CYCLIC-AMP RESPONSE TO RECOMBINANT HUMAN RELAXIN BY CULTURED HUMAN ENDOMETRIAL CELLS - A SPECIFIC AND HIGH THROUGHPUT INVITRO BIOASSAY

被引:86
作者
FEI, DTW [1 ]
GROSS, MC [1 ]
LOFGREN, JL [1 ]
MORAWORMS, M [1 ]
CHEN, AB [1 ]
机构
[1] GENENTECH INC,ASSAY SERV,SAN FRANCISCO,CA 94080
关键词
D O I
10.1016/0006-291X(90)91262-Q
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
A specific and high throughput 96-well format bioassay for recombinant human relaxin (rhRLX) has been developed using human endometrial cells (NHE cells). rhRLX caused a time- and dose-dependent stimulation of cyclic AMP (cAMP) with 1 2 maximal activity of 3.56±0.65 ng/ml (n=30). The range of the standard curve was 0.39 to 25 ng/ml with interplate precision of 17 and 22% CV for high and low controls respectively. The cAMP response requires forskolin and 3-isobutyl-1-methylxanthine, and is enhanced by prostaglandin E2 and F2α. The NHE cells do not respond to A or B chains of rhRLX, or a whole array of hormones. Preincubation of rhRLX with specific monoclonal antibody completely abolished the cAMP response. This bioassay has been used to determine the biological activity of several manufactured lots of recombinant human relaxin. © 1990.
引用
收藏
页码:214 / 222
页数:9
相关论文
共 22 条
[1]   RELAXIN-DEPENDENT ADENOSINE-3',5'-MONOPHOSPHATE CONCENTRATION CHANGES IN MOUSE PUBIC SYMPHYSIS [J].
BRADDON, SA .
ENDOCRINOLOGY, 1978, 102 (04) :1292-1299
[2]  
CASEY ML, 1984, IN VITRO CELL DEV B, V20, P396
[3]   THE EFFECT OF RELAXIN ON CYCLIC ADENOSINE 3', 5'-MONOPHOSPHATE CONCENTRATIONS IN HUMAN ENDOMETRIAL GLANDULAR EPITHELIAL-CELLS [J].
CHEN, G ;
HUANG, JR ;
TSENG, L .
BIOLOGY OF REPRODUCTION, 1988, 39 (03) :519-525
[4]   HUMAN RELAXIN INCREASES CYCLIC-AMP LEVELS IN CULTURED ANTERIOR-PITUITARY-CELLS [J].
CRONIN, MJ ;
MALASKA, T ;
BAKHIT, C .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1987, 148 (03) :1246-1251
[5]   THE PHARMACOKINETICS AND PHARMACODYNAMICS OF A HUMAN RELAXIN IN THE MOUSE PUBIC SYMPHYSIS BIOASSAY [J].
FERRAIOLO, BL ;
CRONIN, M ;
BAKHIT, C ;
ROTH, M ;
CHESTNUT, M ;
LYON, R .
ENDOCRINOLOGY, 1989, 125 (06) :2922-2926
[6]   THE EFFECT OF RELAXIN ON CYCLIC ADENOSINE-3',5'-MONOPHOSPHATE CONCENTRATIONS IN RAT MYOMETRIAL CELLS IN CULTURE [J].
HSU, CJ ;
MCCORMACK, SM ;
SANBORN, BM .
ENDOCRINOLOGY, 1985, 116 (05) :2029-2035
[7]   MODULATION OF CYCLIC-AMP IN ISOLATED RAT UTERINE TISSUE-SLICES BY PORCINE RELAXIN [J].
JUDSON, DG ;
PAY, S ;
BHOOLA, KD .
JOURNAL OF ENDOCRINOLOGY, 1980, 87 (01) :153-159
[8]  
KARIN S, 1971, ANN NY ACAD SCI, P483
[9]  
KRAMER SM, 1990, IN PRESS IN VITRO CE
[10]   THE EFFECTS OF ESTROGENS, PROGRESTAGENS, AND RELAXIN IN PREGNANT AND NONPREGNANT LABORATORY RODENTS [J].
KROC, RL ;
STEINETZ, BG ;
BEACH, VL .
ANNALS OF THE NEW YORK ACADEMY OF SCIENCES, 1959, 75 (02) :942-980