Preparation and identification of monoclonal antibodies against bovine haptoglobin

来源 :中国畜牧兽医学会家畜内科学会2014年学术研讨会 | 被引量 : 0次 | 上传用户:yusiyuangame
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  Haptoglobins are members of the alpha 2-globulin family of proteins, mainly expressing in the liver, skin,lung, kidney and adipose tissue.Haptoglobin functions to bind free plasma hemoglobin, preventing oxidative damage.Additionally, haptoglobin is an acute-phase protein ,whose expression level increases in the inflammatory process.The accumulating reports indicated that bovine haptoglobin (BoHp) was a potential biomarker in many inflammatory diseases of dairy cows caused by infectious microorganisms, involving in footrot, mastitis, enteritis, peritonitis,endocarditis, abscesses,endometritis,interdigital dermatitis, and so on.Thus, BoHp has the potential use as an early diagnostic marker of inflammatory diseases in dairy cattle.In order to provide some basis for development of rapid diagnostic reagents of BoHp,female 8-week-old BALB/c mice were immunized with purified recombinant proteins of the predicted immunodominant region of bovine haptoglobin (pirBoHp).A total of two monoclonal antibodies (McAbs) ,named 1B3 and 6D6, were prepared by conventional B lymphocyte hybridoma technique.Tites of ascites fluid and cell-cuhure supernatant of the McAb 1B3 were 1∶9.6 × 108 and 1∶8.2 × 104 ,respectively ,and that of the the McAb 6D6 were 1∶4.4 × 105 and 1∶1.0 × 104, respectively.The subtype of the McAbs 1B3 and 6D6 was IgG1κ.In western blot,the McAbs 1B3 and 6D6 could recognize α-chain of the native BoHp from plasma of dairy cows.These data indicated that the McAbs 1B3 and 6D6 will have a potential use for developing diagnostic reagents of Bo Hp.
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