PURIFICATION AND DETERMINATION OF GLUTAMINE-SYNTHETASE BY HIGH-PERFORMANCE IMMUNOAFFINITY CHROMATOGRAPHY

被引：7

作者：

ALHAMA, J ^{[1
]}

LOPEZBAREA, J ^{[1
]}

TORIBIO, F ^{[1
]}

ROLDAN, JM ^{[1
]}

机构：

[1] UNIV CORDOBA, FAC VET, DEPT BIOQUIM & BIOL MOLEC, AVDA MED AZAHARA, E-14071 CORDOBA, SPAIN

来源：

JOURNAL OF CHROMATOGRAPHY | 1992年 / 589卷 / 1-2期

关键词：

D O I：

10.1016/0021-9673(92)80012-J

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

High-performance immunoaffinity chromatography (HPIAC) with anti-glutamine synthetase polyclonal antibodies bound to epoxy-activated silica was used to purify and determine this enzyme from the cyanobacterium Synechocystis. A single-step HPIAC procedure with cell-free extracts yielded electrophoretically homogeneous glutamine synthetase. In the determination of glutamine synthetase by HPIAC a linear response in the range 10-60-mu-g of enzyme was observed. Recoveries of 70% of the loaded enzymatic activity and 100% of protein were obtained. The determination of glutamine synthetase protein by HPIAC was compared with that obtained by rocket immunoelectrophoresis. The chromatographic method is proposed as a possible alternative to other immunochemical quantitative techniques, particularly when non-limiting amounts of samples are available.

引用

页码：121 / 126

页数：6

共 33 条

[1] HIGH-PERFORMANCE AFFINITY-CHROMATOGRAPHY OF NADP+ DEHYDROGENASES FROM CELL-FREE-EXTRACTS USING A NUCLEOTIDE ANALOG AS GENERAL LIGAND [J].

ALHAMA, J ;

LOPEZBAREA, J ;

TORIBIO, F .

JOURNAL OF CHROMATOGRAPHY, 1991, 586 (01) :51-59

[2] REGULATION OF CORN LEAF NITRATE REDUCTASE .1. IMMUNOCHEMICAL METHODS FOR ANALYSIS OF THE ENZYMES PROTEIN-COMPONENT [J].

CAMPBELL, WH ;

REMMLER, JL .

PLANT PHYSIOLOGY, 1986, 80 (02) :435-441

[3]

DEAN PDG, 1985, AFFINITY CHROMATOGR, P1

[4] PURIFICATION OF NITRATE REDUCTASE FROM SPINACH (SPINACEA-OLERACEA L) BY IMMUNOAFFINITY CHROMATOGRAPHY USING A MONOCLONAL-ANTIBODY [J].

FIDO, RJ .

PLANT SCIENCE, 1987, 50 (02) :111-115

[5]

FLORENCIO FJ, 1973, BIOCHIM BIOPHYS ACTA, V838, P233

[6] HPLC ANALYSIS OF HUMAN EPIDERMAL GROWTH-FACTOR USING IMMUNOAFFINITY PRECOLUMN .2. DETERMINATION OF HEGFS IN BIOLOGICAL-FLUIDS [J].

HAYASHI, T ;

SAKAMOTO, S ;

WADA, I ;

YOSHIDA, H .

CHROMATOGRAPHIA, 1989, 27 (11-12) :574-580

[7] NEW MEDIUM FOR ISOLATION AND GROWTH OF AUXOTROPHIC MUTANTS OF BLUE-GREEN-ALGA ANACYSTIS-NIDULANS [J].

HERDMAN, M ;

DELANEY, SF ;

CARR, NG .

JOURNAL OF GENERAL MICROBIOLOGY, 1973, 79 (DEC) :233-237

[8] HIGH-PRESSURE AFFINITY-CHROMATOGRAPHY OF CALMODULIN ON A PHENOTHIAZINE-SILICA [J].

JARRETT, HW .

JOURNAL OF CHROMATOGRAPHY, 1986, 363 (02) :456-461

[9]

KHURANA AL, 1989, J LIQ CHROMATOGR, V12, P3019

[10] CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].

LAEMMLI, UK .

NATURE, 1970, 227 (5259) :680-+

← 1 2 3 4 →