Canine parvovirus single-domain antibody, and preparation method and application thereof

A technology of canine parvovirus and single-domain antibody, which is applied in the direction of antibodies, antiviral agents, antiviral immunoglobulin, etc., can solve the problems of difficult to stimulate T cell immune response, complex molecular structure of whole antibody, low VHH immunogenicity, etc. problem, to achieve the effect of shortening the treatment cycle, high affinity and small molecular weight
CN103665152AInactive Publication Date: 2014-03-26SHANXI AGRI UNIV
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
SHANXI AGRI UNIV
Publication Date
2014-03-26
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses a canine parvovirus single-domain antibody, and a preparation method and an application thereof. The canine parvovirus single-domain antibody has a nucleotide sequence as shown in SEQIDNO.1. The preparation method comprises the steps: immunizing an alpaca by a canine parvovirus inactivated vaccine, and taking peripheral blood to obtain alpaca anti-canine parvovirus serum; extracting RNA in the peripheral blood, and carrying out reverse transcription to generate cDNA; utilizing specific primers for amplification, and purifying a VHH fragment; connecting the VHH fragment with a phagemid vector, transforming TG1, and constructing a canine parvovirus single-domain antibody library; and enriching the antibody library by utilizing a canine parvovirus structural protein VP2, introducing into an expression bacterial strain E.coil HB2151, and utilizing IPTG to induce the canine parvovirus single-domain antibody VHH having the biological activity to be expressed. The prepared canine parvovirus single-domain antibody has the detection activity of 15 [mu]g / ml and the detection sensitivity of 250 ng / ml, and can be used as drugs applied for treating dogs with canine parvovirus disease.
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Description

technical field

[0001] The invention belongs to the technical field of clinical veterinary medicine and antibody engineering, and relates to a virus antibody, in particular to a canine parvovirus single-domain antibody mainly relying on the heavy chain antibody of alpaca to realize its biological expression, and a preparation method of the antibody and apply. Background technique

[0002] Canine parvovirus disease is caused by canine parvovirus (Canine parvovirus, CPV). It is characterized by high morbidity, high mortality, and strong infectivity. It can occur all year round. Hemorrhagic enteritis type and myocarditis type canine parvovirus disease The mortality rates are 10% to 50% and 60% to 100%, respectively, which can cause serious economic losses. Vaccines to prevent canine parvovirus mainly include inactivated vaccines, attenuated vaccines and multiple vaccines, and optimized immunization procedures can achieve better immune effects. However, testing and investigati...

Claims

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