原子力显微镜对常用7种单克隆抗体形态的观察

Morphologies of seven commonly used monoclonal antibodies under atomic force microscope

  • 摘要: 目的 为抗原抗体反应提供原子力显微镜下的形态学基础,为建立常用抗体形态结构的形态库打下基础。 方法 将7种常用单克隆抗体分别滴加于载玻片上,放入加有10%中性缓冲甲醛的湿盒内,置于37 ℃温箱中1 h,用蒸馏水轻轻涮洗,晾干,用原子力显微镜(atomic force microscope, AFM)进行扫描,观察形态,测量扫描颗粒的宽度、最大峰高度和平均粗糙度。 结果 在2D图中,细胞角蛋白(广谱)、S100蛋白鼠抗人单克隆抗体为长杆状,波形蛋白、白细胞共同抗原、细胞角蛋白7、Dog-1鼠抗人单克隆抗体为类圆形,甲状腺球蛋白鼠抗人单克隆抗体为三角形;不同组间单克隆抗体之间宽度、最大峰高度的差异有统计学意义(P< 0.05),各组间平均粗糙度的差异无统计学意义(P> 0.05)。 结论 2D图中,7种单克隆抗体的形态大致可分为三种:长杆状、类圆形、三角形,直观地印证了抗体具有相对特异性,并为抗原的共同表达和例外表达的进一步研究提供形态学基础。

     

    Abstract: Objective To lay the foundation for establishing a morphological structure database of commonly used antibodies by studying the morphology of antigen-antibody reaction under atomic forced microscope. Methods Seven antibodies were added onto slides and placed into a wet kit containing 10% neutral buffer formalin. One hour after the antibodies were kept in an incubator at 37 ℃, they were washed with distilled water and dried. Their morphologies were observed under atomic force microscope. The width, maximum height and mean roughness of particles were measured. Results The long rod cytokeratin and S100 protein muse anti-human monoclonal antibodies, the oblate round common antigens of vimentin and leukocyte as well as cytokeratin 7 and Dog-1 mouse antihuman monoclonal antibodies, and the triangle thyroglobulin monoclonal antibodies were observed in 2D figures. The width and maximum height of monoclonal antibodies were significantly different (P< 0.05), but no significant difference was found in the mean roughness (P> 0.05). Conclusion The morphologies of 7 monoclonal antibodies can be divided into long rod, oblate round and triangle types, which directly prove the relative specificity of antibodies and can thus provide the morphological foundation for further studying the co-expression and exceptional expression of antigens.

     

/

返回文章
返回