人骨形成蛋白2A与小鼠成骨细胞表面特异性结合的研究

Specific Binding of Human BMP2A with a Mouse Osteoblastic Cell MC3T3

作者证实在大肠杆菌中表达的人骨形成蛋白２Ａ（ｈＢＭＰ２Ａ）Ｃ端肽段具有骨诱导活性，再利用噬菌体呈现载体克隆并表达了ｈＢＭＰ２ＡｃＤＮＡ３’端５６７个核苷酸，表面呈现有人ＢＭＰ２ＡＣ端肽段的噬菌体与小鼠成骨细胞系（ＭＣ３Ｔ３）能够特异地结合，用抗Ｍ１３噬菌体抗体进行ＥＬＩＳＡ实验呈阳性反应，同时用３ＨＴｄＲ参入重组ＢＭＰ２Ａ噬菌体时放射性计数比对照组高近１０倍，推测小鼠成骨细胞表面存在有人ＢＭＰ２Ａ受体。

Human BMP2A C terminal peptide was expressed in E. coli system and the expressed product was able to induce ectopic bone formation in mouse muscle. Then its cDNA 3'terminal 567 nucleotides were cloned into a phage display vector(pCSM21) and expressed as phage g3p fusion protein on the phage surface. thymidine was incorporated into the recombinant phage DNA. The mouse osteoblastic cells (MC3T3) were coated onto the surface of well in a 96 well plastic microtiter plate. The recombinant phage BMP2A was added to the wells coated by MC3T3, and allowed to interact with the cells. The MC3T3 cells bound with recombinant phage BMP2A showed a positive result in ELISA. The TdR counts of the sample groups were 3 to 10 times higher than the control groups. It suggests that the surface of MC3T3 cells present the receptor for human BMP2A.