Figure 2.

Physiochemical characteristics of HM-3-Fc. (1) Purity analysis of HM-3-Fc (A). Columns: TOSOH, TSKgel G3000SWxL PN: 008543, Mobile Phase: 0.4 M NaCl 0.03 M Na₂HPO₄ 0.02 M NaH₂PO₄ Solution with 10% Acetonitrile. Isocratic elution, Flow: 0.5 mL/min, Temperature: 25 °C, Detection fluorescence (FL): 214 nm, Injected volume: 10 μL. (2) CE-SDS analysis of HM-3-Fc. Non-reduced capillary electrophoresis (CE) analysis (B) and reduced CE-SDS analysis (C). Sample concentration: 1 mg/mL heated in water bath at 70 °C for 10 min. The sample was injected to the anode with reverse polarity using −5 kV for 20 s. Separation voltage was −15 kV with reverse polarity during the 30 min separation. Detection wavelength was 214 nm. (3) cIEF analysis of HM-3-Fc. Holograph of cIEF profile (D) and amplified part of cIEF graph (E). B1 = basic peak 1, A1–A4 = acidic peaks 1–5. The samples were detected by Beckman MDQ capillary electrophoresis system using a neutral coated capillary. Inject sample for 99.0 s at 20 psi. Focusing: 15 min at 25 kV under normal polarity. Mobilization: 30 min at 30 kV under normal polarity. Detection wavelength was 280 nm.