Williams K M, Williams J, Marshall T
Analytical Biochemistry Research Group, School of Health Sciences, The University of Sunderland, Great Britain.
Electrophoresis. 1998 Jul;19(10):1828-35. doi: 10.1002/elps.1150191047.
Analysis of Bence Jones proteinuria by high resolution two-dimensional electrophoresis (2-DE) and immunoblotting reveals a complex pattern of light chain (LC) isoforms corresponding to the free monoclonal Bence Jones protein and its fragments. Replica blotting gives duplicate blots for LC typing (lambda, chi) and, under the conditions employed, leaves sufficient protein for Coomassie Blue staining of the urinary protein profile and pIIMr determination of the LC isoforms. Carrier ampholytes (CAs, in our "simplified" 2-DE system) and immobilised pH gradients (IPGs, in the Multiphor 2-DE system) give similar LC isoform patterns. Artifacts, including cone-like distortions and trailing "piggyback" spots, are visualised with both 2-DE systems. IPGs are advantageous as they allow reproducible detection of strongly basic LC isoforms by isoelectric focusing (under equilibrium conditions) without recourse to CA nonequilibrium pH gradient electrophoresis.
通过高分辨率二维电泳（2-DE）和免疫印迹法分析本斯·琼斯蛋白尿，揭示了与游离单克隆本斯·琼斯蛋白及其片段相对应的轻链（LC）亚型的复杂模式。复制品印迹法可得到用于轻链分型（λ链、κ链）的重复印迹，并且在所采用的条件下，剩余足够的蛋白质用于考马斯亮蓝染色尿蛋白图谱以及轻链亚型的pIIMr测定。载体两性电解质（在我们的“简化”二维电泳系统中）和固定化pH梯度（在Multiphor二维电泳系统中）给出相似的轻链亚型模式。两种二维电泳系统都能观察到包括锥形畸变和拖尾“搭便车”斑点在内的假象。固定化pH梯度具有优势，因为它们允许通过等电聚焦（在平衡条件下）可重复检测强碱性轻链亚型，而无需采用载体两性电解质非平衡pH梯度电泳。
