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Establishment of Homogeneous Time-resolved Fluorescence (HTRF) Method for Measurement of Monoclonal Antibody Concentration
DENG Chun-pin, QIN Yu-min, DENG Qiao-chun, WANG Bo, PENG Yu-cai
China Biotechnology ›› 2013, Vol. 33 ›› Issue (12) : 64-68.
PDF(665 KB)
PDF(665 KB)
Establishment of Homogeneous Time-resolved Fluorescence (HTRF) Method for Measurement of Monoclonal Antibody Concentration
A rapid and simple antibody analysis method based on HTRF technology was established to measure concentration of recombinant monoclonal antibody in both cell culture medium and purified antibody solution. APC-labeled antigen,antibody samples or controls, and Eu-labeled anti-human IgG are added into 96 well plate (Black), incubate for some time, followed by fluorescence reading with microplate reader. Standard curve was generated by 4-PL model, antibody concentration in tested samples was calculated accordingly by software. The method qualification results showed that the method is antigen-antibody specific, with accuracy close to 98.9%, precision close to 4.3%, and linearity range 25～1000 ng/ml. This method can be used for cell line screening, bio-process monitoring, and quality control during antibody drug development.
Homogeneous Time-resolved Fluorescence / FRET / Monoclonal antibody {{custom_keyword}} /
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