

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

Embodiment 2

[0022] Example 2 Expression of ciprofloxacin and clenbuterol single-chain antibody 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 constructed respectively. Taking the construction of pET-22b-CPFX-ScFv as an example, see details figure 1 For the construction of pET-22b-CL-ScFv plasmid, see 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. The target fragments of double-enzyme digestion were purified and recovered, connected according to the system in Table 2, and centrifuged briefly to mix at the bottom of the tube. Connect overnight at 4°C and set aside.

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

[0026]

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

[0028]

[0029] Table 3 Two kinds of single-chain ...

Embodiment 3

[0035] Example 3 Construction of fusion antibody

[0036] Three pairs of primers (synthesized by Shanghai Sangon) for amplifying the variable region genes of single-chain antibodies were designed and synthesized.

[0037] Table 4 Primers for amplifying each gene

[0038]

[0039] (“—” is the added protective base, the underlined part is the introduced enzyme cleavage site, the enzyme cleavage sites are ECOR I and Not I respectively, the italic black box is the added connecting peptide, and the bold is the added stop codon)

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

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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 an application thereof, 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 C Due to its huge structural characteristics, it has many drawbacks in disease diagnosis and treatment: first, the heterogeneity of traditional monoclonal antibodies makes the induction of immune responses generally stronger; second, it metabolizes quickly in the body and is slightly less specific; Most of the screened monoclonal antibodies can only maintain the activity of natural antibodies, and the selected antibodies have no ability to neutralize the biological activity of the antigen. It is quite complicated and difficult to select neutralizing antibodies. Finally, many specific antib...

Claims

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Application Information

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Patent Type & Authority Applications(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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