Carding S R, Thorpe R, Childs R A, Spitz M, Feizi T
Biochem J. 1984 May 15;220(1):253-60. doi: 10.1042/bj2200253.
With the aim of obtaining monospecific antibodies against the beta-galactoside-binding lectin of bovine heart muscle, spleen cells from Lou rats immunized with lectin were fused with the rat myeloma line Y3.Ag1.2.3. Two immunoglobulin M (IgM)-producing clones, designated NIBy 142-36/8 and NIBy 143-9/5, derived from separate fusions, were used to generate ascites containing high-titre binding activity against the 13kDa component in preparations of lectin. Direct evidence that haemagglutinating activity is associated with the 13kDa protein was obtained by the specific elution of 13kDa polypeptides with haemagglutinating activity from an immobilized antibody adsorbent. Solid-phase radiobinding assays and immunoblotting of isolated lectins and/or muscle homogenates confirmed the earlier indications with conventional antisera that the beta-galactoside-binding lectins of bovine, human and monkey muscle tissue are antigenically related.
为了获得针对牛心肌β-半乳糖苷结合凝集素的单特异性抗体，用凝集素免疫的Lou大鼠的脾细胞与大鼠骨髓瘤细胞系Y3.Ag1.2.3进行融合。从单独融合中获得的两个产生免疫球蛋白M（IgM）的克隆，分别命名为NIBy 142-36/8和NIBy 143-9/5，用于产生腹水，该腹水对凝集素制剂中的13kDa成分具有高滴度结合活性。通过从固定化抗体吸附剂中特异性洗脱具有血凝活性的13kDa多肽，获得了血凝活性与13kDa蛋白相关的直接证据。固相放射结合测定以及对分离的凝集素和/或肌肉匀浆的免疫印迹证实了早期用传统抗血清得出的结论，即牛、人及猴肌肉组织中的β-半乳糖苷结合凝集素在抗原性上相关。
