Feline parvovirus recombinant protein and preparation of monoclonal antibody of recombinant protein

Abstract

The invention belongs to the technical field of bioengineering, and relates to a feline parvovirus recombinant protein which comprises two dominant epitopes of a FPV (feline parvovirus) antigen. In order to improve the yield of the recombinant protein in a prokaryotic expression system, the amino acid sequence of the recombinant protein is converted into the corresponding nucleotide sequence by the aid of Escherichia coli biased codons, the nucleotide sequence is chemically synthesized, and a recombinant expression vector is constructed. The invention further relates to a preparation method ofa monoclonal antibody of the feline parvovirus recombinant protein. According to the preparation method, immunization, cell fusion and multiple screening are implemented to obtain a hybridoma cell strain, the monoclonal antibody is purified, colloidal gold particles are marked, an optimal monoclonal antibody matched composition is determined through an orthogonal experiment, and the recombinant protein can be used for early diagnosis of feline parvovirus infection.

Description

technical field [0001] The invention belongs to the technical field of bioengineering. Specifically, the present invention relates to a new feline parvovirus recombinant protein, the nucleotide sequence of the feline parvovirus recombinant protein is coded and synthesized by genetic engineering technology, a plasmid vector containing the above nucleotide sequence is constructed, and the above plasmid is transformed The Escherichia coli strain of the vector expresses feline parvovirus recombinant protein; the recombinant protein is used to prepare a monoclonal antibody, and is applied to the preliminary diagnosis of feline parvovirus infection. Background technique [0002] Feline parvovirus (Feline parvovirus, FPV) is also known as feline panleukopenia virus, feline infectious enteritis virus, and feline distemper virus. The clinical manifestations of cats infected with parvovirus are mainly high fever, vomiting, diarrhea and enteritis, which can lead to a decrease in the nu...

AI Technical Summary

Problems solved by technology

Mainly there is the isolation and identification of feline parvovirus. This method uses cultured cells to observe cell lesions, but some viruses proliferate slowly and cannot grow in cell culture. In the process of transportation and transportation, the infectivity may be lost, and this method requires the operator to have certain experience, and the cost is relatively high

The sensitivity of the electron microscopy test is low, and different viruses with similar shapes cannot be distinguished, and the sample processing volume is large, which limits the application of this method

RT-PCR method is used to detect feline parvovirus nucleic acid. This method has good specificity and sensitivity, can quickly distinguish viruses, and is suitable for simultaneous detection of a large number of samples. However, the equipment is expensive, the requirements for operators are high, the detection time is long, and the detection cost is high.

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