Immunoglobulin light chain classes in a teleost fish.
Immunoglobulin light chain classes in a teleost fish.
复制标题
硬骨鱼中的免疫球蛋白轻链类别。
作者：
C. J. Lobb;M. Olson;L. Clem
通过SDS-PAGE对癍点叉尾鮰的主要的、约700，000道尔顿的血清抗体（Ab）进行分析，揭示了重（H）链的仅一种分子量种类（约70，000道尔顿），而轻（L）链的相对迁移率的显著异质性是明显的。观察到约26，000、约24，000和约22，000道尔顿的三种分子量L链变体。直到开发出与鲶鱼免疫球蛋白（IG）反应的小鼠单克隆抗体（mAb）后，L链变异体之间的关系才清楚。其中两个单克隆抗体，命名为3F12和1G7，被发现独立地识别不同群体的鲶鱼IG。此外，当这两种mAb组合使用时，它们免疫沉淀大于95%的特异性纯化的鲶鱼抗DNP或抗荧光素Ab。发现从与mAb 3F12或1G7缀合的免疫吸附亲和基质中回收的鲶鱼IG的L链明显不同。单克隆抗体3F12与含有约24，000和约22，000道尔顿L链变体的鲶鱼IG亚群反应，而mAb 1G7与仅含有约26，000道尔顿L链变体的鲶鱼IG另一亚群反应。固相板结合试验与轻度减少鲶鱼IG H和L链的使用表明，这两种mAb优先与鲶鱼L链反应。随后的分析表明，这两个抗原性不同的L链群体的肽图谱程序表明，这些L链的结构不同。此外，从个别鲶鱼的血清分析表明，两个L链类型的存在下，在每25鲶鱼检查。这些研究强烈表明，不同的L链类已经在鱼类中进化。
Analysis of predominant, approximately 700,000 dalton serum antibody (Ab) of the channel catfish by SDS-PAGE revealed only one molecular mass species of heavy (H) chain (approximately 70,000 daltons), whereas a marked heterogeneity in the relative mobilities of the light (L) chains was evident. Three molecular mass L chain variants of approximately 26,000, approximately 24,000, and approximately 22,000 daltons were observed. The relationships between the L chain variants were not clear until mouse monoclonal Ab (mAb) reactive with catfish immunoglobulin (Ig) were developed. Two of these mAb, designated 3F12 and 1G7, were found to independently recognize different populations of catfish Ig. In addition, when these two mAb were used in combination they immunoprecipitated greater than 95% of specifically purified catfish anti-DNP or anti-fluorescein Ab. The L chains of catfish Ig recovered from immunoabsorbent affinity matrices conjugated with either mAb 3F12 or 1G7 were found to be clearly distinct. Monoclonal Ab 3F12 reacted with a subpopulation of catfish Ig that contained the approximately 24,000 and approximately 22,000 dalton L chain variants, whereas mAb 1G7 reacted with another subpopulation of catfish Ig that contained only the approximately 26,000 dalton L chain variant. Solid phase plate binding assays with the use of mildly reduced catfish Ig H and L chains showed that both of these mAb preferentially reacted with catfish L chains. Subsequent analysis of the two antigenically distinct L chain populations by peptide mapping procedures demonstrated that these L chains were structurally different. Furthermore, analysis of the serum from individual catfish showed the presence of both L chain types in each of 25 catfish examined. These studies strongly suggest that distinct L chain classes have evolved in fish.