Open AccessAn enzyme immunoassay for determining plasma concentrations of didemnin B
Author(s) -
Raybould T. J. G.,
Grothaus P. G.,
Simpson Samantha B.,
Bignami G. S.,
Lazo Carolyn B.,
Newman R. A.
Publication year - 1992
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.1860060307
Subject(s) - immunoassay , enzyme , chemistry , plasma , chromatography , biochemistry , antibody , medicine , immunology , physics , quantum mechanics
Didemnin A was conjugated at the amino terminus of the N ‐methylleucine residue, via the linkers N ‐succinimidyl‐3‐(2‐pyridyldithio)propionate and trans‐1,4‐maleimidomethylcyclohexane carboxylic acid, to keyhole limpet hemocyanin (KLH) and bovine serum albumin (BSA). The didemnin‐KLH conjugates were used to hyperimmunize rabbits. The resulting high titer antisera were employed with didemnin‐BSA conjugate‐coated microtiter plate wells to develop an indirect competitive inhibition enzyme immunoassay (CIEIA) that was fully cross reactive with didemnin B. A CIEIA is described that is capable of detecting the drug in plasma from didemnin Btreated patients at concentrations down to 1‐3 ng/ml. This simple, sensitive CIEIA has been employed to demonstrate plasma drug clearance profiles with samples from didemnin B‐treated patients. © 1992 Wiley‐Liss, Inc.