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BEVACIZUMAB ELISA (MAB-BASED)

BEVACIZUMAB ELISA (MAB-BASED)

Enzyme immunoassay for the specific and quantitative determination of free Bevacizumab (Avastin) in serum and plasma.
The solid phase (MTP) is coated by a highly specific monoclonal antibody directed against Bevacizumab. Therefore any cross reactivity to the other therapeutical monoclonal antibodies, even with its 5 pointmutation Fab counterpart (Ranibizumab) is excluded.
 
Required Volume (µL) 10
Incubation Time (min) 100
Sample Serum or Plasma
Plate Size96 Tests
Standard Range (µg/mL, 1000x) 0-200
Detection Limit (ng/mL)5
Spike Recovery (%) >95%
Shelf Life (years)2
 
Intended Use: This kit has been developed for the measurement of drug levels in research and diagnostic uses.  It is suitable for Therapeutical Drug Monitoring (TDM) purposes.
 
Avastin® is a trademark of Genentech Inc./Roche.

  • SPECIFICITY

    There is no cross reaction with any other proteins present in native human serum. A screening test was performed with 48 different native human sera. All produced OD450/620 nm values (ranged from 0.022 to 0.042) less than the mean OD (0.132) of standard D (6 ng/mL). In addition, binding of Bevacizumab is inhibited by recombinant human VEGF-A in a concentration dependent manner. Therefore, the ImmunoGuide Bevacizumab ELISA (mAb-Based) measures the biologically active free form of Bevacizumab, i.e. not pre-occupied by VEGF. No cross reaction was observed with sera spiked with the other therapeutic antibodies including Infliximab, Adalimumab, Etanercept, Rituximab, Tocilizumab, Trastuzumab, Aflibercept and Golimumab at concentrations up to 2 mg/mL. All produced mean OD450/620 nm values ranged from 0.009 to 0.022. In addition, there is no cross reaction with Ranibizumab as well. Because when anti-human kappa monoclonal antibody was used as the conjugate instead, Ranibizumab did not bind to MAY-2B5 mAb-coated plate.

     

    SENSITIVITY

    The lowest detectable level that can be clearly distinguished from the zero standard is 2 ng/mL (zero standard +2SD read from the curve) under the above-described conditions. Analytical sensitivity is 2 ng/mL, and corresponding to the detection limit (limit of quantification) of 2 µg/mL for undiluted clinical samples because the serum or plasma samples are instructed to be diluted at 1:1000 before starting the assay.

     

    PRECISION

    Intra-assay CV: <10%.

    Inter-assay CV: <10%.

     

    RECOVERY

    Recovery rate was found to be >95% with native human serum and plasma samples when spiked with exogenous Bevacizumab at 200 µg/mL, 60 µg/mL, 20 µg/mL or 6 µg/mL.

     

    AUTOMATION

    The ImmunoGuide Bevacizumab ELISA (mAb-based) is suitable also for being used by an automated ELISA processor.

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