sFLT01 is an engineered chimeric protein with the inhibitory effect on VEGF and PLGF which are expressed aberrantly in AMD ocular disease. There was no exact information about its nucleic acid or protein sequence in litreture. So at first we approached to determine it’s sequence. By referring to an article that had been published in 1997 we found crystal structure of VEGF in a complex with domain 2 of FLT1 receptor.Then we achieved to the FLT1 second domain amino acid sequence and thereafter corresponding nucleotide sequence was determined.Using EMBOSS Transeq tool of EBI database and NCBI BLAST tool our proposed sequence got validated. Finally Modeling servers and softwares applied for the proposed sequence structure validation

In this study we determined tertiary structure of our proposed sequence for this protein in its chimeric form and compared it with constitutive components in natural forms. To predict tertiary structure,we used Modeling servers such as I-TASSER, Swiss Model,Phyre2 and PsiPred. The prediction results analyzed with PDBviewer and PyMOL softwares. We transfected cultured hRPE cells by the expression construct and demonstrated anti angiogenic function of cells' conditioned media by in vitro angiogenesis method.

Software analysis demonstrated that,our proposed chimeric construct which was derived from natural proteins sequences (sFLT1 and IgG1) was in an excellent accordance with its natural counterpart based on its tertiary structure and preserved its natural folding exactly. In Vitro angiogenesis assay results showed decreased potential of tube formation in conditioned medium of treated RPE cultures, by constructs expressing Sflt01, when compared with control cultures

In conclusion our desired sFLT01 sequence could produce functional neutralizing protein receptor for VEGF/PlGF