【摘要】 目的利用噬菌体表面呈现技术研制高活性、特异性抗骨肉瘤干扰素。方法将干扰素IFNαlc／86DDNA插入噬菌体质粒载体pCANTAB5E，并构建噬菌体IFNαlc／86DAB环突变文库；通过在OS732细胞上竞争性亲合洗脱、MTT法对比检测抗肿瘤活性，筛选针对OS732细胞的高活性噬菌体-IFN突变体。结果获得2个噬菌体-IFN突变体，其抗OS732细胞增殖活性较突变母体分别提高4和16倍。结论αl型干扰素可在噬菌体表面正确折叠表达。这一技术可用于研制高活性特异性IFN。更多还原

【Abstract】 Objective To develop some specific anti-osteosarcoma (OS) interferons (IFNs) with high activity. Methods First, theIFN-ale/86D DNA was fused to a DNA sequence of phagemid vector-pCANTABSE. A phage-IF’Nalc/86D library was thengenerated by fully randomizing the sequence of the four positions 29,31,32,and 35 in AB-loop. Secondly, a new and feasible phagedisplay panning, and competitive affinity selection was developed. Some variants with high sanity to OS cells were obtained. MTTcolorimetric assay was used to explore the activity of these variants and phage-IFNalc/86D in regulating cell groWth of OS732.Results Two new phage-IFNalc/86D variants were selected, whose anti-proliferation on OS cell line was 4- and 16-fold higher thantheir parental wild type phage-IFN. Conclusions The IF’Nalc/86D can be correctly folded on the surface of phage. This techniquecan be used to develop specific IFNs with high biological activity.更多还原