N-[(carbamoylmethyl)amino]ethanesulfonic acid improves phenotyping of alpha 1-antitrypsin by isoelectric focusing on agarose gel.

The hereditary deficiency variants of alpha 1-antitrypsin that are associated with diseases such as emphysema are usually identified by use of isoelectric focusing on polyacrylamide gels. Agarose is a simpler, faster, safer, and more reliable medium for this, but resolution often is not as good. I describe a method in which the ultrathin agarose gel contains N-[(carbamoylmethyl)amino]ethanesulfonic acid as a "separator," to flatten the pH gradient and improve separation of the alpha 1-antitrypsin isoforms. The resolution obtained equals or surpasses that of conventional methods based on use of either polyacrylamide or agarose. Haptoglobin, which interferes with isoelectric focusing on polyacrylamide, does not interfere with this method; other advantages are also discussed.