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- W2379098206 abstract "Objective To express and purify vascular endothelial growth factor 165(VEGF165)protein and prepare its monoclonal antibody(McAb).Methods The recombinant plasmid pGEX-4T-1 / VEGF165 constructed previously was transformed to E.coli BL21(DE3)pLysS for expression of GST-VEGF165 fusion protein under induction of IPTG.The expressed product was dis-solved with sarkosyl and purified by affinity chromatography.McAb was prepared by immunizing BALB / c mice with the purified GST-VEGF165 fusion protein,identified by Western blot and analyzed for subtype.An double antibody sandwich ELISA kit was de-veloped with the prepared McAb and evaluated for linearity range.Results The expressed GST-VEGF165 fusion protein,with a rel-ative molecular mass of 45 000,contained about 20% of total somatic protein,reached a purity of more than 90% after purification and showed an affinity of about 109 L / mol to its receptor.A hybridoma cell strain secreting McAb against GST-VEGF165 was ob-tained.The secreted McAb,with good reactogenicity as proved by Western blot,was identified as IgG2a and showed affinity to VEGF165(Kdis 2×10-9 mol / L).The linearity range of ELISA kit prepared with the secreted McAb was 1.562 5 ～ 25 ng / ml.Conclusion VEGF165 protein was successfully expressed and purified,and its McAb was prepared,which laid a foundation of development of detection kit for VEGF." @default.
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- W2379098206 date "2008-01-01" @default.
- W2379098206 modified "2023-09-22" @default.
- W2379098206 title "Expression of Vascular Endothelial Growth Factor Gene and Preparation of Monoclonal Antibody against Expressed Product" @default.
- W2379098206 hasPublicationYear "2008" @default.
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