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人自然杀伤细胞表面的抑制受体（ＫＩＲ），是与表达在其他细胞表面的相应ＨＬＡＩ类分子相互作用的。许多ＫＩＲ受体已被克隆，小部分量的单克隆抗体也已产生。然而为区分不同位点及其等位基因的产物，并精确阐明其功能，需要更多的研究试剂。本研究探讨用噬菌体抗体库，分离ＫＩＲ单链抗体（ｓｃＦｖ）的可行性。方法：ＮＫＡＴ２－ＩｇＧ１为一ＫＩＲ，与人ＩｇＧ１Ｆｃ以融合蛋白形成表达而获得。噬菌体抗体库为“Ｎｉｓｉｍ”抗体库。抗体库ＮＫＡＴ２－ＩｇＧ１包被的免疫管５次筛选。ｓｃＦｖ由特异的噬菌体感染细菌，诱导而得。筛选效应经噬体菌体的菌落形成单位及ＤＮＡ酶切图谱检测。筛选的克隆进行ＤＮＡ序列分析。ｓｃＦｖ的特异性及免疫学特征由ＥＬＩＳＡ、聚丙烯胺凝胶电泳及免疫印迹分析。细胞结合实验由流式细胞仪测试。结果：５次筛选后，ＮＫＡＴ２特异的噬菌体达到２００倍的富集，ＤＮＡ酶切图谱也显示某种特定消化类型的富集。经ＥＬＩＳＡ分析，结果显示３７个克隆与ＮＫＡＴ２反应呈强阳性。这些克隆与ＮＫＡＴ１－ＩｇＧ１、ＮＫＡＴ３－ＩｇＧ１、ＮＫＡＴ４－ＩｇＧ１、人ＩｇＧ及其他非相关蛋白的反应均为阴性。其中一个克隆（ｓｃＨＵＢ１）经流式细胞仪进一步测试，显示?
Human killer cell surface inhibitory receptors (KIRs) interact with the corresponding HLAI-like molecules expressed on the surface of other cells. Many KIR receptors have been cloned and a small amount of monoclonal antibody has also been produced. However, in order to differentiate the products of different loci and their alleles and to elucidate their function precisely, more research reagents are needed. This study explored the feasibility of using phage antibody libraries to isolate KIR single chain antibodies (scFv). Methods: NKAT2-IgG1 was a KIR and was expressed as a fusion protein with human IgG1Fc. The phage antibody library is a “Nisim” antibody library. Antibody library NKAT2-IgG1 coated immune tube 5 times screening. The scFv is infected by specific bacteriophages and induced. The screening effect was detected by colony forming units of phage phage and DNA digestion map. Selected clones were subjected to DNA sequence analysis. The specificity and immunological characteristics of scFv were analyzed by ELISA, polyacrylamide gel electrophoresis and immunoblotting. Cell binding assays were tested by flow cytometry. Results: After 5 rounds of screening, the NKAT2-specific phage reached a 200-fold enrichment and the DNA digestion pattern also showed enrichment for a particular digestive type. By ELISA analysis, the results showed that 37 clones and NKAT2 reaction was strongly positive. The responses of these clones to NKAT1-IgG1, NKAT3-IgG1, NKAT4-IgG1, human IgG and other non-related proteins were all negative. One of the clones (scHUB1) was further tested by flow cytometry to show that