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RANIBIZUMAB ELISA (MAB-BASED)

RANIBIZUMAB ELISA (MAB-BASED)

Enzyme immunoassay for the specific and quantitative determination of free Ranibizumab (Lucentis®) in aqueous humour.
The solid phase (MTP) is coated by a highly specific monoclonal antibody directed against Ranibizumab. Therefore any cross reactivity to the other therapeutical monoclonal antibodies is excluded except for Bevacizumab.
 
Required Volume (µL)5
Incubation Time (min)135
Sampleaqueous humour
Plate Size96 Tests
Standard Range (ng/mL, 10x)0-300
Detection Limit (ng/mL)0,33
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.
 
Lucentis® is a trademark of ©2018 Genentech USA, Inc.

  • SPECIFICITY

    There is no cross reaction with any other proteins present in native aqueous humour samples. A screening test was performed with different aqueous humour samples. All produced OD450/620 nm values (ranged from 0.016 to 0.023) less than the mean OD (0.134) of standard D (1 ng/mL). In addition, binding of Ranibizumab to the solid phase is inhibited by recombinant human VEGF. Therefore, the ImmunoGuide Ranibizumab ELISA (mAb-Based) measures the biologically active free form of Ranibizumab, i.e. not pre-occupied by human VEGF antigen. No cross reaction was observed with the other therapeutic antibodies including Infliximab, Rituximab, Veolizumab, Trastuzumab, Nivolumab and Aflibercept at concentrations tested up to 50 µg/mL. All produced mean OD450/620 nm values (ranged from 0.052 to 0.068) less than standard D. Because the 2A1 mAb, used for coating solid phase, is also reactive against Bevacizumab, it causes full cross reaction. But a quantification of Bevacizumab in aqueous homour by this testkit is possible only by using the drug-specific standards (i.e. Bevacizumab).

     

    SENSITIVITY

    The lowest detectable level that can be clearly distinguished from the zero standard is 0,33 ng/mL (zero standard +2SD read from the curve) under the above-described conditions. If needed, the minimum measurable level of Ranibizumab in samples can be improved up to 10 times by diluting aqueous humour samples at 1:200 instead of 1:2000. Under the conditions described in section 10.2 the analytical sensitivity is 0,33 ng/mL, and corresponding to the detection limit (limit of quantification) of 0.66 µg/mL for undiluted clinical samples because the serum or plasma samples are instructed to be diluted at 1:2000 before starting the assay.

     

    PRECISION

    Intra-assay CV: <10%.

    Inter-assay CV: <10%.

     

    RECOVERY

    Recovery rate was found to be >90% with aqueous humour when spiked with exogenous Ranibizumab.

     

    AUTOMATION

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

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