Bin Song K, Won M, Meares C F
Department of Food Science and Technology, Chungnam National University, Taejon 305-764, Korea.
Biotechnol Appl Biochem. 1998 Oct;28 ( Pt 2):163-7.
Lym-1 single-chain Fv (sFv) can be used for targeted radiodiagnosis and therapy of B-lymphocytic malignancies. Lym-1 sFv was constructed and expressed as a glutathione S-transferase fusion protein, using a (G4S)3 linker connecting the C-terminus of the VH domain and the N-terminus of the VL domain of Lym-1. Six histidine residues and an E Tag epitope were introduced at the C-terminus of the sFv. Lym-1 sFv was purified with glutathione-Sepharose 4B affinity chromatography followed by digestion with thrombin. Lym-1 sFv of 28 kDa was confirmed by Western blotting with anti-(E Tag) monoclonal antibody. An antigen binding assay of Lym-1 and a CD study indicated that it is functionally active.
Lym-1单链Fv（sFv）可用于B淋巴细胞恶性肿瘤的靶向放射诊断和治疗。构建了Lym-1 sFv并将其表达为谷胱甘肽S-转移酶融合蛋白，使用(G4S)3接头连接Lym-1的VH结构域的C末端和VL结构域的N末端。在sFv的C末端引入了六个组氨酸残基和一个E标签表位。通过谷胱甘肽-琼脂糖4B亲和色谱法纯化Lym-1 sFv，然后用凝血酶消化。用抗（E标签）单克隆抗体进行蛋白质免疫印迹证实了28 kDa的Lym-1 sFv。Lym-1的抗原结合试验和圆二色性研究表明它具有功能活性。
