Nguyen N Y, Grindeland R E, Chrambach A
Prep Biochem. 1981;11(2):173-89. doi: 10.1080/00327488108064240.
Human growth hormone (hGH) isohormones D and E were prepared from a partial enzymatic hydrolyzate catalyzed by plasmin, using isoelectric focusing on polyacrylamide gel to fractionate the digest. Separation between the two isohormones species was optimized by employing a maximally flattened pH gradient between pH's 4.3 and 5.6. Gel slices containing isohormones D and E were extracted and concentrated by Steady-State Stacking on polyacrylamide gel. The extract was purified by gel filtration and lyophilized. Overall yields of lyophilized isohormones D and E were 10.9 mg and represented a recovery of 51% relative to the densitometrically estimated isohormone concentrations in the hGH digest. Purity of the products, based on Lowry analysis, U.V. absorbance and radioimmunoassay (RIA) was 60-70%. The isolated isohormone species were active in the rat tibia-line assay and in the specific RIA for hGH.
人生长激素（hGH）同工激素D和E是通过纤溶酶催化的部分酶水解产物制备的，采用聚丙烯酰胺凝胶等电聚焦对消化产物进行分级分离。通过在pH值4.3至5.6之间采用最大程度平坦的pH梯度，优化了两种同工激素之间的分离。含有同工激素D和E的凝胶切片通过聚丙烯酰胺凝胶上的稳态堆积进行提取和浓缩。提取物通过凝胶过滤纯化并冻干。冻干的同工激素D和E的总产率为10.9毫克，相对于hGH消化物中通过光密度法估计的同工激素浓度，回收率为51%。基于洛瑞分析、紫外吸光度和放射免疫测定（RIA），产物的纯度为60-70%。分离出的同工激素在大鼠胫骨线试验和hGH的特异性RIA中具有活性。
