Sodium dodecyl sulfate-polyacrylamide gel electrophoresis for screening nonaggregating human antibody heavy chain variable domains.
Antibody heavy chain variable domains (V(H)s) form a significant class of biologics. With V(H) display libraries-the primary source of V(H) binders-unwanted aggregating V(H)s are coselected, sometimes overwhelmingly, alongside nonaggregating V(H)s. Thus, methods enabling efficient screening for nonaggregating V(H)s are highly valuable. Here, we found that on nonreducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels, nonaggregating V(H)s migrate faster than expected, giving underestimated molecular weights (MWs), whereas aggregating ones migrate slower, giving overestimated MWs. Our finding can be applied to large-scale screening for nonaggregating V(H)s and possibly other proteins, in particular in display library settings, by SDS-PAGE.
Kim
Dae Young
DY
Institute for Biological Sciences, National Research Council Canada, Ottawa, Ontario, Canada K1A 0R6.
Tanha
Jamshid
J
D004591:Electrophoresis, Polyacrylamide Gel / Q000191:economics / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000191:economics / Q000379:methods; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry*; D007135:Immunoglobulin Variable Region / Q000737:chemistry*
D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region