Effect of detergent on electromigration of proteins: CE of very low density lipoprotein receptor modules and viral proteins.
The different electrophoretic behavior of the members of two groups of proteins with respect to the absence or presence of detergent additives in the BGE was explored. Recombinant soluble concatemers of repeat 3 of the very low density lipoprotein (VLDL)-receptor fused at their N-terminus to maltose-binding protein (MBP) exhibited different electrophoretic mobilities in borate buffer (pH 8.3) in the absence and in the presence of dodecyl-PEG ether (D-PEG). This enabled the separation of the receptor fragments from MBP after enzymatic cleavage. In the presence of SDS, the mobilities of all proteins approached the same values with increase in detergent concentrations. In contrast, viral capsid proteins of a human rhinovirus (HRV) exhibited different migration in the presence of the additive. For the receptor proteins, extreme apparent high plate numbers were observed when the SDS concentration in the sample and the separation buffer differed. This effect might be erroneously interpreted as a high efficiency. However, it is due to the conductivity boundaries caused by the sample and leads to a total loss of separation.
Kremser
Leopold
L
Institute for Analytical Chemistry, University of Vienna, Vienna, Austria. Leopold.kremser@univie.ac.at
Bilek
Gerhard
G
Kenndler
Ernst
E
D001665:Binding Sites; D002021:Buffers; D002213:Capsid / Q000737:chemistry*; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification; D003902:Detergents / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D006801:Humans; D058619:Maltose-Binding Proteins; D011485:Protein Binding; D011973:Receptors, LDL / Q000302:isolation & purification*; D012229:Rhinovirus / Q000737:chemistry; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry*; D014764:Viral Proteins / Q000737:chemistry / Q000302:isolation & purification*
D002021:Buffers; D002352:Carrier Proteins; D003902:Detergents; D058619:Maltose-Binding Proteins; D011973:Receptors, LDL; C036053:VLDL receptor; D014764:Viral Proteins; D012967:Sodium Dodecyl Sulfate