Expression and Purification of the scFv from Hybridoma Cells Secreting a Monoclonal Antibody Against S Protein of PEDV

Zhu, Qinghe; Guo, Donghua; Feng, Li; Sun, Dongbo

doi:10.1089/mab.2012.0089

Cited by 12 publications

(9 citation statements)

References 27 publications

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“…Artificial passive immunization by oral administration of specific antibodies represents an attractive approach against gastrointestinal pathogens such as PEDV [26]. Several PEDV neutralizing antibodies were developed previously [15,16,32,33], and they might be good candidates for use as PEDV prophylactic agents. However, an effective antibody production system with low production cost is required for the animal industry.…”

Section: Discussionmentioning

confidence: 99%

Rapid Transient Production of a Monoclonal Antibody Neutralizing the Porcine Epidemic Diarrhea Virus (PEDV) in Nicotiana benthamiana and Lactuca sativa

et al. 2017

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Porcine epidemic diarrhea virus (PEDV) causes acute diarrhea, vomiting, dehydration, weight loss, and high mortality rate in neonatal piglets. Porcine epidemic diarrhea (PED) has been reported in Europe, America, and Asia including Thailand. The disease causes substantial losses to the swine industry in many countries. Presently, there is no effective PEDV vaccine available. In this study, we developed a plant-produced monoclonal antibody (mAb) 2C10 as a prophylactic candidate to prevent the PEDV infection. Recently, plant expression systems have gained interest as an alternative for the production of antibodies because of many advantages, such as low production cost, lack of human and animal pathogen, large scalability, etc. The 2C10 mAb was transiently expressed in and lettuce using geminiviral vector. After purification by protein A affinity chromatography, the antibody was tested for the binding and neutralizing activity against PEDV. Our result showed that the plant produced 2C10 mAb can bind to the virus and also inhibit PEDV infection. These results show excellent potential for a plant-expressed 2C10 as a PEDV prophylaxis and a diagnostic for PEDV infection.

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Section: Discussionmentioning

confidence: 99%

Rapid Transient Production of a Monoclonal Antibody Neutralizing the Porcine Epidemic Diarrhea Virus (PEDV) in Nicotiana benthamiana and Lactuca sativa

et al. 2017

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“…At present, many small molecules are reported as diagnostic reagents, including monoclonal antibody, singlechain fragment variable (scFv), aptamer, and peptide ligand. (18)(19)(20)(21) Of those diagnostic reagents, monoclonal antibody is still the most effective diagnostic tool in the biomedical field.…”

Section: Discussionmentioning

confidence: 99%

Preparation of Monoclonal Antibodies Against Bovine Haptoglobin

Wang

Gu²,

Guo³

et al. 2014

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

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Female, 8-week-old BALB/c mice were immunized with purified recombinant proteins of the predicted immunodominant region of bovine haptoglobin (pirBoHp). Two monoclonal antibodies (MAbs), named 1B3 and 6D6, were prepared by conventional B lymphocyte hybridoma technique. Titers of ascitic fluid and cell culture supernatant of MAb 1B3 were 1:9.6 × 10(8) and 1:8.2 × 10(4), respectively, and that of MAb 6D6 were 1:4.4 × 10(5) and 1:1.0 × 10(4), respectively. The subtype of MAbs 1B3 and 6D6 was IgG1κ. In Western blot analysis, MAbs 1B3 and 6D6 could recognize the α-chain of native BoHp from plasma of dairy cows. These data indicated that MAbs 1B3 and 6D6 have a potential use for developing diagnostic reagents of BoHp.

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“…Moreover, the recombinant pirBoHp protein was purified according to the method described by Zhu and colleagues. (19)…”

Section: Expression and Purification Of Pirbohp Genementioning

confidence: 99%

A Polyclonal Antibody Against Recombinant Bovine Haptoglobin Expressed inEscherichia coli

Guo

Zhang²,

Li³

et al. 2013

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

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The nucleotide sequence of the predicted immunodominant region of bovine haptoglobin (pirBoHp), without the signal peptide sequence, was synthesized based on the codon usage bias of Escherichia coli. The synthesized pirBoHp gene was cloned into the prokaryotic expression vector pET-32a (+), which contains a His-tag. The recombinant pirBoHp protein was successfully expressed in E. coli BL21 (DE3) cells. Western blot analysis showed that the purified recombinant pirBoHp protein could be recognized by an anti-His-tag monoclonal antibody. Further investigations indicated that a polyclonal antibody against the recombinant pirBoHp protein could recognize the α and β chains of native bovine haptoglobin in a pooled plasma sample from dairy cattle suffering from foot rot.

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Expression and Purification of the scFv from Hybridoma Cells Secreting a Monoclonal Antibody Against S Protein of PEDV

Cited by 12 publications

References 27 publications

Rapid Transient Production of a Monoclonal Antibody Neutralizing the Porcine Epidemic Diarrhea Virus (PEDV) in Nicotiana benthamiana and Lactuca sativa

Rapid Transient Production of a Monoclonal Antibody Neutralizing the Porcine Epidemic Diarrhea Virus (PEDV) in Nicotiana benthamiana and Lactuca sativa

Preparation of Monoclonal Antibodies Against Bovine Haptoglobin

A Polyclonal Antibody Against Recombinant Bovine Haptoglobin Expressed inEscherichia coli

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