Background: AAV2 vector containing sFLT01, with the ability of VEGF and PLGF neutralization, was delivered into oxygen induced retinopathy (OIR) mouse model in 2008. The recombinant fusion protein represented appreciated amount of anti angiogenic effects. FLT1 (VEGFR1) is a cell surface receptor for VEGF and PLGF. Alternative splicing in flt1 gene produces soluble form (s FLT1) which circulates in blood flow and neutralizes excess VEGF and PLGF. In 2 other studies, it was recognized that the second extracellular domain of flt1 plays the crucial role in binding to VEGF and PLGF which includes amino acids no. 124-229. sFLT01 contains signal peptide of flt1 gene, domain 2 of sflt1, a short 9 glycin linker and gamma immunoglobulin Fc region . Purpose: According to the pharmaceutical and commercial worth of this medicine, we targeted it’s fabrication as a biosimilar process in Iran. So at first step we used bioinformatics competence to predict and design the sequence of gene of interest.

By referring to nucleotide database of NCBI, The interested variant was selected as the soluble form. The signal peptide sequence is coding by 286-363th nucleotides. We considered the 286th nucleotide as the first nucleotide of the gene and in respect to this nucleotide; we approached to 124th amino acid. It corresponds to the 654-972th nucleotides. This would be the nucleotides composition of the second domain. To determine the equivalent protein sequence of the selected nucleotide sequences, we used EMBOSS Transeq of EBI database. It predicted the equivalent amino acid sequence of the selected nucleotides.

We used NCBI retrieved sequences to design our home made sFLT01 sequence. We evaluated the designed sequence for the amino acids coded and determined that the selected sequence would be efficiently translated to interested neutralizing protein in experimental procedure.

This sequence was submitted for synthesis and would be manipulated to be cloned in AAV-2 gene delivery system in the next experimental procedures.