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Index 341
Genenase I, 170, 214, 215, 217, 225
Glutathione, 125–127, 131, 133
concentration, 133
ligand attachment, 132
-S transferase, 38
-Streamline matrix, production of, 128
GST
activity assay, 131
fusion protein, 126, 134
HaloTag, 191–202, 204, 207
advantages of, 194
binding characteristics of, 195
protocol for immobilization of, 195, 197
High-throughput purification, 163–164
High-yield protein expression system, 153, 162
Hill plot equation is, 107
His-tag protein
purification of, 44, 140–141, 235
sequence, alternative, 231
HisLink binding, 157, 164
spin column, 152, 156–157, 165
Histidine tag, 25–26, 39, 48, 137–140, 229–230,
232, 234, 238, 241
Homo-bifunctional ligands, 39
Homo sapiens, 295
HQ tag proteins, 151–152, 155–156, 158–160,
162–165
purification of, 151, 154, 160–163
Human immunoglobulins, 53
Human rhinovirus, 215
Hydrogen donor, 242
Hydrophilic
chromatography, 1
interactions, 6, 46, 132, 173, 224–225
resins, 74
IMA chromatography, 41
Iminodiacetic acid, 26, 40, 138
Immobilization
efficiency, 257–258, 261, 269
method, 259–260, 262, 269, 273
of molecules, 247
of process enzymes, 230
of proteins, 193
Immobilized affinity metal chromatography, 151
Immobilized enzyme, 262
Immobilized gluthathione ligands, 276
Immobilized Metal Affinity Chromatography,
25–27, 33–34, 38, 40, 48, 75, 137, 138–140,
146–147, 151–152, 230–231, 233, 235–238,
240, 242, 248, 252–253, 285–286
Immobilized metal chelate complex (IMCC),
26–27, 33–34, 138, 140, 142, 146–147
Immobilized phosphatidylcholine column, 297
Immobilized Phospholipid Chromatography, 295
Immunoaffinity chromatography, 53, 55–56
chromatogram for, 57
Insect and mammalian cells, 160
Ion exchange chromatography, 72
Ionic detergent, 217, 219–220, 225
Isoforms, resolution of, 13
Isolation
buffer, 309
of a peptide, 113
process, 296
Kunitz hyperchromicity assay, 265
Laser-induced fluorescence detection
principles, 313
Ligand density measurement, 128
Ligand utilization, 132, 258
Lipid-based transfection reagent, 204
Liquid chromatography, 8, 25, 309
high performance, 72, 222, 304
Low-affinity inhibitors, 4
Lower critical solution temperature (LCST), 40
Lymphocytes, 249–250, 252–253
Lysis Buffer, 155–156
Lysis of Pelleted Bacterial Cells, 155, 157
MagneHis protein purification system,
152–153, 164
Magnetic nickel purification, 152–153
Maltodextrin-binding protein, 170
Maltose-binding protein (MBP), 13, 169
Maltose regulon, 171, 173–174, 184
Mammalian cell culture, 53–54, 61, 68, 160, 213
Mapping of ligand-binding sites, 314
Mass spectrometry, 13, 164, 314, 316
elution conditions, 154
Matrix-assisted dialysis refolding methods, 178, 182
Maxwell purification instrument, 163
Membrane proteins, 140, 171, 295–296, 301
Metal affinity precipitation technique, 49
Metal chelate affinity chromatography, 38, 286
Metal copolymer, recycling of the, 45
Michaelis–menten constant, 262, 266, 273
Micropipettor, 288
Mobility shift ACE, 305, 318–320, 325
Molecular biology, 234
Molecular interactions, 304–305
Monoclonal antibody, 53, 54, 111, 113, 117, 320