Development of a competitive ELISA for the quantification of F5 conjugate in HER2-targeted STEALTH immunoliposome doxorubicin in plasma samples |
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Authors: | Ran Hu Rebecca Davis Yongjin Yao Yaodong Xu |
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Institution: | (1) Department of Bioanalysis, ALZA Corporation, 1900 Charleston Road, Mountain View, CA 94043, USA |
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Abstract: | HER2 (human epidermal growth factor receptor 2, erbB2, or neu) is overexpressed by a large number of tumor types and has been
identified as an important target for cancer therapy. F5 is a single-chain human antibody fragment that recognizes HER2 receptor
and is covalently conjugated to PEGylated lipid to form F5 conjugate (F5CG) in the product HER2 targeted STEALTH immunoliposome
doxorubicin. Here we described the method development of a competitive enzyme-linked immunosorbent assay (ELISA) for the determination
of total concentration of F5 conjugate in plasma samples. The method involved the biotinylation of F5CG, detergent treatment
of plasma sample to solubilize F5CG into monomeric form, and competitive ELISA for solubilized F5CG competitively binding
to anti-F5CG antibody with biotinylated F5CG for the determination of total F5CG in plasma. The detection range of this method
was from 0.2 ng/mL to 125 ng/mL for F5CG in plasma. The lower limit of quantification (LLOQ) was 0.2 ng/mL. This method was
established and used for the measurement of F5CG concentration to provide information about F5CG circulation after the administration
of immunoliposome in preclinical studies.
![MediaObjects/216_2006_733_Figa_HTML.gif](/content/83377141p717u25p/MediaObjects/216_2006_733_Figa_HTML.gif) |
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Keywords: | Competitive ELISA Assay development F5 conjugate (F5CG) Targeted immunoliposomal formulation HER2 receptor |
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