A sandwich ELISA for assessment of pharmacokinetics of HSA-(BNP)2 fusion protein in mouse plasma.
Brain natriuretic peptide (BNP) is a circulating hormone of cardiac origin that plays an important role in the regulation of intravascular blood volume and vascular tone. HSA-(BNP)(2), derived from the joining of human BNP to the C-terminus of human serum albumin (HSA), has been developed to prolong the BNP pharmacodynamic action. For the analysis of pharmacokinetics of the new drug, a novel sandwich enzyme-linked immunosorbent assay (ELISA) was established and validated to quantify HSA-(BNP)(2) fusion protein in mouse plasma. The ELISA method was calibrated with 1:10 and 1:100 dilutions of blank mouse plasma spiked with HSA-(BNP)(2) standard and validated with respect to parallelism, precision (intra- and inter-assay variation), accuracy (recovery), specificity and stability. The practical working range was estimated to be 31.2-2000ng/ml with the limit of detection was 7.8ng/ml. Recoveries ranged from 80.5 to 108.4%, while the intra- and inter-assay precisions were <2.73% and <4.32%, respectively. The terminal half-life of HSA-(BNP)(2) was 2.14h, which had extended more than 40 times compared to 3.1min half-life of BNP monomer in mouse.
Ding
Yue-Di
YD
Laboratory of Molecular Pharmacology, School of Medicine and Pharmaceutics, Jiangnan University, Wuxi 214122, PR China.
Lei
Jian-Yong
JY
Chen
Yun
Y
Jin
Jian
J
D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards; D005260:Female; D006801:Humans; D008297:Male; D051379:Mice; D020097:Natriuretic Peptide, Brain / Q000097:blood* / Q000493:pharmacokinetics* / Q000592:standards; D011485:Protein Binding / Q000502:physiology; D011897:Random Allocation; D011993:Recombinant Fusion Proteins / Q000097:blood* / Q000493:pharmacokinetics* / Q000592:standards; D012709:Serum Albumin / Q000032:analysis / Q000493:pharmacokinetics* / Q000592:standards
D011993:Recombinant Fusion Proteins; D012709:Serum Albumin; D020097:Natriuretic Peptide, Brain