Table 1.
List of buffers, composition and recipes used for chromatography steps.
| Buffer | Buffer composition | Buffer stock solutionsa |
|---|---|---|
| Buffer-A | 24 mM HEPES, pH 6.8 | Line A1 – 100 mM HEPES |
| Line A2 – 100 mM NaOH | ||
| Line B1 – ddH2O | ||
| Line B2 – 2 M NaCl | ||
| Buffer-B | 24 mM HEPES; 1 M NaCl, pH 6.8 | Line A1 – 100 mM HEPES |
| Line A2 – 100 mM NaOH | ||
| Line B1 – ddH2O | ||
| Line B2 – 2 M NaCl | ||
| Buffer-C | 7.8 mM Na2HPO4; 150 mM NaCl, pH 7.5; | Line A1 – 30 mM Na2HPO4 |
| Line A2 – 100 mM HCl | ||
| Line B1 – ddH2O | ||
| Line B2 – 2 M NaCl | ||
| Buffer-D | 15.5 mM 1-methyl-piperazine; 15.5 mM Bis–Tris; 7.8 mM Tris, pH 8.5 | Line A1 – 50 mM 1-methyl-piperazine; 50 mM Bis–Tris; 25 mM Tris |
| Line A2 – 100 mM HCl | ||
| Line B1 – ddH2O | ||
| Line B2 – 2 M NaCl | ||
| Buffer-E | 15.5 mM 1-methyl-piperazine; 15.5 mM Bis–Tris; 7.8 mM Tris; 1 M NaCl, pH 8.5 | Line A1 – 50 mM 1-methyl-piperazine; 50 mM Bis–Tris; 25 mM Tris |
| Line A2 – 100 mM HCl | ||
| Line B1 – ddH2O | ||
| Line B2 – 2 M NaCl | ||
| Buffer-F | 5.1 mM Na2HPO4; 5.1 mM Formate Na; 10.2 mM Acetate Na, pH 5.5 | Line A1 – 30 mM Na2HPO4; 30 mM Formate Na; 60 mM Acetate Na |
| Line A2 – 100 mM HCl | ||
| Line B1 – ddH2O | ||
| Line B2 – 2 M NaCl | ||
| Buffer-G | 5.1 mM Na2HPO4; 5.1 mM Formate Na; 10.2 mM Acetate Na; 1 M NaCl, pH 5.5 | Line A1 – 30 mM Na2HPO4; 30 mM Formate Na; 60 mM Acetate Na |
| Line A2 – 100 mM HCl | ||
| Line B1 – ddH2O | ||
| Line B2 – 2 M NaCl | ||
| Buffer-H | 25 mM Tris; 25 mM NaCl; 0.5 mM EDTA, 10% glycerol, pH 7.5 | Manually prepared |
a
Buffer recipe stock solutions and the ratio mixtures for the final buffer composition were generated as described in the UNICORN (v5.11, GE Healthcare) operating software.