Cottam G P, Wheeler A W, Cook R M, Moran D M
Bencard Allergy Unit, Beecham Pharmaceutics Biosciences Research Centre, Epsom, Surrey, England, U.K.
Immunol Lett. 1988 Apr;17(4):345-9. doi: 10.1016/0165-2478(88)90009-0.
Two fragment pools, one of MW greater than 10,000 and the other of MW between 1,000 and 10,000, were prepared by the sequential treatment of rye grass pollen extract with cyanogen bromide (cleavage at Met-X) and 2-nitro-5-thiocyanobenzoic acid (cleavage at X-Cys). Electrophoretic analysis of the two pools showed that none of the major components of the whole extract remained intact. Characterisation of the two fragment pools by radioimmunoassay showed that whilst they both lost the ability to bind to human anti-rye IgE antibodies, they largely retained their reactivity towards both human and mouse anti-rye IgG antibodies. In addition, the higher molecular weight pool retained its ability to stimulate extract-specific T cells, after accessory cell processing. This separation of the immunological properties of rye grass pollen extract by chemical cleavage is seen as a basis for the development of novel immunotherapy agents.
通过用溴化氰（在甲硫氨酸-X处裂解）和2-硝基-5-硫氰基苯甲酸（在X-半胱氨酸处裂解）对黑麦草花粉提取物进行顺序处理，制备了两个片段库，一个分子量大于10000，另一个分子量在1000至10000之间。对这两个库的电泳分析表明，整个提取物的主要成分均未保持完整。通过放射免疫分析对这两个片段库进行表征表明，虽然它们都失去了与人抗黑麦IgE抗体结合的能力，但它们在很大程度上保留了对人和小鼠抗黑麦IgG抗体的反应性。此外，经过辅助细胞处理后，较高分子量的库保留了刺激提取物特异性T细胞的能力。通过化学裂解分离黑麦草花粉提取物的免疫特性被视为开发新型免疫治疗剂的基础。
