A Polyclonal Antibody Against Recombinant Bovine Haptoglobin Expressed inEscherichia coli

Abstract

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.

Preparation of Monoclonal Antibodies Against Bovine Haptoglobin

“…In a previous study, polyclonal antibody against the recombinant pirBoHp protein could recognize aand bchains of native bovine haptoglobin. (9) In the current study, the MAbs 1B3 and 6D6 only showed specific reaction with a-chain of the native BoHp. This result suggests that in the BoHp molecule, the a-chain may have better antigenicity than the b-chain does.…”

“…The recombinant protein of the predicted immunodominant region of bovine haptoglobin (pirBoHp) was expressed in a previous study. (9) The purified pirBoHp recombinant protein was stored at the Department of Veterinary Clinical Medicine, College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University. Female, 8-week-old BALB/c mice were purchased from Experimental Animal Center of Harbin Veterinary Research Institute (Chinese Academy of Agricultural Sciences, Harbin, China).…”

“…Previously the nucleotide sequence of the predicted immunodominant region of bovine haptoglobin (pirBoHp), removing the signal peptide sequence, was synthesized based on the codon usage bias of Escherichia coli. (9) The synthesized pirBoHp gene was successfully expressed in E. coli BL21 (DE3) cells. The polyclonal antibody against the recombinant pirBoHp protein could recognize aand b-chains of the native bovine haptoglobin.…”