Affinity purification of alpha-galactosidase A from human spleen, placenta, and plasma with elimination of pyrogen contamination. Properties of the purified splenic enzyme compared to other forms.

摘要: The substrate analog alpha-D-galactosylamine was synthesized, linked to 6-aminohexanoic acid, and coupled carboxyhexyl-Sepharose. This affinity support permitted the purification of human alpha-galactosidase A (alpha-D-galactoside galactohydrolase, EC 3.2.1.22) from spleen, placenta, plasma. When used in conjunction with conventional procedures, chromatography enabled rapid specific each source. Significantly, pyrogenic endotoxins were eliminated enzyme preparations by use column. Splenic purified high yield (38%) a activity 1.9 X 10(6) units/mg. homodimer native molecular weight 101,000 subunit 49,800. UV absorption coefficient E280 1% = 18 lambda max 282 nm. plasma form markedly improved (229,000 units/mg) which 3 times greater than that achieved previously. enzymes plasma, placenta immunologically identical. physical kinetic properties consistent confirmed previous findings.