SEPARATION OF CARBOHYDRATE-MEDIATED MICROHETEROGENEITY OF RECOMBINANT-HUMAN-ERYTHROPOIETIN BY FREE SOLUTION CAPILLARY ELECTROPHORESIS - EFFECTS OF PH, BUFFER TYPE AND ORGANIC ADDITIVES

被引：109

作者：

TRAN, AD ^{[1
]}

PARK, SG ^{[1
]}

LISI, PJ ^{[1
]}

HUYNH, OT ^{[1
]}

RYALL, RR ^{[1
]}

LANE, PA ^{[1
]}

机构：

[1] ORTHO PHARMACEUT CORP,RW JOHNSON PHARMACEUT RES INST,DEPT ANALYT RES & DEV,RARITAN,NJ 08869

来源：

JOURNAL OF CHROMATOGRAPHY | 1991年 / 542卷 / 02期

关键词：

D O I：

10.1016/S0021-9673(01)88781-1

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Free solution capillary electrophoresis has been investigated as an alternative to isoelectric focusing for the separation of the glycoforms of recombinant human erythropoietin (r-HuEPO), a primary regulator of erythropoiesis. A systematic approach was used to study the effect of pH, buffer type and organic modifiers on the resolution of the microheterogeneity of erythropoietin. The main factors for improving the resolution were the regulation of the electroosmotic flow of the running buffer and the reduction of solute-wall interaction. The best resolution of the glycoforms of r-HuEPO was obtained with a mixed buffer pH 4.0 (100 mM acetate-phosphate, 10 h preequilibration time).

引用

页码：459 / 471

页数：13

共 15 条

[1]

COHEN AS, 1988, J CHROMATOGR, V448, P41

[2] A SEMIEMPIRICAL MODEL FOR THE ELECTROPHORETIC MOBILITIES OF PEPTIDES IN FREE-SOLUTION CAPILLARY ELECTROPHORESIS [J].

GROSSMAN, PD ;

COLBURN, JC ;

LAUER, HH .

ANALYTICAL BIOCHEMISTRY, 1989, 179 (01) :28-33

[3]

HIRTH P, 1988, CONTRIB NEPHROL, V66, P50

[4]

HJERTEN S, 1989, J CHROMATOGR, V492, P585

[5] ISOLATION AND CHARACTERIZATION OF GENOMIC AND CDNA CLONES OF HUMAN ERYTHROPOIETIN [J].

JACOBS, K ;

SHOEMAKER, C ;

RUDERSDORF, R ;

NEILL, SD ;

KAUFMAN, RJ ;

MUFSON, A ;

SEEHRA, J ;

JONES, SS ;

HEWICK, R ;

FRITSCH, EF ;

KAWAKITA, M ;

SHIMIZU, T ;

MIYAKE, T .

NATURE, 1985, 313 (6005) :806-810

[6] CAPILLARY ZONE ELECTROPHORESIS [J].

JORGENSON, JW ;

LUKACS, KD .

SCIENCE, 1983, 222 (4621) :266-272

[7] FAST AND HIGH-RESOLUTION ANALYSIS OF HUMAN-SERUM TRANSFERRIN BY HIGH-PERFORMANCE ISOELECTRIC-FOCUSING IN CAPILLARIES [J].

KILAR, F ;

HJERTEN, S .

ELECTROPHORESIS, 1989, 10 (01) :23-29

[8]

LAI PH, 1986, J BIOL CHEM, V261, P3116

[9] CAPILLARY ZONE ELECTROPHORETIC SEPARATION OF PEPTIDES AND PROTEINS USING LOW PH BUFFERS IN MODIFIED SILICA CAPILLARIES [J].

MCCORMICK, RM .

ANALYTICAL CHEMISTRY, 1988, 60 (21) :2322-2328

[10]

Righetti P. G., 1983, ISOELECTRIC FOCUSING

← 1 2 →