【摘要】 为制备猪骨骼肌肌球蛋白轻链激酶(skMLCK)的特异性单克隆抗体,设计合成多肽并与载体蛋白偶联,经细胞融合、培养基筛选、阳性细胞的检测与单克隆化后,获得3株能稳定分泌抗skMLCK单克隆抗体的杂交瘤细胞株;种植BALB/c小鼠腹腔,收集腹水,纯化后得到skMLCK的单克隆抗体。分别应用高效液相色谱、质谱和Western blot对纯化单克隆抗体进行抗体纯度、效价和敏感性检测。结果显示:获得的猪skMLCK单克隆抗体,能特异性识别skMLCK纯化蛋白、猪肌肉组织的内源性蛋白。本试验成功制备了能特异性结合猪skMLCK的单克隆抗体,这为今后进一步研究探讨skMLCK对猪肌肉发育的影响提供物质基础。更多还原

【Abstract】 To prepare and identify the specific monoclonal antibody of porcine skeletal muscle myosin light chain kinase(skMLCK),peptides were synthesized and conjugated to carrier proteins.Monoclonal antibodies are made by cell culture that involves fusing myeloma cells with mouse spleen cells immunized with the desired antigen.High performance liquid chromatography,mass spectrometry and Western blot were used to detect the purity,titer and sensitivity of purified monoclonal antibody.The results showed that the monoclonal antibody of porcine skMLCK was obtained.Purified antibody can specifically recognize purified skMLCK and endogenous protein of porcine muscle tissue.This study provides a material foundation for further research on the effects of skMLCK on muscle development in pigs.更多还原