Purification and Analysis of Folate-Dye Conjugate

Folate receptor has achieved great importance for use in targeted drug delivery and in vitro and in vivo diagnostic assays. Folate receptor (FR) is over expressed on several cancer cell types and activated macrophages, but exists in relatively low abundance on the surface of non-cancerous tissue and non-inflammatory cells. This unique attribute is the hallmark of folate – targeted methodologies allowing the diagnostic agent and/or drug to be ferried to the specific disease sites while preventing “off target” effects in normal tissues. Although several folate-based dyes have been reported, simple and detailed synthetic and purification methods for these conjugates have not been reported. Our objective is to develop a new folate-based dye and the corresponding HPLC methods for the purification and analysis of this conjugate. Folate- DylightTM 800 was successfully synthesized by very simple, inexpensive and safe method. The whole synthesis method can be performed in small laboratories with minimum resources and small quantity of reagents required. The synthesis of Folate-DylightTM 800 conjugate was successfully purified by HPLC. The purified Folate-DylightTM 800 conjugate was evaluated by microplate reader, UV-Vis and HPLC.