المستخلص: |
Histidine-rich glycoprotein HRG which is an abundant serum protein of the cystatin superfamily. It combines a varied array of ligands containing blood clotting systems and components of the immune and is important for many physiological and pathological processes in cell adhesion, cell migration, fibrinolysis, coagulation, complement activation, and angiogenesis. In addition, HRG has been enhancing the clearance of necrotic cells. The histidine-rich region HRR is showing as the domain of HRG in regulating tumour angiogenesis and promoting tumour progression.The antibody to HRR human is important for the study role of HRG in cancer and therapy. The primary aim of this project was produce recombinant human HRR which is in bacteria by using the pHUE expression system and purify with Ni-NTA chromatography. Also, another aim of project is generated monoclonal antibody against His-HRR . Expression of HRR was performed in Bl21 (DE3) with different concentration IPTG and temperature. It was done with 1 mM IPTG and 0.1 mM IPTG at 20 oC and 37 oC. The purification was done with His tag by FPLC. The sample was prepared to inject into three mice. The first bleed was tested by ELISA and western blot. One of mouse was possible contain monoclonal antibody against His-HRR. Every bleed will be testing in coming time.
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