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目的 探讨在不同的免疫学检测方法中 ,与单链抗体相连的亲和标记肽对单链抗体与抗原结合活性的影响。方法 在制备“VH 连接链 VL”型结构抗乙酰胆碱受体单链抗体时 ,将c myc标记肽与单链抗体的VL 连接 ,应用先将抗c myc抗体固定单链抗体的固相放射免疫测定法、直接将单链抗体与乙酰胆碱受体结合的液相放射免疫测定法 ,分别测定单链抗体与乙酰胆碱受体的结合活性。结果 在固相放射免疫测定法中 ,单链抗体不能与乙酰胆碱受体结合 ;在液相放射免疫测定法中 ,单链抗体则能与乙酰胆碱受体结合。结论 在应用不同的免疫学检测方法中 ,与单链抗体相连的c myc标记肽可影响单链抗体与抗原的结合活性。
Objective To investigate the effect of single-chain antibody-conjugated affinity tagged peptides on the binding activity of single chain antibodies to antigen in different immunological detection methods. Methods The c-myc-tagged peptide was linked to the VL of single-chain antibody in the preparation of “VH Link VL” structure anti-acetylcholine receptor single-chain antibody. The solid-phase radioimmunoassay Method, direct single-chain antibody and acetylcholine receptor liquid phase radioimmunoassay, single chain antibody and acetylcholine receptor binding activity were measured. Results In solid-phase radioimmunoassay, single-chain antibodies can not bind to acetylcholine receptors; in liquid radioimmunoassay, single-chain antibodies bind to acetylcholine receptors. Conclusion In different immunological methods, the c-myc-labeled peptide linked to single-chain antibody can affect the binding activity of single-chain antibody to antigen.