Effect Of Human Fibroblast Growth Factor 2 Nanobody On Stability Of FGF-2

Protein and peptide drugs have the advantages of high specificity and low toxicity.In recent years,protein and peptide drugs have become research hotspots in the field of medicine,but they have problems such as poor stability and critical storage condition.It is important to solve the problem of the stability of protein drugs for application.As an important cytokine in human body,FGF-2 has the potential to develop into many types of protein drugs,but its protein is highly prone to coagulation reaction and inactivation.The aim of this study is using Nb4 combine with FGF-2 to enhance the stability without affecting its biological activity.High-density cell fermentation and NiRTA affinity chromatography purification technology are used to obtain Nb4 and Nb6 proteins with a purity of more than 95%.The FGF-2 protein was obtained by shake flask fermentation and CM-Sephadex Fast Flow ion exchange chromatography and Heparin-sepharose CL-6B affinity chromatography.0.2 mg/mL of a mixture of FGF-2 and Nb4(FGF-2+Nb4)and a control group of 0.2 mg/mL FGF-2 were obtained.The two types of samples are treated at different temperatures(4°C,25°C,37°C,40°C);pH(5,7,9,11);placement time(1 to 4 weeks);additives(protein protectants and denaturation)agent;rotating speed(500rpm,1000rpm).Western Blotting and ELISA are used for testing.Cell proliferation is used for verification.The result is that FGF-2 is extremely unstable under extreme conditions such as high temperature and alkali;Nb4 is more protective against FGF-2 in extreme environments;The best effect of FGF-2+Nb4 sample when 1 mol/L sucrose is used as auxiliary additive which can be combined as a protein protectant candidate.