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首页> 外文会议>ICoMST 2011;International conference of meat science and technology >Characterization of Antibodies against Connectin/Titin 20-kDa Fragment Increased in Chicken Sarcoplasm during Postmortem Aging
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Characterization of Antibodies against Connectin/Titin 20-kDa Fragment Increased in Chicken Sarcoplasm during Postmortem Aging

机译：死后衰老过程中鸡肌浆中针对Connectin / Titin 20 kDa片段增加的抗体的表征

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In this study, polvclonal and monoclonal antibodies produced against connectin/titin 20-kDa fragment, which increased in chicken sarcoplasm with postmortem aging, were characterized in order to utilize for estimating meat aging. Recombinant 12- and 16-kDa fragments of internal sequence of the 20-kDa fragment were prepared as immunogens. The result of ELISA with rat anti-12-kDa fragment antiserum indicated that the absorbance was higher with sarcoplasm of aged chicken muscle than that of immediately postmortem muscle, and an indirect immunofluorescence microscopy showed that the antiserum bound to the Z-line of sarcomeres. Two stable hybridoma cell lines (16-4B4 and 1-3C) were produced with mouse spleen cell immunized by recombinant 12-kDa fragment and myeloma cell (P3U1). Isotypes of monoclonal antibodies (fhAbs) 16-4B4 and 1-3C secreted from the fused hybridoma cells were IgG3 and IgG2b with κ-light chain, respectively and both mAbs were specific for the 20-kDa fragment increased in chicken sarcoplasm with postmortem aging time. While, the results of western blotting proved that the antibodies cross-react to the bands in sarcoplasm from aged pork and beef muscles similarly with chicken sarcoplasm. Therfore, it is concluded that the antibodies against connectin/titin 20-kDa fragment are capable of use for estimating meat aging.
机译：在这项研究中，针对连接蛋白/ titin 20-kDa片段产生的polvclonal和单克隆抗体的特性进行了表征，这些片段在鸡尸体中随着死后老化而增加，以用于估计肉类老化。制备20-kDa片段内部序列的重组12-和16-kDa片段作为免疫原。用大鼠抗12-kDa片段抗血清进行ELISA的结果表明，老年鸡肌肉的肌浆中的吸光度比立即死后的肌肉要高，间接免疫荧光显微镜检查显示抗血清与肉瘤的Z线结合。用重组12-kDa片段免疫的小鼠脾细胞和骨髓瘤细胞（P3U1）产生了两种稳定的杂交瘤细胞系（16-4B4和1-3C）。融合杂交瘤细胞分泌的单克隆抗体（fhAbs）16-4B4和1-3C的同种型分别是带有κ轻链的IgG3和IgG2b，并且两种单克隆抗体都对鸡肌浆中20kDa片段具有特异性，并随死后老化而增加。 。同时，蛋白质印迹法的结果证明，抗体与老猪肉和牛肉肌肉的肌浆中的条带发生交叉反应，与鸡肌浆相似。因此，得出的结论是，针对连接蛋白/视黄素20-kDa片段的抗体能够用于估计肉的衰老。

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