Isolation of human growth hormone isohormones D and E in milligram amounts (I), using isotachophoresis on polyacrylamide gel.
Human growth hormone (hGH) isohormones D and E were isolated from a plasmin digest of 150 mg hGH. Isotachophoresis on a cylindrical polyacrylamide gel of 18 mm diameter served to separate the various isohormone species. The "extended stack" was sliced, and slices containing hGH-D and -E were extracted by Steady-State Stacking on 1% agarose gel and collected in a 1 ml volume. The extract was purified on gel chromatography and lyophilized. The overall yield was approximately 19 mg of hGH (D + E) and 16 mg of hGH-(D, minor E). This represents a 62% recovery of the amount of hGH-D and -E in the digest estimated densitometrically. The purity of the product, based on Lowry analysis, specific RIA or UV-absorbance, was 91%. The isolated hGH-(D+E) exhibited an enhanced bioactivity in the rat tibia line assay.
D000818:Animals; D001681:Biological Assay; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005341:Fibrinolysin; D013006:Growth Hormone / Q000302:isolation & purification*; D006801:Humans; D008832:Microchemistry / Q000379:methods; D010446:Peptide Fragments / Q000302:isolation & purification*; D011863:Radioimmunoassay; D051381:Rats; D013056:Spectrophotometry, Ultraviolet
D010446:Peptide Fragments; C030484:human growth hormone isohormone D; C030485:human growth hormone isohormone E; D013006:Growth Hormone; D005341:Fibrinolysin