Preparation for the Monoclonal Antibody to the High Molecular Weight Proteins in the Myofibrils

肌原纤维高分子蛋白单克隆抗体的制备

• 批准号: 62560281
• 负责人: SUZUKI Atsushi
• 金额: $ 1.34万
• 依托单位: Niigata University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62560281/
• 关键词: Muscle Protein; Myofibril; Meat Tenderization; Connectin; Z-nin; Monoclonal Antibody; ポリクロナル抗体; 筋腹線維; Z-nin; コネクチンのモノクロナル抗体

Connection and Z-nin are high molecular weight proteins in the myofibrils. There are some questions about the role of both proteins on meat tenderization during postmortem storage.1. The isolation of native connectih from rabbit skeletal muscle (L.dorsi) was carried out, and postmortem changes in native connection were studied quantitatively and qualitatively. On the basis of the amino acid composition and electrophoretic mobility on sds-polyacrylamide gel, it is ahown that the isolated protein was highly pure connectin. An increase of the extractability of native connectin from the muscle during storage was observed, and an electron microscopic study of the purified native connection indicates a qualitative change of connectin in the muscle during postmortem storage. If connection has some influence on meat tenderness, the quantitative and qualitative change of native connectin in the muscle are possibly one of the cause of meat tenderzation during cunditioning.2. The polyclonal antibody to the native connectin was obtained from the balb/c mouse immunized with chicken connectin. The spleen cells of the balb/c mouse were fused with mouse myeloma cells (line NS-1) and the screening of the hybridoma cell with an ability to make antibody to connectin is now underway. From these results the procedure to obtain the monoclonal antibody to the native connectin seems to be almost established.3. As to the Z-NIN, the isolation of it in a native state is impossible at present. So the ployclonal antibody to the denatured Z-NIN was prepared in our laboratory.

Connection和Z-nin是肌原纤维中的高分子量蛋白。关于这两种蛋白质在肉死后贮藏过程中嫩化的作用还存在一些问题。从家兔骨骼肌（L.dorsi）中分离出天然连接体，并对其死后的变化进行了定量和定性研究。根据氨基酸组成和sds-聚丙烯酰胺凝胶的电泳迁移率，表明分离的蛋白为高纯度的连接蛋白。在储存过程中，观察到天然连接蛋白从肌肉中可提取性的增加，并且对纯化的天然连接蛋白的电镜研究表明，在死后储存过程中，肌肉中的连接蛋白发生了质的变化。如果连接对肉的嫩度有一定的影响，那么肌肉中天然连接蛋白的量变和质变可能是调理过程中肉嫩化的原因之一。从鸡连接蛋白免疫的balb/c小鼠中获得了天然连接蛋白的多克隆抗体。将balb/c小鼠的脾脏细胞与小鼠骨髓瘤细胞（NS-1系）融合，目前正在筛选具有产生连接蛋白抗体能力的杂交瘤细胞。从这些结果来看，获得天然连接蛋白单克隆抗体的方法似乎基本建立起来了。对于Z-NIN，目前在原生状态下对其进行隔离是不可能的。因此，我们在实验室制备了变性Z-NIN的多克隆抗体。

项目成果

Suzuki, Atsushi: "Postmortem Changes of Native Connectin in Rabbit Skeletal Muscle" Agricultural and Biological Chemistry. 52. 1439-1444 (1988)

Suzuki Atsushi：“兔骨骼肌中天然连接素的死后变化”农业和生物化学。

Suzuki, Atsushi: Recent Studies on Meat Conditioning: in Advances in Food Science and Technology (III). Korin, 77-96 (1988)

Suzuki Atsushi：肉类调理的最新研究：食品科学与技术的进展（III）。

鈴木敦士: "畜肉の熟成に関する研究の進歩-解硬・軟化機構を中心に-食品工業における科学・技術の進歩(III)" 光琳, 77-96 (1988)

Atsushi Suzuki：“畜肉老化的研究进展 - 关注裂解和软化机制 - 食品工业的科学技术进展（III）”Korin，77-96（1988）

Atsushi,Suzuki: Agriculturel and Biological Chemistry. 52. 1439-1444 (1988)

Atsushi，Suzuki：农业和生物化学。

Atsushi SUZUKI: Agricultural and Biological Chemistry. 52. (1988)

铃木敦：农业和生物化学。