REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY ASSAY FOR RECOMBINANT ACIDIC FIBROBLAST GROWTH-FACTOR IN ESCHERICHIA-COLI CELL-SUSPENSIONS AND LYSATE SAMPLES

被引：8

作者：

DEPHILLIPS, P

BUCKLAND, B

GBEWONYO, K

YAMAZAKI, S

SITRIN, R

机构：

[1] Merck Research Labs, West Point, PA 19406

来源：

JOURNAL OF CHROMATOGRAPHY A | 1994年 / 663卷 / 01期

关键词：

D O I：

10.1016/0021-9673(94)80494-X

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A reversed-phase HPLC assay for acidic Fibroblast Growth Factor (aFGF) expressed in E. coli is described. The assay was developed on a Polymer Labs. PLRP-S macroporous poly(styrene-benzene) column. A sample preparation procedure was developed to permit quantitation of aFGF in crude samples, such as cell suspensions and cell lysates. The assay was linear over the range of 25-140 mu g/ml with an accuracy of 5%. By using a POROS column containing a stationary phase with ''through-pores'' and a superficial mobile phase velocity of 1440 cm/h, the analysis could be performed in 3 min. The polymeric supports used for this assay were durable; periodic washing with sodium hydroxide maintained column performance for extended periods.

引用

收藏

页码：43 / 51

页数：9

相关论文

共 13 条

[1] PERFUSION CHROMATOGRAPHY PACKING MATERIALS FOR PROTEINS AND PEPTIDES [J].

AFEYAN, NB ;

FULTON, SP ;

REGNIER, FE .

JOURNAL OF CHROMATOGRAPHY, 1991, 544 (1-2) :267-279

[2] FLOW-THROUGH PARTICLES FOR THE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC SEPARATION OF BIOMOLECULES - PERFUSION CHROMATOGRAPHY [J].

AFEYAN, NB ;

GORDON, NF ;

MAZSAROFF, I ;

VARADY, L ;

FULTON, SP ;

YANG, YB ;

REGNIER, FE .

JOURNAL OF CHROMATOGRAPHY, 1990, 519 (01) :1-29

[3] SEPARATION OF PROTEINS BY REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY .1. OPTIMIZING THE COLUMN [J].

BURTON, WG ;

NUGENT, KD ;

SLATTERY, TK ;

SUMMERS, BR ;

SNYDER, LR .

JOURNAL OF CHROMATOGRAPHY, 1988, 443 :363-379

[4] DETECTION OF ALPHA-1-ANTITRYPSIN FROM RECOMBINANT ESCHERICHIA-COLI LYSATES UTILIZING THE PARTICLE CONCENTRATION FLUORESCENCE IMMUNOASSAY [J].

DELTITO, BJ ;

ZABRISKIE, DW ;

ARCURI, EJ .

JOURNAL OF IMMUNOLOGICAL METHODS, 1988, 107 (01) :67-72

[5]

DONG M W, 1989, Biochromatography, V4, P19

[6] VERY HIGH-SPEED SEPARATION OF PROTEINS WITH A 20-MU-M REVERSED-PHASE SORBENT [J].

FULTON, SP ;

AFEYAN, NB ;

GORDON, NF ;

REGNIER, FE .

JOURNAL OF CHROMATOGRAPHY, 1991, 547 (1-2) :452-456

[7] A HETEROGENEOUS DOUBLE ANTIBODY ENZYME-LINKED IMMUNOASSAY TO MEASURE BETA-GALACTOSIDASE FUSION PROTEIN [J].

HAYASHIBE, K ;

SASSANO, D ;

ZIAI, MR .

JOURNAL OF IMMUNOASSAY, 1990, 11 (01) :89-95

[8] PROTEIN RELEASE FROM ESCHERICHIA-COLI-CELLS PERMEABILIZED WITH GUANIDINE-HCI AND TRITON X100 [J].

HETTWER, D ;

WANG, H .

BIOTECHNOLOGY AND BIOENGINEERING, 1989, 33 (07) :886-895

[9] SEPARATION OF PROTEINS BY REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY .2. OPTIMIZING SAMPLE PRETREATMENT AND MOBILE PHASE CONDITIONS [J].

NUGENT, KD ;

BURTON, WG ;

SLATTERY, TK ;

JOHNSON, BF ;

SNYDER, LR .

JOURNAL OF CHROMATOGRAPHY, 1988, 443 :381-397

[10] A REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC METHOD FOR CHARACTERIZATION OF BIOSYNTHETIC HUMAN GROWTH-HORMONE [J].

RIGGIN, RM ;

DORULLA, GK ;

MINER, DJ .

ANALYTICAL BIOCHEMISTRY, 1987, 167 (01) :199-209