Kasheverov I E, Golovnina T A, Iarov A V, Utkin Iu N, Tsetlin V I
Bioorg Khim. 1994 Jul;20(7):720-30.
The peptides which correspond to the fragments 55-64, 182-192, 225-236 and 236-245 (P1-P4, respectively) of the rat brain substance P receptor were synthesized. Antibodies (Ab1-Ab4, respectively) against the KLH-conjugates of these peptides were raised and purified by affinity chromatography. None of the antibodies inhibited the Bolton-Hunter labelled substance P, [125I]BH-SP, binding to the rat brain membranes. On the other hand, Ab2 and Ab3 recognition of the SP receptor was found in ELISA experiments: the CHAPS--solubilized rat brain membranes could inhibit binding of these antibodies to the immobilized P2 and P3. Antibodies Ab1-Ab4 did not interact with the CHAPS-solubilized DDS cross-linked complex of the [125I]BH-SP and SP receptor. However, this complex retained the capacity of interacting with the affinity-purified antibodies against SP and was purified by sequential gel-permeation HPLC and protein A-chromatography.
合成了与大鼠脑P物质受体的55 - 64、182 - 192、225 - 236和236 - 245片段(分别为P1 - P4)相对应的肽段。针对这些肽段的KLH缀合物制备了抗体(分别为Ab1 - Ab4),并通过亲和层析进行纯化。这些抗体均未抑制博尔顿 - 亨特标记的P物质([¹²⁵I]BH - SP)与大鼠脑膜的结合。另一方面,在ELISA实验中发现Ab2和Ab3可识别SP受体:经CHAPS增溶的大鼠脑膜能够抑制这些抗体与固定化的P2和P3的结合。抗体Ab1 - Ab4不与经CHAPS增溶的[¹²⁵I]BH - SP和SP受体的DDS交联复合物相互作用。然而,该复合物保留了与抗SP的亲和纯化抗体相互作用的能力,并通过连续凝胶渗透HPLC和蛋白A层析进行了纯化。