Purification of human embryonic and semi-embryonic hemoglobins.

(A) Human Hb Gower-1 (ζ₂ε₂). (Left) Hemolysate prepared from adult mα^+/−/mβ^−/−/hζ/hε complex transgenic–knockout mice was resolved over a Poros SP/H column using a linear 20 to 120 mM NaCl gradient (pH 6.5). The positions of human Hb ζ₂ε₂ (peak 1) and contaminant hybrid mouse–human Hb mα₂hε₂ (peak 2) are indicated (arbitrary A₂₈₀ units). Eluate conductivity (in mS) is depicted by a gray line. (Right) Aliquots of unfractionated (U) lysate and eluate corresponding to peaks 1 and 2 were resolved by nondenaturing cellulose acetate electrophoresis. A control lane contains a mixture of human Hbs A, F, S, and C. The migration of constituent hemoglobins is indicated to the left, and gel polarity to the right. (B) Human Hb Gower-2 (α₂ε₂). Resolution of hemolysate from an adult mα^+/−/mβ^−/−/hα/hε complex transgenic–knockout mouse into human Hb α₂ε₂ (peak 2) and contaminant hybrid mouse/human Hb mα₂hε₂ (peak 1) using a nonlinear 60- to 100-mM NaCl gradient (pH 6.8). (C) Human Hb Portland-2 (ζ₂β₂). Resolution of hemolysate from an adult mα^+/−/mβ^−/−/hζ/hβ complex transgenic–knockout mouse into human Hb ζ₂β₂ (peak 1) and contaminant hybrid mouse/human Hb mα₂hβ₂ (peak 2) using a linear 20- to 80-mM NaCl gradient (pH 6.5).