膜联蛋白 A5（AnnexinA5）特异性结合磷脂酰丝氨酸，具有重要的生理功能。作为脊椎动物膜联蛋白家族一员，最早是从人的胎盘分离得到的。膜联蛋白 A5 的高级结构已经解析清楚，但对于其生物学功能特点仍然有待深入研究。本论文基于前期实验室研究基础，基于亲和层析技术和特异性蛋白质酶切技术，首先对该重组蛋白质进行了分离纯化。进而使用 SDS-PAGE 凝胶电泳技术对纯化产物进行了检测与鉴定，使用 BCA 方法测定纯化蛋白浓度。进而，本论文使用该重组蛋白质对小鼠进行多次免疫，成功制备出特异性抗膜联蛋白的多克隆抗体。ELISA 实验检测抗体效价良好（给出数值）。基于我们制备出的抗膜联蛋白多克隆抗体，对膜联蛋白及衍生物的磷脂分子结合特征进行了检测与研究。研究表明，膜联蛋白及衍生物具备良好的磷脂酰丝氨酸结合特异性。基于细胞粘附实验，研究了膜联蛋白及衍生物的整合素受体结合特征。进而，我们还通过构建 CAM 模型，探讨了膜联蛋白及衍生物的血管增生生物学效应，并进行了初步的数据分析，研究表明，膜联蛋白及衍生物表现出了对于新生血管的抑制效应。相关研究为后续进一步深入探讨膜联蛋白及衍生物的作用机制提供了初步的实验数据参考。

Annexin A5 specifically binds to phosphatidylserine and has important physiological functions. As a member of the vertebrate membrane-bound protein family, it was initially isolated from human placenta. The higher order structure of Annexin A5 has been resolved, but its biological functional characteristics still need further study. Based on the previous laboratory research and using affinity chromatography and specific protein cleavage technology, this paper first purified the recombinant protein. Subsequently, purified products were detected and identified by using SDS-PAGE gel electrophoresis technology, and the protein
concentration was measured using the BCA method. Furthermore, this paper used the recombinant protein to immunize mice multiple times and successfully prepared polyclonal antibodies specific to Annexin A5. ELISA experiments detected good antibody potency (numerical value provided). Based on the polyclonal antibody we prepared against Annexin A5, the phospholipid binding characteristics of Annexin A5 and its derivatives were studied and detected. The study showed that Annexin A5 and its derivatives had good phosphatidylserine binding specificity. Based on cell adhesion experiments, the integrin receptor binding characteristics of Annexin A5 and its derivatives were studied.
Furthermore, we explored the neovascularization inhibition biological effect of 4 Annexin A5 and its derivatives by constructing a CAM model and conducted preliminary data analysis. The study showed that Annexin A5 and its derivatives showed inhibitory effects on neovascularization. This research provides a preliminary experimental data reference for further exploring the mechanisms of action ofAnnexinA5 and its derivatives.