Study on the ELISA Double-antibody Sandwich Method and Pharmacokinetics of Recombinant Human Calcineurin B Subunit in Rat Plasma
10.6039/j.issn.1001-0408.2016.25.04
- VernacularTitle:大鼠血浆中重组人钙调磷酸酶B亚基的ELISA双抗体夹心法测定及其药动学研究Δ
- Author:
Kesheng HAN
;
Xueli XIE
;
Shuhong TIAN
;
Guilan XING
;
Jiang FU
;
Jun XING
;
Chunhua LIN
;
Peng TAN
;
Jiping SHAO
- Publication Type:Journal Article
- Keywords:
rhCNB polypeptide;
ELISA-antibody sandwich method;
Rat;
Quantitative detection;
Pharmacokinetics
- From:
China Pharmacy
2016;27(25):3468-3470
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To establish a method for determining recombinant human calmodulin B subunit(rhCNB)in rat plas-ma,and study its pharmacokinetics characteristics. METHODS:ELISA double-antibody sandwich method was adopted. 1 μg/ml rhCNB monoclonal antibody mAb was wrapped,added to the to-be-test sample,rhCNB polyclonal antibody pAb(dilution ratio of 1∶5 000)and HRP-labeled conjugate of anti-IgG(dilution ratio of 1∶10 000)were added. Using tetramethylbenzidine for develop-ing,microplate reader was conducted in wavelength of 450 nm to determine the absorbance value(OD value)and plasma concen-tration of 6 rats after 2,15,30,60,120,240,480,720 min of iv 2.5 mg/kg rhCNB,and the pharmacokinetic parameters were calculated by BAPP 3.0 software. RESULTS:The linear range of rhCNB were 0.195-12.5 ng/ml(r2=0.995 0),lower limit of quan-titation was 0.195 ng/ml,accuracy were 97.300%-103.622%(RSD<7.5%,n=6);RSDs of within-batch,inter-batch,freezing and thawing 3 times were no higher than 8.5%(n=6,18,15). rhCNB pharmacokinetics characteristics in rat fitted to two-com-partment model,AUC0-720 min was 173.038 mg·min/L and t1/2 was 94.62 min. CONCLUSIONS:The established method has high specificity and sensitivity,good accuracy and precision,which can be used for rhCNB quantitative detection and pharmacokinetics study in biological samples.
