Suphioglu C, Singh M B, Knox R B
School of Botany, University of Melbourne, Parkville, Australia.
Int Arch Allergy Immunol. 1993;102(2):144-51. doi: 10.1159/000236565.
Group I allergens, from eight different clinically important grass pollens of the Pooideae (rye-grass, canary, Kentucky bluegrass, orchard and timothy), Chloridoideae (Bermuda grass) and Panicoideae (Johnson grass, maize) were isolated by sodium dodecyl sulphate polyacrylamide gel electrophoresis and subjected to limited partial proteolysis and cyanogen bromide cleavage. The generated digests were visualised on gels by silver staining. Replicate gels were blotted and screened with Lolp I-specific mAb FMC A1 and IgE binding to an allergic human serum pool, to determine the degree of antigenic and IgE-binding similarities, respectively. The highest antigenic and IgE-binding similarities were between orchard and rye-grass pollens, both in the same tribe Poeae, and among the closely related grass genera. No peptide mapping of major grass pollen allergens has previously been undertaken. This study proves peptide mapping is a powerful method for the preliminary identification and internal amino acid microsequencing of common antigenic and IgE-binding determinants of allergens, providing information that is useful in the development of reagents for the treatment of grass-pollen-associated allergies.
第一组变应原，从早熟禾亚科（黑麦草、金丝雀草、草地早熟禾、果园草和梯牧草）、虎尾草亚科（狗牙根）和黍亚科（约翰逊草、玉米）的8种临床上重要的不同草花粉中，通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳分离出来，并进行有限的部分蛋白酶解和溴化氰裂解。产生的消化产物通过银染在凝胶上可视化。复制凝胶进行印迹，并用黑麦草I特异性单克隆抗体FMC A1和与过敏性人血清池结合的IgE进行筛选，分别确定抗原性和IgE结合相似性的程度。果园草和黑麦草花粉之间的抗原性和IgE结合相似性最高，它们都属于同一黑麦草族，且在亲缘关系密切的草属中。此前尚未对主要草花粉变应原进行肽图谱分析。本研究证明，肽图谱分析是一种强大的方法，可用于变应原常见抗原性和IgE结合决定簇的初步鉴定和内部氨基酸微测序，为开发治疗草花粉相关过敏的试剂提供有用信息。
