硫氰酸盐洗脱法测定噬菌体抗体的相对亲和力

Relative affinity measurement of phage antibodies by ELISA using thiocyanate elution

目的 建立一种测定噬菌体抗体相对亲和力的方法。方法 参照硫氰酸盐洗脱法测定完整抗体分子和Fab段相对亲和力的方法 ,在ELISA实验中以酶标抗M13为二抗检测 5个单克隆噬菌体抗体的相对亲和力 ,并与可溶性Fab段的相对亲和力进行比较。结果 噬菌体抗体可以耐受硫氰酸盐的洗脱 ,用硫氰酸盐洗脱法测定 5个噬菌体抗体所得亲和力排序与测定其相应可溶性抗体分子片段所得结果一致。结论 硫氰酸盐洗脱法可用于噬菌体抗体相对亲和力的测定。

Objective To establish a method for the measurement of relative affinity of phage antibodies. Methods The relative affinities of 5 phage antibodies were measured by ELISA using thiocyanate elution. A HRP-conjugated anti-M13 was used as secondary antibody, and the affinity of the phage antibodies were compared with that of the Fab fragments. Results Phage antibodies were tolerant to thiocyanate elution. The relative affinity of 5 phage antibodies measured by ELISA using thiocyanate elution coincided with that of their corresponding Fab fragments. Conclusion The relative affinity of phage antibodies could be determined by ELISA using thiocyanate.