Zhao Hong-Liang, Xue Chong, Xiong Xiang-Hua, Zhang Wei, Yang Bing-Fen, Liu Zhi-Min
Beijing Institute of Biotechnology, Beijing 100071, China.
Sheng Wu Gong Cheng Xue Bao. 2005 Mar;21(2):254-8.
To increase the in vivo half-life of human CNTF mutein AX15 (R13K), HSA-AX15 (R13K) fusion protein was constructed by the fusion of the C-terminus of HSA to the N-terminus of AX15 (R13K) via an 11 amino acids linker. HSA-AX15 (R13K) fusion protein was purified to homogeneity by cation exchange chromatography, reverse phase chromatography and gel filtration after expressed in pichia pastoris. TF-1 cell survival bioassay showed the biological activity of AX15 (R13K) was not affected by the fusion to HSA. It was demonstrated that tertian injection of 4.8 mg/kg HSA-AX15 (R13K) fusion protein could produce more potent anti-obesity effects on KM mice than daily injection of 1.6 mg/kg AX15 (R13K). The long-acting form of hCNTF variant has the potential to reduce discomfort by requiring fewer injections and to minimize the side-effects by decreasing the dosage and fluctuation of plasma concentration, and thus has superior clinical application.
为了延长人睫状神经营养因子突变体AX15（R13K）的体内半衰期，通过一个11个氨基酸的接头将人血清白蛋白（HSA）的C末端与AX15（R13K）的N末端融合，构建了HSA-AX15（R13K）融合蛋白。在毕赤酵母中表达后，通过阳离子交换色谱、反相色谱和凝胶过滤将HSA-AX15（R13K）融合蛋白纯化至均一。TF-1细胞存活生物测定表明，AX15（R13K）与HSA融合后其生物活性不受影响。结果表明，每三天注射4.8mg/kg的HSA-AX15（R13K）融合蛋白对KM小鼠产生的抗肥胖作用比每天注射1.6mg/kg的AX15（R13K）更强。hCNTF变体的长效形式有可能通过减少注射次数来减轻不适，并通过降低血浆浓度的剂量和波动来最小化副作用，因此具有卓越的临床应用价值。
