PICOGRAM DETECTION LEVELS OF ASIALOFETUIN VIA THE CARBOHYDRATE MOIETIES USING THE LIGHT ADDRESSABLE POTENTIOMETRIC SENSOR

被引:11
作者
DILL, K
OLSON, JD
机构
[1] Molecular Devices Corporation, Sunnyvale, 94089, CA
关键词
ASIALOFETUIN; FETAL CALF SERUM; RICIN TOXIN; LIGHT ADDRESSABLE POTENTIOMETRIC SENSOR;
D O I
10.1007/BF00731262
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Fetal calf serum asialofetuin was assayed in the sandwich format using biotinylated and fluoresceinated ricin toxin (B-RCA and F-RCA). The sandwiched species was captured on a biotin-BSA coated nitrocellulose membrane with streptavidin. Anti-fluorescein antibody-urease conjugate was bound to the complex, and detected and quantitated under microvolume conditions using the light addressable potentiometric sensor. As little as 250 pg of asialofetuin was detectable whereas fetuin gave no response at conditions as high as 32 ng. Using a competitive inhibition assay, we established that the binding constant for the asialofetuin-ricin complex was 3.6 x 10(8) M(-1). This is in good agreement with data published using glycopeptides derived from asialofetuin, and RCA and the ricin agglutinin, RCA(120).
引用
收藏
页码:660 / 663
页数:4
相关论文
共 17 条
[2]   LECTIN-CARBOHYDRATE INTERACTIONS - STUDIES OF THE NATURE OF HYDROGEN-BONDING BETWEEN D-GALACTOSE AND CERTAIN D-GALACTOSE-SPECIFIC LECTINS, AND BETWEEN D-MANNOSE AND CONCANAVALIN-A [J].
BHATTACHARYYA, L ;
BREWER, CF .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1988, 176 (01) :207-212
[3]   NATURAL-ABUNDANCE, C-13-NUCLEAR MAGNETIC RESONANCE-SPECTRAL STUDIES OF CARBOHYDRATES LINKED TO AMINO-ACIDS AND PROTEINS [J].
DILL, K ;
BERMAN, E ;
PAVIA, AA .
ADVANCES IN CARBOHYDRATE CHEMISTRY AND BIOCHEMISTRY, 1985, 43 :1-49
[4]   ANTIBODY-ANTIGEN BINDING CONSTANTS DETERMINED IN SOLUTION-PHASE WITH THE THRESHOLD MEMBRANE-CAPTURE SYSTEM - BINDING CONSTANTS FOR ANTIFLUORESCEIN, ANTI-SAXITOXIN, AND ANTI-RICIN ANTIBODIES [J].
DILL, K ;
LIN, M ;
POTERAS, C ;
FRASER, C ;
HAFEMAN, DG ;
OWICKI, JC ;
OLSON, JD .
ANALYTICAL BIOCHEMISTRY, 1994, 217 (01) :128-138
[5]  
DILL K, 1993, 1993 P US ARM EDG RE
[6]  
DILL K, 1987, ADV CARBOHYD CHEM BI, V45, P170
[7]  
FRENOY JP, 1988, LECTINS BIOL BIOCH C, V6, P285
[8]  
GOTTSCHALK A, 1972, GLYCOPROTEINS THEIR, V5
[9]   LIGHT-ADDRESSABLE POTENTIOMETRIC SENSOR FOR BIOCHEMICAL SYSTEMS [J].
HAFEMAN, DG ;
PARCE, JW ;
MCCONNELL, HM .
SCIENCE, 1988, 240 (4856) :1182-1185
[10]  
MONTFORT W, 1987, J BIOL CHEM, V262, P5398