HIGH-AFFINITY RABBIT ANTIBODIES DIRECTED AGAINST METHOTREXATE - PRODUCTION, PURIFICATION, CHARACTERIZATION, AND PHARMACOKINETICS IN THE RAT

被引:15
作者
BALTHASAR, JP [1 ]
FUNG, HL [1 ]
机构
[1] SUNY BUFFALO,DEPT PHARMACEUT,BUFFALO,NY 14260
关键词
D O I
10.1002/jps.2600840103
中图分类号
R914 [药物化学];
学科分类号
100701 ;
摘要
We describe the production, purification, characterization, and disposition of rabbit polyclonal methotrexate antibodies. These antibodies are prepared for subsequent testing of a drug delivery approach to reduce systemic toxicity upon regional administration of methotrexate. The polyclonal antibodies were raised in New Zealand white female rabbits immunized with a methotrexate-bovine serum albumin conjugate. The anti-methotrexate antibodies were sequestered from rabbit serum through the use of a protein-G affinity column which allowed for purification of up to 100 mg of rabbit IgG in 30-40 min. The extent of purification was demonstrated through SDS-PAGE and calculation of specific binding activity relative to total protein concentration. The purified antibodies have been shown to have high affinity (K-eq, range: 1.8 x 10(8) to 8.75 x 10(9) M(-1)) and high selectivity for methotrexate. Preliminary pharmacokinetic studies of the purified antibodies in the rat following a 6 mg/kg intravenous infusion (n = 4) indicate a steady state volume of distribution of 38.0 +/- 11.2 mL kg(-1), a systemic clearance of 0.92 +/- 0.67 mL kg(-1) h(-1) and an elimination half life of 28.9 +/- 7.9 h (mean +/- SD).
引用
收藏
页码:2 / 6
页数:5
相关论文
共 27 条
[1]  
BALIS FM, 1989, AM J PEDIAT HEMATOL, V11, P74
[2]  
BALTHASAR J, 1994, J PHARMACOL EXP THER, V268, P734
[3]  
BREMNES RM, 1989, CANCER RES, V49, P2460
[4]   ANALYTICAL CONTROL PROCEDURES OF IMMUNOREACTIVITY FOR IGG AND FAB FRAGMENTS SPECIFIC TO HAPTENS [J].
CANO, NJ ;
SABOURAUD, AE ;
URTIZBEREA, M ;
CARCAGNE, J ;
GRANDGEORGE, M ;
SCHERRMANN, JMG .
PHARMACEUTICAL RESEARCH, 1992, 9 (05) :643-647
[5]   GRANULOCYTE-COLONY-STIMULATING FACTOR - EFFECTIVE IN AMELIORATING FLUOROURACIL-BASED MYELOSUPPRESSION [J].
CHRISTMAN, K ;
SALTZ, L ;
SCHWARTZ, G ;
FRIEDRICH, C ;
KELSEN, D .
JOURNAL OF THE NATIONAL CANCER INSTITUTE, 1993, 85 (10) :826-827
[6]  
CLAY EF, 1992, HEMATOL ONCOL CLIN N, V6, P915
[7]  
DEDRICK RL, 1978, CANCER CHEMOTHER PHA, V1, P161
[8]  
DONEHOWER RC, 1979, CLIN PHARMACOL THER, V26, P63
[9]  
Gibaldi M., 1982, PHARMACOKINETICS, V2nd ed., P409
[10]   LONG-DURATION INTRACAVITARY INFUSION OF METHOTREXATE WITH SYSTEMIC LEUCOVORIN PROTECTION IN PATIENTS WITH MALIGNANT EFFUSIONS [J].
HOWELL, SB ;
CHU, BBF ;
WUNG, WE ;
METHA, BM ;
MENDELSOHN, J .
JOURNAL OF CLINICAL INVESTIGATION, 1981, 67 (04) :1161-1170