猫CD44基因的表达及其单克隆抗体的制备与鉴定OA北大核心
Expression of feline CD44 gene and generation and identification of its monoclonal antibodies
为制备用于实验的猫CD44单克隆抗体,原核表达了猫CD44重组蛋白,通过镍亲和层析法纯化目的蛋白,并将蛋白定量至1 mg/mL,然后将蛋白免疫BALB/c小鼠,通过杂交瘤抗体技术制备鼠抗猫CD44单抗,并验证单抗的适用范围.结果 表明,成功制备并纯化4株猫CD44单抗,其效价均高于1∶64000,重链均为IgG1型,轻链均为κ型.制备的单抗可应用于Western-blot、间接免疫荧光和免疫组织化学等实验,为猫CD44基因相关疾病的研究和药物研发等提供了重要的单克隆抗体资源.
To generate experimental monoclonal antibodies against feline CD44,recombinant feline CD44 protein was expressed in prokaryotic systems,and was purified using nickel affinity chromatography to a concentration of 1 mg/mL,and was immunized BALB/c mice.The murine anti-feline CD44 monoclonal anti-body was generated by the hybridoma antibody technique,and the scope of application of the monoclonal anti-body was verified.Four monoclonal antibodies were successfully generated and purified.These antibo-dies demonstrated high titers(＞1∶64 000),with uniform IgG1 heavy chains and κ light chains.These mono-clonal antibodies can be applicable in Western-blot,indirect immunofluorescence,and immunohisto-chemistry assays,thereby providing valuable tools for studying CD44-related diseases and drug deve-lopment in feline models.
乔亚楠;王秀羽;惠晓晨;颜权辉;李秋燕
华中农业大学动物医学院农业微生物资源发掘与利用全国重点实验室,湖北武汉 430070华中农业大学动物医学院农业微生物资源发掘与利用全国重点实验室,湖北武汉 430070华中农业大学动物医学院农业微生物资源发掘与利用全国重点实验室,湖北武汉 430070华中农业大学动物医学院农业微生物资源发掘与利用全国重点实验室,湖北武汉 430070华中农业大学动物医学院农业微生物资源发掘与利用全国重点实验室,湖北武汉 430070
畜牧业
猫CD44原核表达单克隆抗体制备鉴定
felineCD44prokaryotic expressionmonoclonal antibodygenerationidentification
《中国兽医科学》 2025 (5)
616-624,9
国家重点研发计划项目(2023YFD1800302)湖北省试验动物研究项目(2023CFA005)
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