title: Development and validation of an indirect competitive ELISA for quantification of recombinant staphylokinase in rabbit plasma: Application to pharmacokinetic study.
abstract: The relatively short circulatory half-life (2-3 min) of staphylokinase is a major drawback in the development of SAK- (staphylokinase) based thrombolytic drug. A rapid and sensitive method, based on indirect competitive ELISA, was developed and validated for quantitative determination of SAK in rabbit plasma. The dynamic range of the assay varied between 0.41 ± 0.16 μg/L and 9.03 ± 0.38 μg/L (R(2) = 0.98) for SAK in rabbit plasma. There were no dilution linearity issues apparent with this assay. The precision (% CV) ranged from 4.6-9.7% for the intraassay and from 17.1-19.3% for interassay. This validated method was successfully employed for evaluation of various pharmacokinetic parameters of SAK in rabbit.
authors.affiliation: a Centre for Cellular & Molecular Biology , Hyderabad , India.|b Department of Biotechnology , Acharya Nagarjuna University , Guntur , India.
authors.forename: Anmol
authors.initials: A
authors.lastname: Kumar
authors.affiliation: c Centre for Bioseparation Technology , VIT University , Vellore , India.
authors.forename: K K
authors.initials: KK
authors.lastname: Pulicherla
authors.affiliation: b Department of Biotechnology , Acharya Nagarjuna University , Guntur , India.
authors.forename: K R S
authors.initials: KR
authors.lastname: Sambasiva Rao
mesh_terms: D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards*; D005260:Female; D005343:Fibrinolytic Agents / Q000097:blood / Q000493:pharmacokinetics*; D008297:Male; D008666:Metalloendopeptidases / Q000097:blood* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D011994:Recombinant Proteins / Q000097:blood / Q000378:metabolism
chemical_list: D005343:Fibrinolytic Agents; D011994:Recombinant Proteins; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus
keywords: half-life; immunoassay; indirect competitive ELISA; pharmacokinetics; staphylokinase; validation